ポスター
F. 神経系の疾患 2
F. Disorders of the Nervous System 2 |
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| 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-097
糖尿病性神経障害モデルショウジョウバエにおけるグリア細胞プロテアソームの役割
Role of glial proteasome in a Drosophila model of diabetic peripheral neuropathy
*鈴木 マリ(1)、黒見 坦(2)、進藤 真由美(3)、新見 直子(1)、齊藤 実(2)、三五 一憲(1)
1. 東京都医学総合研究所 糖尿病性神経障害プロジェクト、2. 東京都医学総合研究所 学習記憶プロジェクト、3. 東京都医学総合研究所 基盤技術支援センター
*Mari Suzuki(1), Hiroshi Kuromi(2), Mayumi Shindo(3), Naoko Niimi(1), Minoru Saitoe(2), Kazunori Sango(1)
1. Diabetic Neuropathy Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan, 2. Learning and memory Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan, 3. Center for Basic Technology Research, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
Keyword: Diabetic peripheral neuropathy, high-sugar diet, nociception, proteasome
Diabetic peripheral neuropathy (DPN) is the most common chronic and progressive complication of diabetes mellitus (DM). DPN causes chronic pain and paresthesias, but the most common symptom is sensory loss, which is a dominant risk factor for foot ulceration that leads to infections and foot amputations. In this study, we used Drosophila melanogaster, which is suitable for genetic analyses, for developing a dietary-induced DPN model to explore novel genes that modulate DPN-like phenotypes. Feeding adult wild-type flies a high-sugar diet (HSD) induced metabolic dysfunction, including hyperglycemia and insulin resistance. We tested whether HSD-fed flies exhibit altered response to heat stimuli by using the heat avoidance (HA) test, and found that the avoidance of noxious heat was impaired upon HSD feeding, suggesting that sensory loss was induced. The HA impairment was associated with a shrinkage of leg neurons expressing the Drosophila TRP channel Painless, which is essential for thermal nociception. To identify genes that modify the HA impairment of HSD-fed flies, we performed genetic screening and found that systemic knockdown of the proteasome modulator 9 (PSMD9) suppressed the HA impairment. The HA impairment was also suppressed by an administration of proteasome inhibitor Ixazomib. In addition, glia-specific knockdown of the PSMD9 or proteasomal subunit (Rpt4 or Rpt5), and glia-specific expression of the dominant- negative mutant proteasome catalytic subunit β2 and β6 suppressed the HA impairment, suggesting that suppression of proteasomal activity in glia plays a protective role in the HA impairment induced by HSD. Proteomics analysis revealed that Ixazomib induces a remarkable elevation of heat-shock proteins in cultured glial cells, and glial knockdown of Drosophila DNAJ1, one of the HSP40 family, abolished the effect of Ixazomib on HSD-induced HA impairment. These results demonstrate that glial proteasome activity and heat-shock proteins can regulate HA impairment induced by HSD in adult Drosophila. Our findings propose that the glial protein homeostasis system is one of the therapeutic targets for the DPN. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-098
気圧低下がSpreading Depressionを生じるか？～低気圧に伴う片頭痛発作のメカニズム
Does a decrease in atmospheric pressure cause Spreading Depression? - Mechanism of migraine attacks associated with low pressure
*前田 仁士(1)、秋本 悠香(1)、桑名 俊一(1)
1. 植草学園大学
*Hitoshi Maeda(1), Yuuka Akimoto(1), Shunichi Kuwana(1)
1. Uekusagakuen University
Keyword: migraine, spreading depression, imaging
Migraine is a severe and disabling condition and a public health problem which impacts on not only the patient and its family but society. One of the causes of migraine is said to change in atmospheric pressure or temperature due to climate changes, and there are reports that it has been confirmed that several hPa changes in atmospheric pressure can cause headache attacks. There are also reports of exacerbation of chronic pain when a decrease in atmospheric pressure within a change in the weather, occurred in the specific chamber, which can change atmospheric pressure in animal experiments.
Cortical spreading depression (CSD), a wave of cellular depolarization that propagates slowly across the brain surface followed by suppression of brain activity, has been presumed to be the physiological substrate of the migraine aura. However, its generation source has not been obvious yet. Therefore, we examined whether CSD can occur when this atmospheric pressure changes, using the autofluorescence of mitochondrial flavoproteins, which was presented previously at this conference: when blue (~465 nm) LED light was exposed, the intrinsic flavoprotein fluorescent signal changes were observed with the CCD camera system. We made a specific chamber attached a vacuum valve, which can artificially reduce internal pressure, and directly imaged cortical activity of the mouse with the scalp peeled in it. When the pressure in chamber began to decrease, there was an enhancement change in fluorescence from the tissue around the cerebrovascular vessel, and it was found that the fluorescence intensity increased by a barometric pressure drop of about 5 hPa, and it spread diffusely to neighboring tissues. The fluorescence of the blood vessels decreased in reverse. When the barometric pressure drop was stopped, the fluorescence changes also returned to normal. It was reported previously that chronic migraine model mice created by frequent administration of NTG are prone to CSD, but the fluorescence change in the brain surface by this experiment is not clear from that of control mice. The ease of diffusion was either different depending on each individual.
The propagation of the fluorescence changes observed was similar in response to the CSD reported last time, and it was possible to induce CSD by ordinary electrical stimulation after the barometric pressure stimulation experiment, so it was considered to be a reaction equivalent to CSD. Therefore, environmental changes such as atmospheric pressure changes can cause CSD, which can be a factor in migraine attacks. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-099
エーラス･ダンロス症候群テネイシンX欠損マウスにおける有髄A線維の応答過敏を介したアロディニア発症
Hypersensitivity of myelinated A fibers promotes mechanical allodynia in tenascin-X-deficient mice associated with Ehlers-Danlos syndrome
*芦高 恵美子(1)、鎌田 浩輝(1)、宇戸 禎仁(1)、川原 幸一(1)、南 敏明(2)、伊藤 誠二(2)、松本 健一(3)
1. 大阪工業大学大学院工学研究科化学・環境･生命工学専攻、2. 大阪医科薬科大学医学部麻酔科、3. 島根大学総合科学研究支援センター生体情報･RI
*Emiko Okuda-Ashitaka(1), Hiroki Kamada(1), Sadahito Uto(1), Koichi Kawahara(1), Toshiaki Minami(2), Seiji Ito(2), Ken-ichi Matsumoto(3)
1. Dept of Biomedical Engineering, Osaka Inst of Technology, Osaka, Japan, 2. Dept of Anesthesiology, Osaka Med Pharma Univ, Takatsuki, Japan, 3. Dept of Biosign and Radioiso Exp, Interdiscip Cent for Sci Res, Org for Res Acad Info, Shimane Univ, Izumo, Japan
Keyword: Ehlers-Danlos syndrome, Tenascin-X, Allodynia, A-fiber
Ehlers-Danlos syndrome is a human heritable disorder characterized by skin hyperextensibility, joint hypermobility, and easy bruising. Tenascin-X (TNX) is a member of the extracellular matrix glycoprotein tenascin family, and TNX deficiency leads to Ehlers-Danlos syndrome. TNX-deficient Ehlers-Danlos syndrome patients complain of chronic joint pain, myalgia, paresthesia, and axonal polyneuropathy. We previously have been reported that TNX-deficient mice exhibit mechanical allodynia and hypersensitivity of myelinated A-fibers (Sci Rep 10:6569, 2020). However, the hypersensitivity of myelinated A-fibers by which TNX deficiency complicates the mechanical allodynia are unknown. In the present study, we investigated pain response of TNX-deficient mice using pharmacological silencing of A fibers with co-injection of QX-314, a membrane-impermeable lidocaine analog, plus flagellin, a Toll-like receptor 5 ligand (TLR5). Intraplantar co-injection of QX-314 plus flagellin significantly inhibited the mechanical allodynia in TNX-deficient mice compared to the vehicle administration. Co-injection of QX-314 plus flagellin significantly reduced the hypersensitivity to transcutaneous sine wave stimuli at frequencies of 5 Hz (C fiber responses), 250 Hz (Ad fiber responses) and 2000 Hz (Aβ fiber responses) in TNX-deficient mice, although they reduced stimuli at frequency of 250 Hz and 2000 Hz, but not 5 Hz, in wild-type mice. QX-314 alone also inhibited the mechanical allodynia in TNX-deficient mice compared to the vehicle administration, and it reduced the hypersensitivity to transcutaneous sine wave stimuli at frequencies of 250 Hz and 2000 Hz, but not to stimuli at frequency of 5 Hz in TNX-deficient mice. These results suggest that the TNX deficiency-induced mechanical allodynia is mediated by A-fiber sensitivity. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-100
Neural correlates of pareidolic misperceptions in Parkinson’s disease patients
*Revankar Gajanan(1)、Kajiyama Yuta (1)、Hattori Noriaki(1,2)、Nakano Tomohito(1)、Shimokawa Tetsuya (3)、Mihara Masahito(1,4)、Mori Etsuro(1)、Mochizuki Hideki(1)
*Gajanan Revankar(1), Yuta Kajiyama(1), Noriaki Hattori(1,2), Tomohito Nakano(1), Tetsuya Shimokawa(3), Masahito Mihara(1,4), Etsuro Mori(1), Hideki Mochizuki(1)
1. Grad. School of Med, Osaka Uni, Osaka, Japan, 2. Faculty of Medicine, Univ of Toyama, Toyama, Japan, 3. CiNet, NICT, Osaka, Japan, 4. Kawasaki Med Sch, Kawasaki, Okayama, Japan
Keyword: Parkinson's disease, Visual hallucinations , EEG, Eye-tracking
Background: Parkinson’s disease (PD) patients susceptible to visual hallucinations encounter perceptual deficits in the form of pareidolias. Pareidolias are visual phenomena in which ambiguous forms or shapes appear meaningful due to inaccurate perception and serve as early markers for hallucinations.
Objective: We present here brain and behavioral correlates of pareidolias using a novel, multimodal, testing paradigm to reveal cortical fronto-parietal communication influencing top-down visual processing in PD patients.
Methods: 21 PD patients and 12 healthy controls were evaluated. Tests for cognition, attention and executive functions were performed. Pareidolias were evaluated using a computerized pareidolia test with synchronized EEG and eye-tracking. PD patients were divided into those with pareidolias (N=10) and those without (N=11). Responses were measured ‘during’ and ‘preceding’ the presentation of the stimuli. Fixation and saccade related parameters were used to assess eye movement behavior. EEG amplitudes were analyzed in the presaccadic phase. Finally, pre-stimulus EEG connectivity were evaluated using graph networks in the spectral domain (theta, low-alpha, and high-alpha frequencies).
Results: PD patients with high pareidolias were older and under-performed on neuropsychological tests. Eye-tracking data showed longer visit (saccade-dependent) and fixation durations for pareidolic stimuli ‘during’ active search task. Eye-tracking synched EEG showed higher presaccadic potentials on frontal electrodes independent of saccade sizes suggesting a stronger frontal activation. Pre-stimulus EEG low-alpha band networks showed a tendency towards higher frontal activity, increased normalized clustering coefficient with lower frontal degree centrality in PD with pareidolia group.
Conclusion: We found that evoking pareidolias specifically defines visuo-perceptual inadequacies in PD patients despite their wide range of cognitive scores. We suggest pareidolias in PD are due to an abnormal top-down modulation of visual processing which are defined by frontal cortex alterations that affects visual attention and guidance to ambiguous stimuli due to a dissonance between patients' internally generated mental processing with external stimuli. Our results highlight the importance of the use of pareidolia as a screening tool for neurodegenerative diseases, as well as implications for interventions in the future. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-101
Comparison of Cognitive Performance between Patients with Parkinson’s Disease and Dystonia Using an Intraoperative Recognition Memory Test
*Shi Lin(1)、Qin Guofan(1)、Xu Yichen(1)
*Lin Shi(1), Guofan Qin(1), Yichen Xu(1)
1. Beijing Tiantan Hospital, Capital Medical University, Beijing, China
Keyword: Intraoperative recognition memory test, Parkinson’s disease, dystonia, cognitive impairment
Neuroscientific studies on the function of the basal ganglia often examine the behavioral performance of patients with movement disorders, such as Parkinson’s disease (PD) and dystonia (DT), while simultaneously examining the underlying electrophysiological activity during deep brain stimulation surgery. Nevertheless, to date, there have been no studies comparing the cognitive performance of PD and DT patients during surgery. In this study, we assessed the memory function of PD and DT patients with the Montreal Cognitive Assessment (MoCA) and the Mini-Mental State Examination (MMSE). We also tested their cognitive performance during the deep brain stimulation surgery using a continuous recognition memory test. We compared the cognitive assessment and intraoperative test results between the PD and DT patients, and correlated the intraoperative cognitive performance to their preoperative cognitive assessment results. The results of the MoCA and MMSE failed to reveal significant differences between the PD and DT patients. Additionally, no significant difference was detected by the intraoperative memory test between the PD and DT patients. The intraoperative memory test scores were highly correlated with the MMSE scores and MoCA scores. Our data suggest that DT patients perform similarly to PD patients in cognitive tests during deep brain stimulation surgery, which provides potential possibilities to compare the brain activity between PD and DT patients and correlates with cognitive behaviours, and intraoperative memory tests can be used as a quick memory assessment tool during surgery. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-102
ニーマンピック病C型における免疫系による神経変性制御
Modulation of neurodegeneration by peripheral immune system in Niemann-Pick type C
*安田 徹(1)、内山 徹(1)、望月 秀樹(2)、小野寺 雅史(1)
1. 国立成育医療研究センター、2. 大阪大学大学院医学系研究科神経内科学
*Toru Yasuda(1), Toru Uchiyama(1), Hideki Mochizuki(2), Masafumi Onodera(1)
1. National Center for Child Health and Development, Tokyo, Japan, 2. Department of Neurology, Graduate School of Medicine, Osaka University, Osaka, Japan.
Keyword: Lysosome storage disrder, Niemann-Pick disease type C, Purkinje cell, immune system
Niemann-Pick disease type C (NPC) is an autosomal-recessively inherited lysosomal storage disorder affecting an estimated 1 in 120,000 live births worldwide. Mutations in NPC1 or NPC2 gene represent approximately 95% or 5% of total patients with NPC, respectively. Their gene products, NPC1 and NPC2 proteins, function cooperatively in late endosomes and lysosomes to transport unesterifed cholesterol to the plasma membranes and the other organelles in the cell. Typical clinical feature of the disease is neurovisceral accumulation of unesterified cholesterol and several forms of glycosphingolipids. NPC can present with a broad range of clinical manifestation from a neonatal acute fatality to an adult-onset chronic disease associated with neurodegeneration. The development of neurological symptoms, including cerebellar ataxia, laughter-induced cataplexy, dystonia, and progressive dementia, affects quality of life of the patients drastically. Hence, it is essential to explore the pathogenic events that trigger and/or promote the neurodegenerative process for future clinical interventions. In this study, we addressed an involvement of immune system in neuropathogenic process using a murine model of NPC. Breaching of blood-brain barrier and infiltration of monocyte-derived macrophages correlated spatially and temporally with the loss of cerebellar Purkinje cells, which is a hallmark of neurodegeneration in NPC. Reduction of microglial cells and circulating monocytes ameliorated Purkinje cell degeneration. On the other hand, lack of acquired immune system enhanced cerebellar ataxic phenotype and Purkinje cell loss, suggesting that lymphoid cells may have therapeutic effect in NPC. We found that peripheral injection of CD4/CD25-double-positive regulatory T cells ameliorated cerebellar ataxia and Purkinje cell loss. Our results disclose a previously unrecognized neuropathogenicity of immune system in NPC and would benefit future remedies for devastating neurological diseases. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-103
ASIC2a 点変異トランスジェニックラットの解析
Generation and phenotype of ASIC2a transgenic rat with gain-of-function point mutation
*柴田 泰宏(1)、熊本 奈都子(1)、佐久間 英輔(2)、石田 雄介(3)、植田 高史(1)、島田 昌一(4)、鵜川 眞也(1)
1. 名古屋市立大学大学院医学研究科機能組織学、2. 名古屋市立大学大学院医学研究科統合解剖学、3. 東北医科薬科大学医学部医学科組織解剖学、4. 大阪大学大学院医学系研究科神経細胞生物学
*Yasuhiro Shibata(1), Natsuko Kumamoto(1), Eisuke Sakuma(2), Yusuke Ishida(3), Takashi Ueda(1), Shoichi Shimada(4), Sinya Ugawa(1)
1. Dept ofAnatomy and Neuroscience, Grad Sch Med, Nagoya City Univ, Nagoya, Japan, 2. Dept of Integrative Anatomy, Grad Sch Med, Nagoya City Univ, Nagoya, Japan, 3. Div of Histology and Anatomy, Dept of Med, Tohoku Med and Pham Univ, Miyagi, Japan, 4. Dept of Neuroscience and Cell Biology, Grad sch Med, Osaka Univ, Osaka, Japan
Keyword: gain-of-function, ASIC2a, cerebellum
The acid-sensing ion channels (ASICs) are neuronal cation channels activated by extracellular protons, forming a superfamily composed of six major members. This superfamily is homologous of degenerin which is a mechanically-gated ion channel of C.elegans. In previous studies, specific amino acid substitutions in degenerin lead to gain-of-function mutation which convert the channels into constitutively active to induce the degeneration of neurons where they are expressed. ASIC2a is a member of the ASIC superfamily and is predominantly expressed in central neurons, and its gain-of-function mutation can stay open as the degenerin mutants are introduced into its gene. However, the phenotypes and physiological significance of mammals with such gain-of-function mutations in ASIC2a remains unclear. Here we generated heterozygous transgenic (Tg) rats expressing ASIC2a-G430F, which is the most active form of the gain-of-function mutants, under the control of the intrinsic ASIC2a promoter. The following considerations were scrutinized to clarify the details of the function, such as appearance observation, body weight measurement, rotarod test, footprint test, and histological examination of the brain were performed. The genome walking method was used to determine the transgene insertion site. We also identified the expression of PCP-2 as a Purkinje cell marker and GABA-A alpha 6 as a granule cell marker in the ASIC2a positive neuronal cells separated from the cerebellum by single-cell RT-PCR. To our knowledge, this is the first description of an ASIC gain-of-function mutant in mammals. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-104
マウス二次運動野における交連線維の光遺伝学的興奮によって誘発されるフィールド電位とスパイク発火特性への薬理学的影響
Pharmacological effect on characteristics of field potential and spike activity evoked by optogenetic callosal excitation in mouse secondary motor cortex
*半田 高史(1)、相澤 秀紀(1)
1. 広島大学大学院医系科学研究科（医）
*Takashi Handa(1), Hidenori Aizawa(1)
1. Grad Sch Biomed Health Sci (Med), Hiroshima Univ
Keyword: corpus callosum, stroke, neuromodulator
Interhemispheric connections through the corpus callosum allow interaction of the homotopic cortical regions between hemispheres. Indeed, the neuronal excitation in the mouse cerebral cortex leads to the transient inhibition of the homotopic cortical area on the contralateral side as known as the interhemispheric inhibition. However, it remains unclear how it is modulated in the pathological and recovering brain. Here, we addressed these questions through in vivo electrophysiological recording of local field potential and multiunit activity together with pharmacological and optogenetic manipulations in mice. We recorded neural activity in the secondary motor cortex (M2) of the left hemisphere in response to the neural excitation in the other M2 where neurons expressing channelrhodopsin-2. At the recording site, we topically applied agonists or antagonists of GABAB or cholinergic receptors. After a single-pulse optical stimulation (1 ms duration) over M2 of the right hemisphere, some neurons exhibited a brief increase of firing rate followed by inhibition of spiking for about a couple of hundreds millisecond. This firing pattern was observed while evoked field potentials occurred. To investigate the dynamics of synaptic potentials underlying this firing pattern, we analysed the characteristics of evoked field potential. The optical stimulation over M2 of the right hemisphere evoked response of field potential in M2 of the left hemisphere. We found that the early negative component of evoked field potential after optical stimulation was amplified by application of GABAB receptor antagonist (phaclofen), but not by application of cholinergic receptor agonist (carbachol) and antagonist (atropine). In contrast, the late positive component of evoked field potential was enlarged by cholinergic antagonist, but not by GABAB receptor drugs. Those results suggest that GABAB receptor and cholinergic receptor differentially influence the interhemispheric inhibition. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-105
頚椎症性脊髄症に対するニクジュヨウエキスの効果
Effects of Cistanche Tubulosa extract on cervical spondylotic myelopathy
*須山 真聡(1)、楊 熙蒙(1)、東田 千尋(1)
1. 富山大学 和漢医薬学総合研究所 神経機能学領域
*Masato Suyama(1), Ximeng Yang(1), Chihiro Thoda(1)
1. Section of Neuromedical Science, Institute of Natural Medicine, University of Toyama
Keyword: spinal cord
頚椎症性脊髄症 (Cervical spondylotic myelopathy : CSM)は、頚椎脊柱管が狭い状態での頚椎の加齢性変化による脊髄圧迫によって脊髄麻痺 (四肢のしびれ感、手足の巧緻運動障害、歩行障害、膀胱障害)を発症する疾患の総称である。頚椎の圧迫が高度の場合は手術適応となり、薬物療法としては鎮痛剤などの対症療法しかない。手術で圧迫を除去しても、それまでに進行した神経細胞のダメージによる機能障害は回復せず、根本的治療となるような機能改善薬の開発が望まれている。CSM患者での研究では、重症度に応じて軸索容量が減少することが示されており、臨床症状である運動機能や感覚機能の低下は脊髄中を走行する軸索の機能不全によってもたらされていると考えられる。しかし、CSMの治療研究として「変性した軸索を修復する治療法」が試された例は、ヒトにおいても動物においても未だない。そこで本研究は、軸索修復によるCSMの機能改善を目指した。候補薬物として、これまでの我々の研究において、脊髄損傷慢性期からの投与で運動機能を回復させたニクジュヨウエキスを検討した。 まず我々は、CSMの機能回復に関わる組織として、脊髄、脳、骨格筋を想定した。ニクジュヨウエキス経口投与後に、活性成分がそれらの組織に移行するかを調べた。以前の慢性期脊髄損傷の研究では、ニクジュヨウエキス中の活性成分はacteosideである結果を得ている。また、ニクジュヨウエキス中の主要成分としてacteosideのほかにechinacosideも相当量含有され、経口投与後のechinacosideは代謝酵素によりglucoseが外れてacteosideに変換されると一般的に考えられている。そこで、ニクジュヨウエキス経口投与0.5、１、3、24時間後に各組織を摘出し、その中のacteosideおよびechinacoside量をLC-MSを用いて測定した。その結果、acteosideのみならずechinacosideそのものも大脳皮質、脊髄および前脛骨筋へ移行した。そこでacteosideとechinacosideの神経細胞および骨格筋細胞への作用について検討した。胎生14日齢ddYマウスから大脳皮質神経細胞を初代培養しacteosideを4日間処置すると軸索が有意に伸展した。また、生後2日のddYマウスから初代培養した骨格筋細胞にacteosideを2日間処置し、その上に胎生14日齢ddYマウスの脊髄神経細胞を重層培養するとその軸索が有意に伸展した。いずれの実験でもechinacosideは無効だった。 以上、ニクジュヨウエキスの経口投与により中枢神経系と骨格筋に移行する成分のうち、acteosideには神経細胞に直接作用して軸索伸展を促進させる作用と、骨格筋に作用することで間接的に軸索伸展を促進させる作用はあることが示された。 これまでに報告されてきたいくつかのCSM動物モデルでは、機能障害と軸索障害の関係性が検討されておらず、また発症までに長期間を有する等の課題があった、そこで我々は新しいCSMモデルマウスを作製し、ニクジュヨウエキスの効果について検討している。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-106
テタヌストキシン側頭葉てんかんモデルにおける、細胞外グルコース濃度とてんかん性活動との関連について
Relationships between extracellular glucose concentration and epileptic activities in an epileptic animal model using tetanus toxin.
*中谷 光良(1)、Ghestem Antoine(2)、Ivanov Anton(2)、Bernard Christophe(2)
1. 順天堂大学、2. Aix Marseille University
*Mitsuyoshi Nakatani(1), Antoine Ghestem(2), Anton Ivanov(2), Christophe Bernard(2)
1. Juntendo University, 2. Aix Marseille University
Keyword: Animal epilepsy model, Interictal epileptic discharges, Ictal baseline shift, Glucose
[Objectives] Among the proposed seizure classifications based on seizure dynamics (Viktor et al., 2014), the saddle node (SN) onset, which is characterized by ictal baseline shift (IBS), seems to be predominant across species and was particularly observed in the early stage of the epileptogenicity. To test our hypotheses that metabolism can affect the emergence of this predominant SN onset activities, we aimed to investigate how glucose, which is fundamental energy source for neuron and glial cell, affect the occurrence patterns of interictal epileptic discharges (IEDs) and epileptic seizures using an epileptic animal model.[Methods] We enrolled 15 male Sprague-Dawley rats and 11 rats were treated with tetanus toxin (TeNT) to develop epileptic activities and spontaneous focal epileptic seizures. An insulated stainless electrode (for all) and glucose sensor (only for 7 rats) were simultaneously inserted after the injection of 30-50ng TeNT in CA3 region of ipsilateral hippocampus. We evaluated the occurrence pattern of IEDs, epileptic seizures and corresponding change of extracellular glucose level. Additionally, we evaluate phase locking value (PLV) between low and high frequency activities to see the characteristic electrophysiological change before SN onset activities.[Results] Six out of 11 rats treated with TeNT developed IEDs and spontaneous epileptic seizures. Interestingly, we found that aside from SN onset activities at seizure onset, baseline shift activities without subsequent epileptic discharges were observed during peri-ictal periods. Increase of PLV between delta and ripple range was observed before SN onset seizures, but not observed before peri-ictal baseline shift activities. The analysis in three rats who developed epileptic seizures with glucose sensor showed that the peri-ictal baseline shift activities required a relatively large amount of glucose compared to the same amplitude of IEDs and IBS.[Conclusions] Baseline shift activities may play a crucial role to activate neuron and glial cells by utilizing higher amounts of glucose possibly for preventing epileptic seizures. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-107
シンタキシン1Bのハプロ不全による熱性けいれんの発症機序の解明
Molecular mechanisms of fever-associated epilepsy syndrome caused by haploinsufficiency of syntaxin 1B
*三嶋 竜弥(1)、小藤 剛史(2)、藤原 智徳(1,3)、齋藤 綾子(1)、寺尾 安生(1)
1. 杏林大学医学部病態生理学教室、2. 杏林大学医学部RI部門、3. 埼玉医科大学保健医療学部臨床検査学科
*Tatsuya Mishima(1), Takefumi Kofuji(2), Tomonori Fujiwara(1,3), Ayako Saito(1), Yasuo Terao(1)
1. Dept Med Physiol, Kyorin Univ Sch Med, Tokyo, Japan, 2. RI Lab, Kyorin Univ Sch Med, Tokyo, Japan, 3. Sch Medical Technology, Fac Health & Medical Care, Saitama Med Univ, Saitama, Japan
Keyword: Syntaxin1B, febrile seizures, GABA
Two syntaxin 1 isoforms, HPC-1/Syntaxin 1A and Syntaxin 1B (STX1B), are coexpressed in neurons and function as neuronal target membrane (t)-SNAREs. We previously reported that STX1B is primarily involved in the regulation of different types of fast synaptic vesicle exocytosis, including spontaneous and evoked release of glutamatergic and GABAergic synaptic transmission. Recently, it has been reported that de novo heterozygous mutations in the STX1B gene, cause a familial fever-associated epilepsy syndrome. STX1B is an essential component of the presynaptic neurotransmitter release machinery as a SNARE protein that regulates the exocytosis of synaptic vesicles. It is also involved in regulating the functions of the SLC6 family of neurotransmitter transporters that reuptake neurotransmitters, including inhibitory neurotransmitters, such as GABA and glycine. The purpose of the present study was to elucidate the molecular mechanisms underlying the development of febrile seizures by examining the effects of STX1B haploinsufficiency on inhibitory synaptic transmission during hyperthermia in a mouse model. STX1B heterozygous (STX1B+/−) mice showed increased susceptibility to febrile seizures and drug-induced seizures. In cultured hippocampal neurons, we examined the temperature-dependent properties of neurotransmitter release and reuptake by GABA transporter-1 (GAT-1) at GABAergic neurons using whole-cell patch-clamp recordings. The rate of spontaneous quantal GABA release was reduced in STX1B+/− mice. The hyperthermic temperature increased the tonic GABAA current in wild-type (WT) synapses, but not in STX1B+/− synapses. In WT neurons, spontaneous recurrent bursting activities were reduced in a GABA-dependent manner at hyperthermic temperature; however, this was abolished in STX1B+/− neurons. The blockade of GAT-1 increased the tonic GABAA current and suppressed recurrent bursting activities in STX1B+/− neurons at the hyperthermic temperature. These data suggest that functional abnormalities associated with GABA release and reuptake in the presynaptic terminals of GABAergic neurons may increase the excitability of the neural circuit with hyperthermia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-108
Role of Increasing GLT-1 Expression in Repairing Cognitive and Neuronal Changes in a Rat Model of Epilepsy
*Ying-Jui Ho(1), Yu-Shiuan Tzeng(1), Wen-Yuan Liu(1), Jin-Yi Yao(1), Yun-Ju Hsieh(1), Yong-Zhu Zhuang(1)
1. Department of Psychology, Chung Shan Medical University, Taiwan
Keyword: epilepsy, astrocyte, glutamate, GLT-1
The pathophysiology of epilepsy includes the imbalance of neurotransmitter levels, which is caused by excessive level of excitatory neurotransmitter, glutamate, and insufficient level of inhibitory neurotransmitter, GABA. Excessive neuronal excitation not only results in seizure and neuronal loss but also hyper-activation in the subthalamic nucleus (STN). Astrocytes uptake extracellular glutamate via glutamate transporter-1 (GLT-1), thereby decreasing neuronal excitation. In addition, astrocytes can convert the uptaken glutamate into glutamine that is a precursor of GABA synthesis, thereby increasing neuronal inhibition. Therefore, astrocytes may be an effective treatment target of epilepsy. Repeated seizures not only damage GABAergic neurons, decrease GLT-1 expression but also impair astrocytic function. SUL, one of β-lactam drug, has neuroprotection potential by up-regulating GLT-1 expression. The purpose of this study was to evaluate whether SUL have neuroprotection by increasing GLT-1 expression on astrocytes in an epilepsy rat model. We used pentylenetetrazol to induce epilepsy kindling rat model and histological analysis to evaluate neuronal changes in the rats. We measured neuronal density by Nissl staining, analyzed astrocytic GLT-1 expression by immunofluorescence, and detected neuronal activity by cytochrome oxidase staining. The results showed that SUL treatment restored neuronal density and astrocytic GLT-1 expression in the hippocampus of epileptic rats. Additionally, SUL suppressed seizure related hyperactivity of STN. In conclusion, SUL shows neuroprotection by increasing GLT-1 expression on astrocyte in epilepsy rat model. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-109
発作間欠期における多脳領域間オシレーションパターンによる発作感受性の解読および定量化
Decoding and quantification of seizure susceptibility with cross-regional oscillatory brain activity patterns in a rat model of temporal lobe epilepsy
*竹内 雄一(1,2)、李 群(2)、ベレー二 アンタル(2,3,4)、南 雅文(1)
1. 北海道大薬医療薬学、2. セゲド大医生理、3. HCEMM-SZTE磁気医療、4. ニューヨーク大神経科学研
*Yuichi Takeuchi(1,2), Qun Li(2), Antal Berenyi(2,3,4), Masabumi Minami(1)
1. Dept Biopharmaceutical Sci Pharmacy, Facult Pharmaceutical Sci, Hokkaido Univ, Sapporo, Japan, 2. Dept Physiol, Facult Med, Univ Szeged, Szeged, Hungary, 3. HCEMM-SZTE Magnetotherapeutics Research Group, Univ Szeged, Szeged, Hungary, 4. Neurosci Inst, New York Univ, New York, USA
Keyword: Decoding, Seizure Susceptibility, Oscillation, Machine Learning
Epilepsy is a neurological disorder characterized by repeated unprovoked seizure occurrences. Fortunately, seizures of up to 70% of epilepsy patients can be successfully controlled by pharmaceutical treatments. However, once seizures are controlled by the anti-epileptic drugs, patients typically need to take the drugs for the rest of their life time; which imposes restrictions on patients in their lives and financial burdens in the society. This issue happens as physicians can not confidently decide when patients shall stop taking the anti-epileptic drugs because there have been no way to properly gage seizure susceptibility of the network in the brain. We here report a new technology to extract and titrate seizure susceptibility from the multi-regional oscillatory brain activity patterns with a semi-supervised machine-learning method called discriminative cross-spectral factor analysis. Adult Long-Evans rats were chronically implanted with 30 ch tungsten wire electrodes in the bilateral hippocampi, the right somatomotor cortices, and the medial entorhinal cortex. The hippocampal commissure was bi-phasically stimulated in six sessions per day for ten days as kindling, a model of temporal lobe epilepsy. The motor seizures typically developed from the third to the fifth day of the kindling period. Local field potentials (LFPs) of the 30 sites during interictal states were recorded in the homecage for one hour everyday before, during, and after the kindling period. Ten cross-spectral factors, three of which were discriminative while the others were residuals, were trained to discriminate the baseline and kindled recordings. Using the trained model (cross-spectral factors), the amount of seizure susceptibility was decoded and quantified with coefficients of the three discriminative cross-spectral factors of linearly projected homecage recordings. The coefficient of the first factor successfully discriminated the baseline and kindled recordings, and titrated the seizure susceptibility. The seizure susceptibility increased in the network from the very first day of the kindling period before the motor seizures developed. These results suggest a neural basis of seizure susceptibility and offer a promising decoding and quantification method of seizure susceptibility. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-110
West症候群の病態における末梢血単球から産生されるIL-1βの関与について
Involvement of peripheral monocytes with IL-1β in the pathogenesis of West syndrome
*高松 朋子(1)、山中 岳(1)、大野 幸子(1)、林 佳奈子(1)、渡邊 由祐(1)、竹下 美佳(1)、鈴木 慎二(1)、森地 振一郎(1)、石田 悠(1)、呉 宗憲(1)、小穴 信吾(1)、柏木 保代(1)、河島 尚志(1)
1. 東京医科大学
*Tomoko Takamatsu(1), Gaku Yamanaka(1), Koko Ohno(1), Kanako Hayashi(1), Yusuke Watanabe(1), Mika Takeshita(1), Shinji Suzuki(1), Shinichiro Morichi(1), Yu Ishida(1), Soken Go(1), Shingo Oana(1), Yasuyo Kashiwagi(1), Hisashi Kawashima(1)
1. Tokyo Medical University
Keyword: infantile spasms, west syndrome, epilepsy, cytokine
West syndrome (WS) is one of developmental and epileptic encephalopathy (DEE) with a poor prognosis, occurring in infancy and often resulting in developmental delay. Although treatments such as ACTH have been shown to be effective, the efficacy of treatment is limited and the pathogenesis of WS is not clarified. Neuroinflammation has been implicated in the pathogenesis of West syndrome (WS). Inflammatory cytokines, including interleukin-1β(IL-1β), have been reported to be associated with epilepsy. However, assessment of cytokine changes in humans is not always simple or deterministic. This study aimed to elucidate the immunological mechanism of WS.
We examined the intracellular cytokine profiles of peripheral blood cells collected from 13 patients with WS using flow cytometry and measured their serum cytokine levels. These were compared with those of 10 age-matched controls. Peripheral blood mononuclear cells were stimulated and cultured with PMA and Ionmycin (lymphocytes), LPS (monocytes), and intracellular cytokines (IL-1β, interleukin-1 receptor antagonist (IL-1RA), interferon gamma (IFN-γ), IL-10, IL-17, Granzyme A, etc.) of CD4+/8+ T cells, B cells/NK cells/NKT cells and monocytes were measured by BD FACS Cant II. In addition, serum cytokine levels were measured using the Bio-plex method and compared with the control group.
We found that the WS group had significantly higher percentages of IL-1β, IL-1RA-positive monocytes, and IFN-γ in CD8+ T cells than the control group. Interestingly, the group with sequelae revealed significantly lower levels of intracellular IFN-γ and IL-6 in CD8+ T and CD4+ T cells, respectively, than did the group without sequelae. There was no correlation between the ratios of positive cells and the serum levels of a particular cytokine in WS patients.
Peripheral immune cells, such as monocytes, that produce IL-1β might be responsible for the pathogenesis of WS. Immunological methods using flow cytometry could facilitate the measurement of IL-1β levels, which remained challenging to assess, thereby contributing to immunological studies of epilepsy. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-111
Spatiotemporal changes in excitatory and inhibitory neuronal activity during seizure progression in a pentylenetetrazol mouse model
*Stern Matthew(1,2)、Berglund Ken(2)、Diamond Jake(2)、Gross Robert(2)
*Matthew A Stern(1,2), Ken Berglund(2), Jake L Diamond(2), Robert E Gross(2)
1. Medical Scientist Training Program, Emory University School of Medicine, Atlanta, GA, USA, 2. Department of Neurosurgery, Emory University School of Medicine, Atlanta, GA, USA
Keyword: Epilepsy, Seizure, Two-Photon Calcium Imaging
Millions of patients worldwide remain without seizure freedom despite an arsenal of treatments at our disposal. A greater understanding of the network alterations underlying seizure activity could inform more targeted approaches to close this treatment gap. Functional optical imaging provides powerful advantages for studying these networks, by enabling the spatiotemporal acquisition of individual neuron activity patterns across multiple seizures induced in mice.Using multiple expression paradigms of genetically encoded calcium indicators to target excitatory and inhibitory cortical cell populations, we performed in vivo two-photon calcium imaging in layer II/III neurons through chronically implanted cranial windows in awake mice, combined with local field potential recording. We recorded multiple imaging sessions within subjects during seizures repeatedly induced by administration of pentylenetetrazol. Examining the pre-ictal to ictal transition, we observe simultaneous excitatory and inhibitory cell recruitment synchronous with pre-ictal spiking. Progressing toward generalized seizure invasion into the imaging field of view, we observe a shift in the relative populations’ activity, with greater and more sustained GABAergic activity ahead of seizure invasion. This could suggest both elevated activity and desynchronization amongst the GABAergic population, reminiscent of inhibitory restraint breakdown. In most seizures imaged, during seizure termination, we observed a slow propagating calcium wave, followed by a sustained decline in calcium signal, consistent with cortical spreading depression. This wave demonstrated a sequential recruitment of neurons and neuropil. Both excitatory and inhibitory cells were recruited simultaneously, reflecting a spread most consistent with a non-synaptic mechanism, such as an extracellular diffusion, gap junction transmission or volume conduction. Furthermore, our analysis demonstrated cortical neuron recruitment during these events within subject followed a similar spatiotemporal pattern across separate seizure episodes. Collectively, these findings are consistent with dynamics observed in several other seizure models, suggesting that cortical seizures may follow a conserved pattern of activity independent of their mode of initiation. Through our continued investigation we hope to gain mechanistic insight that can be translationally leveraged to inform development of novel genetic-based neuromodulatory treatments. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-112
カイニン酸誘導モデルラット潜伏期のSelf-grooming変容に対するオキシトシン受容体の関与
The involvement of oxytocin receptor activation in the change in self-grooming at the latent period in rat kainate model of epilepsy
*新井 博文(1)、夏目 季代久(1)
1. 九州工業大学大学院生命体工学研究科
*Hirofumi Arai(1), Kiyohisa Natsume(1)
1. Kyushu Institute of Technology, Fukuoka, Japan
Keyword: Self-grooming, Correct Transition, Latent period, Epilepsy
The systemic administration of kainate (KA) glutamate receptor agonist induces the epilepsy in rats. The convulsion is induced a few weeks after the administration. It is reported that stereotypy is observed when the rats have convulsions. After the administration day there is a latent period. Whether the stereotypy or the behavioral change is observed at the latent period or not remains unknown. In the previous study, we have reported the self-grooming was facilitated at the period. Self-grooming is an innate behaviour that is involved in hygiene maintenance. The grooming episode includes five stages. Correct transitions (CT) are defined as the transitions between the two stages. Rats have the other transitions than CT. In this study, we studied whether the transitions were modified or not at the latent period. The grooming is related to anxiety feeling. A social bonding-related peptide hormone Oxytocin (Oxt) involves in the feeling. Thirty-six male Wistar rats aged 4-5 weeks were used in the experiment. After anesthesia of 3.5％ isoflurane the rats were divided into KA and Control groups (n = 13 each). KA-group rats were given hourly injections of 0.05 % KA (10 ml/kg, i.p.) until three times. The injected rats had convulsion with more than class III according to a modified Racine’s score. At the latent period from eight to ten days after the KA injection, we recorded using video camera. The number, duration, and CT of the grooming episode were analyzed. The grooming consists of five stages including paw licking (1), nose/face/head grooming (2), body grooming (3), leg grooming (4), and tail and genital grooming (5). CT has the sequential actions from 1 to 5. To study the effect of Oxt receptor, we applied Oxt receptor antagonist atosiban (0.001 %, 10 ml/kg, i.p.) to five KA-group rats for consecutive seven days after the injection. In results, 1) The count, duration, and the ratio of CT in KA group significantly changed at the latent period (Wilcoxon signed rank test (Wsrt); p<0.01 for all parameters), while Control group was not changed. 2) The transition probabilities of 3→4, and 4→5 in CT significantly changed (Wsrt; *p<0.1 and **p<0.01, respectively). 3) When atosiban was applied to the rats, the transitions of 3→4, and 4→5 in CT was suppressed (Wsrt; *p<0.1 for each transition probability). These results suggest that anxiety is decreased with the activation of Oxt receptor at the latent period, and the self-grooming will be facilitated. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-113
Role of hippocampal focal rhythmic discharges (FRDs) in impaired spatial working memory in epileptic mice.
*Pasdarnavab Maryam(1)、Kück Laura(1)、Gerin Fethullah (1)、Ewell Laura(2)
*Maryam Pasdarnavab(1), Laura Kück(1), Fethullah Gerin(1), Laura Ewell(2)
1. University of Bonn, Bonn, Germany , 2. University of California - Irvine, Irvine, USA
Keyword: epilepsy, working memory, hippocampus , memory deficit in epilepsy
Memory impairment is a common cognitive deficit in temporal lobe epilepsy (TLE). The hippocampus is severely altered in TLE exhibiting multiple anatomical changes that lead to a hyperexcitable network capable of generating frequent epileptic discharges and seizures. In this study we investigated whether hippocampal involvement in epileptic activity drives working memory deficits. We employed the supra-hippocampal kainic acid (KA) mouse model of TLE. Consistent with previous reports, histological analysis demonstrated a decrease in CA1 cell layer thickness and granule cell dispersion selectively in the hippocampus ipsilateral to the KA injection (n = 6). Furthermore, mice experienced frequent focal seizures and less frequent behavioural seizures (stage Ⅳ, Racine scale). We observed a working memory deficit in epileptic mice in a delayed-alternation spatial working memory task (Ctrl, 73.54‰ correct; n = 11; KA ,62.07 ‰ correct, n =13). We further investigated the neural mechanisms of the working memory deficit using bilateral LFP recordings from CA1 during task performance and during long sleep sessions pre and post memory task. We discovered that epileptic mice experienced focal rhythmic discharges (FRDs) while they performed the spatial working memory task (n=1686 on the maze, n=7 mice). Spatial correlation analysis revealed that FRDs were often spatially stable on the maze (7/8 mice and 55/124 sessions) – and were most common around reward zones (25 ‰) and delay zones (50 ‰). Preliminary results showed that FRDs shifted to the new reward zones following the change of the reward locations – suggesting that reward-related dynamics such as sharp wave ripple, may drive FRDs. Memory performance was correlated with stability of FRDs (p =< .001, R=0.56), suggesting that spatially unstable FRDs interfere with working memory codes in real time. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-114
新規Kir活性化剤NCC-1566の痙攣モデルにおける抗てんかん作用
Novel inwardly rectifying potassium channel (Kir) activator NCC-1566 on the anti-epileptic effect in the seizure models.
*嶋澤 嘉伸(1)、南谷 武志(1)、野沢 治(1)、伊達 明利(1)、池永 大(2)、野村 岳広(1)、髙橋 大樹(1)、加門 淳司(1)
1. 日産化学株式会社 生物科学研究所 医薬医療材研究部、2. 日産化学株式会社 物質科学研究所 医薬研究部
*Yoshinobu Shimazawa(1), Takeshi Nanya(1), Osamu Nozawa(1), Akitoshi Date(1), Hiroshi Ikenaga(2), Takehiro Nomura(1), Daiki Takahashi(1), Junji Kamon(1)
1. Med Res Dept, Biol Res Labs, Nissan Chemical Corporation, 2. Pharm Res Dept, Chem Res Labs, Nissan Chemical Corporation
Keyword: epilepsy, inwardly rectifying potassium channel , SUDEP
Epilepsy is a common neurological disorder and affects around 50 million people globally. Despite the availability of many antiepileptic drugs, one-third of patients with epilepsy fail to achieve seizure control. In order to create a new anti-epileptic drug that has a new molecular mechanism and a wider safety margin than that of existing medicine, NCC-1566 was discovered by ion-channel assay and in vivo screening. In the present study, we focused on whether glial Kir activator NCC-1566 could relieve seizure behavior.
To begin with, we assessed the audiogenic seizure (AGS) model, maximal electroshock seizure (MES) model, and 6 Hz seizure model. As a result, these seizures were inhibited by NCC-1566. In particular, NCC-1566 suppressed clonic and tonic seizures with ED50 = 5 mg/kg in AGS.
In clinical practice, anti-seizure drugs are often used in combination with control seizures in patients with epilepsy. Therefore, we studied that the anti-convulsant, efficacy of NCC-1566 was evaluated in combination with the anti-seizure drug levetiracetam (LEV) in the 6 Hz model. The 6 Hz models in mice involving 44 mA currents are considered as models of drug resistant epilepsy. Doses of NCC-1566, administered in combination with LEV, were able to shift the 6 Hz 44 mA model ED50 for NCC-1566 by >10-fold. Sudden unexpected death in epilepsy (SUDEP) is the leading cause of death in refractory epilepsy patients and genetic epilepsy patients. Human and animal models suggest that seizure-related respiratory arrest may instigate the cascade that produces cardiorespiratory arrest and death. NCC-1556 was also effective in seizure-related death in the DBA/1 audiogenic mouse model of SUDEP. NCC-1566, in addition to its effect in anti-seizure, did not impair an impairment of rotarod performance with the high dose.
We discovered the first in class novel small molecule glial Kir activator (NCC-1566). Our data suggest that NCC-1566 has an excellent in vivo anti-convulsant, anti-epileptic activity, and safety. NCC-1566 represents the first evidence that the glial Kir activator improves animal models of seizure. NCC-1566 would provide a new treatment option for refractory epilepsy and genetic epilepsy patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-115
細胞接着分子DSCAML1欠損によるてんかん発症の分子機序の解明
Molecular mechanisms of epileptogenesis caused by deficiency of the cell adhesion molecule DSCAML1
*橋詰 晃一(1)、陶山 京香(1)、田谷 真一郎(1)、星野 幹雄(1)
1. 国立精神・神経医療研究センター
*Koichi Hashizume(1), Kyoka Suyama(1), Shinichiro Taya(1), Mikio Hoshino(1)
1. National Center of Neurology and Psychiatry, Tokyo, Japan
Keyword: EPILEPSY, CELL ADHESION
Recent advances in human genetics have led to the identification of a number of genes associated with epilepsy. Although many scientists are analyzing these genes, the mechanism of epileptogenesis remains to be elucidated. Furthermore, many patients are resistant to antiepileptic drugs, which increases the need to develop new therapies for epilepsy. We recently reported that Dscaml1 (Down Syndrome Cell Adhesion Molecule Like 1) is the causative gene of an epileptic rat model, Ihara epilepsy rat. DSCAML1 is a cell adhesion molecule with 10 immunoglobulin (Ig) domains and 6 fibronectin type III domains, and its structure is similar to that of the closely related DSCAM. Although there is a growing body of knowledge that DSCAM/DSCAML1 regulates neuronal tiling and neurite outgrowth, its relevance to epilepsy remains largely unknown. To elucidate the molecular mechanisms underlying epileptogenesis caused by the loss of DSCAML1, we performed gain- and loss-of-function experiments. We found that Dscaml1 knockdown in primary hippocampal neurons increased the level of phosphorylated ribosomal protein S6 (RPS6), which is associated with epileptogenesis. Consistent with this result, exogenous expression of Dscaml1 in fibroblasts decreased the phosphorylation level of RPS6. Furthermore, it was suggested that DSCAML1 regulates the phosphorylation of RPS6 in a manner dependent on the nuclear translocation of the intracellular domain. Since the intracellular domain of DSCAML1, together with signaling molecules, has been reported to regulate transcription, these results suggest that DSCAML1 negatively regulates RPS6 and epilepsy through transcriptional control. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-116
TSC欠損型ヒト神経細胞はmTOR依存的にL-typeカルシウムチャネルのカルシウム流入増加を引き起こす
TSC inactivation increases neuronal network activity by enhancing Ca(2+) influx via L-type Ca(2+) channels
*久恒 智博(1)、島田 忠之(1)、宮本 章歳(2)、Amy Lee(3)、山形 要人(1)
1. 東京都医学総合研究所、2. 京都大学 生命科学研究科 分子動態生理学、3. Dept. of Neuroscience, University of Texas-Austin
*Chihiro Hisatsune Hisatsune(1), Tadayuki Shimada(1), Akitoshi Miyamoto(2), Amy Lee(3), kanato Yamagata(1)
1. The Tokyo Metropolitan Institute of Medical Science , 2. Laboratory of Single-Molecule Cell Biology, Kyoto University Graduate School of Biostudies, 3. Dept. of Neuroscience, University of Texas-Austin
Keyword: TSC, epilepsy, Ca2+ signal
Tuberous sclerosis complex (TSC) is a multisystem developmental disorder characterized by hamartomas in various organs including the brain, lungs, and kidneys. The patients with TSC present neurological impairments such as epilepsy, intellectual disability, and autism. Among them, epilepsy is a major concern and has an intimate relationship with developmental outcomes and quality of life. Sustained activation of mammalian target of rapamycin (mTOR) by either TSC1 or TSC2 mutation is thought to be involved in epilepsy onset in TSC. However, the mechanism by which mTOR causes seizures remain unknown.
To examine the possible mechanism of epilepsy onset in TSC, we developed neurons from human iPS cells with TSC2 mutation and analyzed their intracellular Ca2+ dynamics by Ca2+ imaging. We found that cultured TSC2-deficient neurons exhibited highly synchronized neuronal activity. Moreover, TSC2-deficient neurons presented enhanced Ca2+ influx via L-type Ca2+ channels (LTCCs), which contributed to the abnormal neurite extension and sustained activation of cAMP response element binding protein (CREB), a critical mediator of synaptic plasticity. Expression of Cav1.3, a subtype of LTCCs, was increased in TSC2-deficient neurons, but long-term rapamycin treatment suppressed this increase and reversed the altered neuronal activity and neurite extensions. Thus, we identified Cav1.3 LTCC as a critical downstream component of TSC-mTOR signaling that would trigger enhanced neuronal network activity of TSC2-deficient neurons. We suggest that LTCCs could be potential novel targets for the treatment of epilepsy in TSC. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-117
ペリサイトマーカーによるWest症候群の免疫学的検討
Involvement of pericytes in the pathogenesis of West syndrome
*渡邊 由祐(1)、山中 岳(1)、林 佳奈子(1)、鈴木 慎二(1)、竹下 美佳(1)、森下 那月美(1)、森地 振一郎(1)、呉 宗憲(1)、石田 悠(1)、小穴 信吾(1)、高田 芙友子(2)、柏木 保代(1)、河島 尚志(1)
1. 東京医科大学小児科思春期科学分野、2. 福岡大学薬学部薬学疾患管理学教室
*Yusuke Watanabe(1), Gaku Yamanaka(1), Kanako Hayashi(1), Shinji Suzuki(1), Mika Takeshita(1), Natsumi Morishita(1), Shinichiro Morichi(1), Soupei Go(1), Yu Ishida(1), Shingo Oana(1), Fuyuko Takata(2), Yasuyo Kashiwagi(1), Hisashi Kawashima(1)
1. Department of Pediatrics and Adolescent Medicine, Tokyo Medical University, Tokyo, Japan, 2. Department of Pharmaceutical Care and Health Sciences, Faculty of Pharmaceutical Sciences, Fukuoka University, Fukuoka, Japan
Keyword: West syndrome, pericyte, PDGFRβ , CD13
Background: There is significant evidence that the pathogenesis of epilepsy is related to neuroinflammation and cerebrovascular dysfunction. In addition, previous studies have reported the relationship between neuroinflammation and West syndrome (WS). Pericytes play both roles, namely maintenance of the blood-brain barrier and involvement in neuroinflammation, attracting attention as to whether they are also involved in the pathogenesis of epilepsy. We believe that evaluating the relationship between WS, whose etiology remains unknown, and pericytes, which play an important role in the central nervous system, will lead to new therapeutic targets for WS. Aim: To explore the relationship between WS and pericytes. Methods: Eighteen Japanese pediatric patients with WS and nine controls aged 2 years or younger were retrospectively enrolled in this study. We assessed the serum levels of pericyte markers, plasma PDGFRβ (platelet-derived growth factor receptor β), CD13 (aminopeptidase N) by using enzyme-linked immunosorbent assay, and 27 cytokines by using multiplex cytokine assay in 18 pediatric patients with WS and control groups. Results: Patients with WS exhibited significantly increased level of CD13 and decreased level of PDGFRβ compared to controls, but no difference in serum cytokine levels. These values showed no significant difference between symptomatic and idiopathic WS. Discussion: High levels of CD13 expression in pericytes may cause angiogenesis and subsequently contribute to the pathogenesis of WS. Decreased PDGFRβ levels in WS may lead to dysregulation of pericyte survival, proliferation, and migration signals, and these results may be secondary to endothelial dysfunction caused by CD13 elevation. On the other hand, we did not find any association between 27 cytokines and pericyte markers. Conclusion: Pericytes might be implicated in the pathogenesis of WS. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-118
GATOR1関連てんかんモデル動物におけるラパマイシンの発作抑制効果の検討
Different rapamycin susceptibility among epileptic animal models of GATORopathy
*趙 迪(1,2)、澤田 裕太(1)、平岡 優一(1)、真下 知士(2)、田中 光一(1)、石田 紗恵子(1,2)
1. 東京医科歯科大学、2. 東京大学
*DI ZHAO(1,2), Yuta SAWADA(1), Yuichi HIRAOKA(1), Tomoji MASHIMO(2), Kohichi TANAKA(1), Saeko ISHIDA(1,2)
1. Tokyo Medical and Dental University, 2. The University of Tokyo
Keyword: rapamycin, GATOR1, animal model, mTOR
Variations in the GATOR1 (GAP activity towards RAGs 1) complex genes (including DEPDC5, NPRL2, and NPRL3) are involved in familial focal epilepsies. Since the variations are frequent and have been implicated to associate with a broad spectrum of focal epilepsies, a unique pathology “GATORopathy” is categorized. GATOR1 is a repressor of the mTORC1 (mechanistic target of rapamycin complex 1) pathway, which regulates major functions such as protein synthesis and cell growth. Since upregulation of mTORC1 activity has been observed in the patients, we examined whether administration of the mTOR inhibitor, rapamycin, prevent spontaneous seizures of animal models of GATORopathy. Rapamycin or vehicle was chronically injected to dorsal telencephalon-specific Depdc5-, Nprl2- and Nprl3-knockout (Depdc5-cKO, Nprl2-cKO, Nprl3-cKO) mice from P13 to P42. mTORC1 activity was down regulated by the rapamycin treatment in Nprl2-, Nprl3-, and Depdc5-cKO mice. Continuous video recording revealed a significant reduction in both the number and duration of generalized seizures in rapamycin-treated cKO mice compared with vehicle-treated cKO mice. Lifespan of each rapamycin-treated cKO mouse line was significantly prolonged compared with each vehicle-treated cKO mouse line. 71% of Depdc5-cKO mice survived longer than P80 after withdrawal at P42. However, all the Nprl2- and Nprl3-cKO mice died significantly earlier than Depdc5-cKO mice. Video recording revealed that all Nprl2- and Nprl3-cKO mice died either during generalized seizures or in the recovery period after withdrawal at P42. It suggests that the benefit of rapamycin after withdrawal was less durable in Nprl2- and Nprl3-cKO mice compared with Depdc5-cKO mice. Further studies using these conditional knockout mice will be useful for understanding GATORopathy and for the identification of novel therapeutic targets. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-119
Glial cells missing 1 は脳損傷コア領域のグリア細胞分化と血管新生を誘導する
Glial cells missing 1 induce glial cells and angiogenesis in the core region of brain injury
*林 義剛(1)、郷 康広(2)、中林 一彦(3)、アブドゥラ アスマ(1)、モハマド ザキヤ(1)、等 誠司(1)
1. 滋賀医科大学 統合臓器生理学、2. 自然科学研究機構 生命創成探究センター、3. 国立成育医療研究センター研究所 周産期病態研究部
*Yoshitaka Hayashi(1), Yasuhiro Go(2), Kazuhiko Nakabayashi(3), Asmaa Abdullah(1), Zakiyyah Mohd(1), Seiji Hitoshi(1)
1. Integrative physiology, Shiga University of Medical Science, Shiga, Japan, 2. Exploratory Research Center on Life and Living Systems (ExCELLS), National Institutes of Natural Sciences, Aichi, Japan, 3. Department of Maternal-Fetal Biology, National Center for Health and Development, Tokyo, Japan
Keyword: Brain injury, astorocyte, oligodendrocyte, angiogenesis
Glial cell missing (gcm) plays a critical role in glial cell development in Drosophila. However, the function of Gcm1 in the mammalian brain remains to be investigated because Gcm1-deficient mice are embryonic lethal around embryonic day 10. Using an in utero electroporation, we revealed that overexpression of Gcm1 in the brain of mammalian embryos promotes the differentiation of neural progenitor cells into astrocytes and oligodendrocytes. Furthermore, those brains were also shown to promote the angiogenesis around Gcm1 expressing cells. We found that Lif and Vegf are involved in astrocyte differentiation and angiogenesis, respectively, by RNA-seq and knockdown studies. On the other hand, when the brain is injured, the proliferation of glial cells and angiogenesis are promoted and play an important role in repair. To clarify the relationship between Gcm1 and brain damage, we studied a cryo brain injury model using wild type and heterozygous Gcm1 knockout mice (Gcm1 het) and quantified gene expression level in the brain on days 1, 2, 3, 5, 7, and 14 after injury. The results showed that Gcm1 expression was upregulated in the brain 1-2 days after injury and suppressed in the Gcm1 het. Furthermore, the expression of Lif, Vegfc, angiogenesis-related genes (Angpt1), and neural progenitor-related gene (Nestin) was significantly decreased in Gcm1 het compared to wild type at 1-3 days after injury, while the expression of the astrocyte-related gene (Gfap) and oligodendrocyte-related genes (Olig2, Ng2) was not. In the core region of injury, we observed Gcm1-positive cells attached to neovessels and reduced co-localization of astrocytes in neovessels. These results suggest that Gcm1 induces neuro-gliogenesis and angiogenesis in the core region of brain injury through the secretion of growth factors. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-120
外傷性脳損傷の新規モデルを用いたタウオパチー病理研究
Tauopathy pathology in a novel traumatic mouse brain injury model
*木村 妙子(1)、鈴掛 雅美(1)、橋本 雅史(1)、久永 眞市(1,2)、長谷川 成人(1)
1. 公益財団法人東京都医学総合研究所、2. 東京都立大学大学院理学研究科
*Taeko Kimura(1), Masami Suzukake(1), Masashi Hashimoto(1), Shin-ichi Hisanaga(1,2), Masato Hasegawa(1)
1. Tokyo Metropolitan Institute of Medical Science, 2. Department of Biological Sciences, Tokyo Metropolitan University
Keyword: traumatic brain injury, Tauopathy, mouse model, tau
Tauopathy is a group of neurodegenerative diseases, including Alzheimer's disease (AD), characterized by aggregates of tau in neurons and glial cells. The pathological mechanism of tauopathy is not known yet. Recently, chronic traumatic brain injury (CTE) caused by traumatic brain injury (TBI) is shown to be also a tauopathy. Different from other tauopathies, we can follow the disease progression of CTE because the onset and cause are clear. However, most previous CTE models are invasive and did not represent sports injuries and traffic accident injuries. Here, we used a novel TBI model called CHIMERA (closed-head impact model of engineered rotational acceleration). Firstly, we established TBI conditions that could cause tau pathology using wild-type mice. With mild repeated TBI, mice displayed axonal damages such as varicosities and bulbs in optic tract at 7 days after TBI. Iba1 and GFAP-positive inflammation was also observed at the axon injury sites, but hyperphosphorylated tau (AT8) positive pathology was not. Then, we used tau Tg (Tau Tg) mice, which express tau with pathological Ser mutation at Pro301 and accumulate tangles of hyperphosphorylated tau at 6 months. Tau Tg mice also showed axonal inflammation at 1~2 months after TBI, whereas tau pathology was not observed. In contrast, Tau Tg mice increased tau hyperphosphorylation at the IV and V layers in the motor and sensory cortex, where human TBI has tau pathology. The IV, V layer neurons extends long axons to spinal cord. We are now studying whether their long axons are damaged by TBI. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-121
ラットの予防運動は、酸化ストレス、炎症、神経細胞のアポトーシスを抑制し、くも膜下出血による早期脳損傷を減少させる。
Preconditioning exercise in rats attenuates early brain injury resulting from subarachnoid hemorrhage by reducing oxidative stress, inflammation, and neuronal apoptosis.
*大塚 章太郎(1)、松岡 輝樹(2)、松崎 凌真(2)、中古川 智美(2)、谷 明(2)、則松 貢輔(2)、 高田 聖也(2)、中西 和毅(2)、丸山 征郎 (1)、榊間 春利 (2)、菊池 清志(1,3)
1. 鹿児島大学大学院医歯学総合研究科、2. 鹿児島大学大学院保健学研究科、3. 久留米大学医学部医学科 生理学講座 脳・神経機能部門
*Shotaro Otsuka(1), Teruki Matsuoka(2), Ryoma Matsuzaki(2), Tomomi Nakakogawa(2), Akira Tani(2), Kosuke Norimatsu(2), Seiya Takada(2), Kazuki Nakanishi(2), Ikuro Maruyama (1), Harutoshi Sakakima(2), Kiyoshi Kikuchi(1,3)
1. Kagoshima University Graduate School Medical and Dental Science , 2. Department of Physical Therapy, School of Health Science, Faculty of Medicine, Kagoshima University, 3. Division of Brain Science, Department of Physiology, Kurume University School of Medicine
Keyword: Preconditioning exercise, Subarachnoid hemorrhage, Early brain injury, Neuroprotective effects
Subarachnoid hemorrhage (SAH) is a catastrophic form of stroke responsible for significant morbidity and mortality. Oxidative stress, inflammation, and neuronal apoptosis are important in the pathogenesis of early brain injury (EBI) following SAH. Preconditioning exercise confers neuroprotective effects, mitigating EBI; however, the basis for such protection is unknown. We investigated the effects of preconditioning exercise on brain damage and sensorimotor function after SAH. Forty-eight male Sprague Dawley (SD) rats, 5 weeks old, with mean body weight 240–260 g, were used in this study. SD rats were assigned to either a sham-operated (Sham, n = 8) group, exercise (Ex, n = 20) group, or no-exercise (No-Ex, n = 20) group. After a 3-week exercise program, they underwent SAH by endovascular perforation. Consciousness level, neurological score, and sensorimotor function were studied. The expression of nuclear factor erythroid 2 p45-related factor 2 (Nrf2), heme oxygenase 1 (HO-1), 4-hydroxynonenal (4HNE), nitrotyrosine (NT), ionized calcium-binding adaptor molecule 1 (Iba1), tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6), interleukin 1β (IL-1β), 14-3-3γ, p-β-catenin Ser37, Bax, and caspase-3 were evaluated by immunohistochemistry or western blotting. The terminal deoxynucleotidyl transferase-mediated biotinylated dUTP nick end labeling (TUNEL) assay was also performed. After SAH, the Ex group had significantly reduced neurological deficits, sensorimotor dysfunction, and consciousness disorder compared with the No-Ex group. Nrf2, HO-1, and 14-3-3γ were significantly higher in the Ex group, while 4HNE, NT, Iba1, TNF-α, IL-6, IL-1β, Bax, caspase-3, and TUNEL-positive cells were significantly lower. Following a 3-week regimen of preconditioning exercise, we observed that neurological scores as well as sensorimotor dysfunction and consciousness disorders were improved after SAH by reduced EBI. As a mechanism, 4HNE and NT expression was reduced by activating the Nrf2/HO-1 pathway, and oxidative stress was suppressed. Additionally, the inhibition of proinflammatory cytokines reduced inflammation and apoptosis, likely via the 14–3-3γ/p-β-catenin Ser37/Bax/caspase-3 pathway. On the basis of these results, we suggest that preconditioning exercise is an effective prophylactic method for attenuating the effects of SAH and can reduce the severity of EBI. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-122
脳損傷は異なる時間経過で細胞タイプに特徴的な小胞体ストレス応答を惹起する
Brain injury triggers cell-type-specific endoplasmic reticulum stress responses in different time courses.
*宝田 美佳(1)、範 啓琰(1)、沖谷 なほ子(1)、堀 修(1)
1. 金沢大学医薬保健研究域医学系
*Mika Takarada-Iemata(1), Qiyan Fan(1), Nahoko Okitani(1), Osamu Hori(1)
1. Grad Sch Med Sci, Kanazawa Univ, Ishikawa, Japan
Keyword: ER stress, brain injury, astrocytes, endothelial cells
Endoplasmic reticulum (ER) stress is caused by the accumulation of misfolded/unfolded proteins in the ER lumen. As a response to improve the situation of the accumulation of misfolded proteins in the ER, cells induce a series of signal transcription pathway termed the unfolded protein response (UPR). The UPR coordinates protein homeostasis by regulating protein synthesis, quality control and degradation, but also triggers cell death when these adaptive responses fail to improve protein homeostasis. Accumulating evidence suggests that the UPR plays an important role in neuropathological conditions such as neurodegenerative diseases and brain insults. However, how ER stress is actually induced under pathological conditions in vivo, including the cell types and timing of the UPR induction, is not fully understood. In this study, we investigated the cell types and temporal patterns of the ER stress response after brain injury using the cortical stab injury model in ER stress-activated indicator (ERAI) mice. ERAI mice enable visualization of ER stress by detecting the activation of X-box binding protein 1, a transducer of the UPR, as a fluorescence of fusion protein with Venus. The fluorescent signals and the number of signal-positive cells increased over time in the ipsilateral cortex with time after brain injury in ERAI mice. The reporter signals were observed in injured neurons in the early times after brain injury. However, the majority of the cells positive for the fluorescent signals were non-neuronal cells such as vascular cells and astrocytes throughout the period analyzed after brain injury. These results suggest that UPR may play important roles not only in neurons but also in the non-neuronal cells in the course of neurological diseases including brain injury. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-123
自発的運動は高齢期に失われる脳損傷後の皮質脊髄路の再編と運動機能を回復させる
Voluntary exercise restores the ability of corticospinal tract rewiring and motor recovery loss in aged mice after brain injury
*田中 貴士(1,2)、浦 大樹(3)、硎 澄仁(3)、新井田 要(3)、上野 将紀(4)
1. 熊本保健科学大学保健科学部、2. 金沢医科大学医学部、3. 金沢医科大学総合医学研究所、4. 新潟大学脳研究所
*Takashi TANAKA(1,2), Hiroki URA(3), Sumihito TOGI(3), Yo NIIDA(3), Masaki UENO(4)
1. Grad Sch Health Sci, Kumamoto Health Sci Univ, Kumamoto, Japan, 2. Sch Med, Kanazawa Med Univ, Ishikawa, Japan, 3. Med Res Inst, Kanazawa Med Univ, Ishikawa, Japan, 4. Brain Res Inst, Niigata Univ, Niigata, Japan
Keyword: Brain injury, Aged mice, Voluntary exercise, Motor recovery
Brain injury often causes severe motor dysfunction, which makes it difficult to maintain an independent social life. Various therapeutic interventions, such as molecular targeting and rehabilitative motor exercise, have been investigated in promoting the reorganization of remnant neural networks and functional recovery after brain injury. We previously showed that the axons of the corticospinal tract (CST) are rewired to create an intraspinal circuit that contributes to motor recovery after brain injury in young adult mice. However, few studies have evaluated the ability of rewiring and functional recovery in aged rodents. In this study, we examined the rewiring efficiency of the corticospinal axons for functional recovery in aged mice and further investigated whether rehabilitative motor training could promote functional recovery after brain injury. For this purpose, sprouting of the CST axons originating from the contralesional motor cortex was analyzed using neuronal tracers following unilateral cortical injury in young adult and aged mice with⁄without voluntary running exercise. We found that sprouting of the CST axons after cortical injury was mostly not induced in the aged mice, which was in contrast to that observed in the young adult mice. However, voluntary exercise significantly increased axon sprouting and enhanced motor recovery in the aged mice, which were comparable to those levels observed in the young adult mice. Next, we performed RNA sequencing to compare the transcriptomes of the contralesional motor cortices of the aged mice with those of the young adult mice. We found that 271 genes that had increased expression in the aged group showed decreased expression after exercise, whereas 20 genes that exhibited decreased expression in the aged group demonstrated increased expression after exercise. These gene expression changes indicated that exercise in the aged group contributed to the return to young gene expression levels. In conclusion, our study revealed that voluntary exercise restored the ability of CST sprouting and motor recovery in aged mice, similar to the levels induced in young adult mice, along with robust gene expression changes in the motor cortex. These findings suggest that voluntary exercise could rejuvenate the aged brain to promote neural plasticity after brain injury. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-124
温熱及び超音波照射による新規脳血栓溶解療法の有効性と安全性の検討
Efficacy and safety of novel thrombolysis with spot heating and ultrasound irradiation
*森原 隆太(1)、山下 徹(1)、福井 裕介(1)、小坂田 陽介(1)、田所 功(1)、武本 麻美(1)、馮 田(1)、胡 欣冉(1)、阿部 康二(1)
1. 岡山大学病院
*Ryuta Morihara(1), Toru Yamashita(1), Yusuke Fukui(1), Yosuke Osakada(1), Koh Tadokoro(1), Mami Takemoto(1), Tian Feng(1), Xinran Hu(1), Koji Abe(1)
1. Okayama University
Keyword: sonothrombolysis, heating, sonication, endothelium dysfunction
Background and Purpose— The feasibility of transcranial sonothrombolysis has been demonstrated, although little is known about the relationships between thermal or mechanical mechanisms and thrombolytic outcomes. The present study aims to reveal the effect and safety of temperature and ultrasound through in vitro and in vivo thrombolysis models. Methods— Artificial clots were prepared by using fibrin glue, Bolheal, which is composed of fibrinogen and thrombin. Then these clots in microtubes were heated in a water bath or sonicated by ultrasound irradiation. The change in weight of clots was then measured. In an in vitro model to mimic thrombotic occlusion, clots were exposed to a spot heater until recanalization occurred. In an in vivo study, the left common carotid artery (CCA) of Wistar and SHR rats was exposed to a spot heater with 55 degrees or sonication with 20kHz. In addition to Nissl stain and immunohistochemistry of brain, hematoxylin and eosin staining and immunohistochemistry of CCA were performed at 1 and 7 days. Results— Clot weight decreased proportionally with rising temperature and sonication time. In the in vitro thrombotic occlusion model, based on spot heating, we observed clots volume reduction and clots moving to distal side, followed by recanalization of occlusion at around 55 degrees within 5 min. After heating the CCA at 55 degrees for 2 or 5 min or sonication with 20kHz for 2 or 5 sec, no brain infarct, brain blood barrier disruption, astrocyte responses, or microglial activation was shown, but the loss of endothelial junctional integrity and an inflammatory response in the carotid artery were detected. Conclusions— The present spot heating and ultrasound irradiation models seem to be effective for disintegrating clots in vitro, but the safety of the in vivo model was not fully supported by the data. However, the data indicates that a shorter time exposure could be less invasive than a longer exposure. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-125
若齢及び老齢オスラット局所脳虚血モデルにおけるプロゲステロン受容体アゴニストnestoroneの長期神経保護作用
Long-term neuroprotection of the progesterone receptor agonist nestorone against permanent focal cerebral ischemia in adult and old male rats.
*田中 基樹(1)、曽我部 正博(2)、浅井 真人(1)
1. 愛知県医療療育総合センター発達障害研究所、2. 名古屋大学大学院医学系研究科
*Motoki Tanaka(1), Masahiro Sokabe(2), Masato Asai(1)
1. Inst for Dev Res, Aichi Dev Disabil Ctr, kasugai, Japan , 2. Nagoya Univ Grad Sch Med, Nagoya, Japan
Keyword: neuroactive steroid, neuroprotection, stroke
Ischemic stroke is a leading cause of death and disability in a large number of countries. Thrombolysis by tissue plasminogen activator is the most effective treatment currently available, while it has limitations such as a very narrow therapeutic time-window (4.5 h after symptom onset) and serious adverse effects. Progesterone (P4) has been reported to exert long-term neuroprotection against transient and permanent focal cerebral ischemia in adult and old rodents even by post-ischemic administration long after the incident, and P4 receptors are essential for the neuroprotection. However, P4 exhibits androgenic, estrogenic, and GABAergic activities via several of its metabolites, which may bring about unfavorable effects in human patients. Nestorone is a highly potent selective P4 receptor agonist without such steroidogenic and GABAergic activities. Our recent study demonstrated that sole activation of P4 receptors by nestorone is sufficient to exert long-term neuroprotection against transient focal cerebral ischemia in adult male rats. The present study examined effects of nestorone on permanent focal cerebral ischemia in both adult and old male rats which are better models to evaluate outcomes in treatments for typical stroke patients. Adult (6-month-old) or old (18-month-old) male rats were subjected to permanent focal cerebral ischemia by permanently occluding the left middle cerebral artery (pMCAO) and transiently bilateral common carotid arteries. Nestorone (10 μg/kg/day) or its vehicle (30% hydroxypropyl-β-cyclodextrin) was continuously administered during 7 days by Alzet osmotic pumps, starting at 18 h after pMCAO. Nestorone-treated adult male rats showed marked improvements of functional outcomes in the adhesive removal and Rotarod tests compared with vehicle-treated ones 30 days after pMCAO. Consistent with the functional improvements, nestorone significantly reduced infarct sizes 30 days after pMCAO in adult male rats. The same administration way of nestorone provided comparable functional and histological improvements in old male rats. These results suggest that nestorone exerts robust long-term neuroprotection against permanent focal cerebral ischemia in both adult and old male rats. Nestorone is thus a promising agent in post-stroke treatment due to its age-independent wide therapeutic time-window (18 h after symptom onset) and potent progestational effects without other steroid-related activities. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-126
ラット脳内出血モデルにおける感覚運動障害の回復と脚内核の残存容積は正の相関を示す
Recovery from sensory-motor dysfunctions positively correlates with a residual volume of the entopeduncular nucleus in a rat model of the intracerebral hemorrhage.
*酒井 涼(1,2)、村田 航志(2)、黒田 一樹(2)、領家 崇(2)、深澤 有吾(2)
1. 福井医療大学、2. 福井大学学術研究院医学系研究科脳形態機能学分野
*Ryo Sakai(1,2), Koshi Murata(2), Kazuki Kuroda(2), Takashi Ryoke(2), Yugo Fukazawa(2)
1. Dept Reha, Fukui Health Science Univ, Fukui, Japan, 2. Dept Med, Univ of Fukui, Fukui, Japan
Keyword: intracerebral hemorrhage, senrosy-motor dysfunction, entopeduncular nucleus, reaching movement
Intracerebral hemorrhage (ICH) is one of the most common diseases in clinical practice, and animal models that reproduce the pathology have been used to investigate mechanisms underlying functional recovery from the stroke. However, how the location and size of the injured area affect the functional impairment and its recovery have not been thoroughly investigated. In the present study, we induced ICH in the rat internal capsule (IC) and investigated the relationship between the location and volume of ICH, the severity of the impairment on the sensory-motor functions, and the recovery. ICH was induced by the stereotaxic injection of collagenase type IV, which lyses the basement membrane of blood vessels, into the IC of 11 Long-Evans rats. The injured areas were categorized as the internal capsule, the dorsomedial and ventrolateral regions to the IC, and the entopeduncular nucleus (EP), and their volumes were measured. To quantify the damage on motor function of the forelimb, the single pellet reaching task test was carried out and scored before and 2, 7, 14, 21, and 28 days after the injection. In addition, the error patterns of the reaching movement were classified. The von Frey test was carried out on 2 and 28 days after the injection for the sensory function. While the motor and sensory functions significantly declined on postoperative day 2 in all rats, the functional recovery within 28 days was variable. Most of the hemorrhage was found within the IC in all individuals, but some were extended to the surrounding area. The total volume of damaged tissue, the damaged volumes in the IC, and the dorsal medial and ventral lateral regions to the IC did not correlate with the severity of motor or sensory impairment. On the other hand, motor and sensory function on postoperative day 28 was significantly correlated with the volume of remaining EP. In addition, although the preoperational rats failed to obtain food pellets primarily by "grasp failure” during the reaching task of the forelimb, the rats on postoperative 28 mostly failed by "initial or final trajectory error" and the frequency of these errors was negatively correlated with the volume of remaining EP significantly. No correlation was detected between the volume of hemorrhage and the EP damage. Our study suggests that EP plays a pivotal role in the recovery of sensory-motor functions after cerebral hemorrhage. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-127
慢性高血圧ラットにおいて一過性増殖能を獲得した血管周囲ミクログリアは脳血管障害に先んじて炎症性形質を呈する
Transiently proliferating vessel-associated microglia harbor pro-inflammatory and precede cerebrovascular changes in a chronic hypertension rat
*小泉 崇(1,3)、田口 勝敏(1)、水田 依久子(3)、鳥羽 裕恵(2)、大西 興洋(4)、生駒 和也(4)、中田 徹男(2)、水野 敏樹(3)、田中 雅樹(1)
1. 京都府立医科大学大学院医学系研究科 生体構造科学部門、2. 京都薬科大学病態薬科学系臨床薬理学分野、3. 京都府立医科大学大学院医学系研究科 脳神経内科学、4. 京都府立医科大学大学院医学系研究科 整形外科学
*Takashi Koizumi(1,3), Katsutoshi Taguchi(1), Ikuko Mizuta(3), Hiroe Toba(2), Okihiro Onishi(4), Kazuya Ikoma(4), Tetsuo Nakata(2), Toshiki Mizuno(3), Masaki Tanaka(1)
1. Dept of Anatomy and Neurobiology, Grad Sch Med, Kyoto Prefectural University of Medicine, Kyoto, Japan, 2. Dept of Clinical Pharmacology, Div of Pathological Sciences, Kyoto Pharmaceutical University, Kyoto, Japan., 3. Dept of Neurology, Grad Sch Med, Kyoto Prefectural University of Medicine, Kyoto, Japan, 4. Dept of Orthopaedics, Grad Sch Med, Kyoto Prefectural University of Medicine, Kyoto, Japan
Keyword: hypertension, vessel-associated microglia, cerebral small vessel disease
Background: Microglia play a crucial role in the maintenance of parenchymal brain homeostasis. Activated microglia show a biphasic influence, promoting beneficial repair and causing harmful damage. It is well-known that microglia are initially activated to the anti-inflammatory M2 state and subsequently switch to the pro-inflammatory M1 state, called M2-to-M1 class switching, in acute ischemic models. However, the activation process of microglia in chronic and sporadic hypertension remains poorly understood, though hypertension is one of the essential factors for cerebral small vessel disease. Objects: We aimed to clarify the process of microglia dynamics using a chronic hypertension model, the deoxycorticosterone acetate (DOCA)-salt-treated Wistar rats. Methods: After unilateral nephrectomy, the rats were randomly divided into DOCA-salt and control groups. DOCA-salt rats received a weekly subcutaneous injection of DOCA (40 mg/kg) and were continuously provided with 1% NaCl in drinking water. Control rats received no administration of DOCA and were provided with tap water. To investigate the temporal expression profiles of pro-inflammatory and anti-inflammatory markers for microglia, the animals were subjected to immunohistochemical and biochemical studies after 2, 3, or 4 weeks of DOCA-salt treatment. Results: Hypertension occurred after 2 weeks of DOCA and salt treatment, when activated shaped microglia juxtaposed to the vessels appeared, although the histopathological findings were normal. After 3 weeks of DOCA and salt treatment, proinflammatory-state vessel-associated and parenchymal microglia significantly increased, when local histopathological findings began to be observed but cerebrovascular destruction did not occur. On the other hand, anti-inflammatory state microglia were never observed around the vessels through any periods. Interestingly, only at the 2 and 3 weeks of DOCA and salt treatment, vessel-associated and parenchymal microglia significantly expressed Ki-67, one of the cell proliferation markers. Conclusions: We concluded that the vessel-associated microglia increased followed by switching to the pro-inflammatory state, via a transient proliferative state, before hypertension-induced cerebrovascular disruptions. Our results suggest that proliferative vessel-associated microglia could have the potential to regulate vascular physiology and work as initial key players to the development of hypertension-induced cerebral small vessel disease. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-128
Automatic Diagnosis, Classification and Subtyping of Ischemic Stroke based on a Deep Learning System
*Wei SHAN(1,2,3), Huajun Yang(1), He Fu(2), Zhenzhou Wu(2), Qun Wang(1,2,3)
1. Beijing Tiantan Hospital, Capital Medical University, Beijing, China, 2. National Center for Clinical Medicine of Neurological Diseases, Beijing, 100070, P.R. China, 3. Beijing Institute for Brain Disorders, Beijing, 100070, P.R. China
Keyword: Diagnosis, Classification, Ischemic Stroke, Deep Learning System
Objectives: To validate the efficiency and reliability of an automatic tool based on a well-established deep learning system (DLS) for the diagnosis, classification, and subtyping of ischemic stroke. Methods: Four deep learning models in this study are designed based on a convolutional neural network, trained and validated based on 6,000 patients diagnosed with ischemic stroke, after screening, finally, 2,859 patients’ DWI images (1.5T) and 2,877 patients’ DWI images (3.0T). The blood Supply Model is established according to the arterial territories of the whole brain demonstrating the cerebral blood supply and the Border Zone Model is furtherly focused on the watershed regions, both of which are used for automatic classification and subtyping. On the other hand, the Brain Structure Atlas Model is designed on the anatomical sub-regions of the whole brain, and Cortex Model is established focusing on cortical subdivisions, both of which are used for anatomical diagnosis of ischemic stroke. The models are then tested with 100 patients’ images and the performance is compared with that of 2 neurologists with at least 15 years experience. These results are evaluated by these two neuroradiologists and given an output analysis score, which are divided into 2 grades (accepted=1, not accepted=0). The inter-neuroradiologists agreement rate, assessed by the Kendall-W test, is used for quality analysis. Results: The results show that all the four appropriately trained deep learning models could achieve a high level of accuracy. The Dice coefficient is 0.71 in Blood Supply Model, 0.79 in Border Zone Model, 0.77 in the Brain Structure Atlas Model and 0.81 in the Brain Cortex Model. The performance of the models is comparable to the experienced neurologists, while automatic diagnosis, classification and subtyping by the deep learning models are much helpful to the neurologists. The neuroradiologists report that about 90% of the results could be directly accepted in clinical evaluation tests. Conclusions: The result of this study shows that the automatic diagnosis, classification, and subtyping of ischemic stroke are practicable based on a well-trained deep learning system. The proposed DLS in this study could be trusted as a promising tool for the diagnosis and investigation of ischemic stroke and could be further evaluated in other brain disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-129
ホヤ由来プラズマローゲンは脳梗塞モデルマウスの酸化ストレス・炎症を抑制する
Antioxidative and neuroprotective effects of ascidiacea-derived plasmalogen in a mouse stroke model
*福井 裕介(1)、中野 由美子(1)、柚木 太淳(1)、武本 麻美(1)、森原 隆太(1)、山下 徹(1)、阿部 康二(2)
1. 岡山大学大学院医歯薬学総合研究科、2. 国立精神・神経医療研究センター
*Yusuke Fukui(1), Yumiko Nakano(1), Taijun Yunoki(1), Takemoto Mami(1), Ryuta Morihara(1), Toru Yamashita(1), Koji Abe(2)
1. Okayama University, 2. National Center of Neurology and Psychiatry
Keyword: ischemic stroke, oxidative stress, plasmalogen
Recent studies have consistently demonstrated that early tissue reperfusion confines the ischemic core, leads to a reduction in infarct volume, and restores neurological functions. However, such recanalization may also result in tissue damage known as reperfusion injury. Reactive oxygen species promote the oxidation of cellar DNA, lipids, and proteins, and are overproduced during cerebral ischemia and reperfusion. We previously reported that suppressing oxidative stress by using a free radical scavenger edaravone and a dietary supplement was effective for neuroprotection and improved prognosis in ischemic stroke mice. However, more effective and convenient treatment of ischemic stroke is required. Ascidiacea-derived plasmalogen (aPlas) has both an anti-oxidative molecular structure of a vinyl ether bond at the sn-1 position and the bioactivity of DHA and EPA. In the present study, we examined the possible therapeutic potential of aPlas for 30min transient middle cerebral artery occlusion (tMCAO) mice. Vehicle or aPlas (10 mg/kg/day) were administrated intraperitoneally for two weeks before tMCAO. The mice were continuously treated with vehicle or aPlas, and were sacrificed after five days of reperfusion. aPlas treatment showed a neuroprotective effect on tMCAO mice. aPlas significantly decreased the infarct volume (vehicle: 90.9 ± 21.3 mm3, aPlas: 64.5 ± 23.4 mm3, * p<0.05) and reactive oxygen metabolite levels in serum (d-ROM test; vehicle: 182.0 ± 15.5 CARR U, aPlas: 163.7 ± 12.7 CARR U, * p<0.05). Moreover, aPlas protected the brain against oxidative damage to DNA/RNA and lipid peroxidation, and suppressed the activation of microglia M1, and promoted the regeneration of neurons. These results suggest that aPlas may be a safe and therapeutic candidate against ischemic stroke. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-130
Dynamic quantification of changes in neuronal Na(+) using rapidFLIM with ION NaTRIUM Green-2
*Jan Meyer(1), Niklas J. Gerkau(1), Karl W. Kafitz(1), Matthias Patting(2), Fabian Jolmes(2), Christian Henneberger(3,4,5), Christine R. Rose(1)
1. Institute of Neurobiology, Heinrich Heine University Duesseldorf, 40225 Duesseldorf, Germany, 2. Picoquant, Rudower Chaussee 29, 12489 Berlin, Germany, 3. Institute of Cellular Neurosciences, Medical Faculty, University of Bonn, 53127 Bonn, Germany, 4. German Center for Neurodegenerative Diseases (DZNE), 53127 Bonn, Germany, 5. Institute of Neurology, University College London, London WC1N 3BG, United Kingdom
Keyword: rapidFLIM, Sodium, TRPV4, Ischemia
A fundamental property of animal cells is the maintenance of a low intracellular sodium concentration. This is accomplished by the ubiquitous Na+/K+-ATPase (NKA), which converts ATP into usable energy by keeping up an inwardly directed Na+ gradient across the plasma membrane. In stroke, the oxygen and blood glucose supply is interrupted, leading to a reduction in available intracellular ATP and a breakdown of ion gradients. Fluorescence imaging is an indispensable method for analysis of diverse biological processes enabling e.g. detection of intracellular ions or metabolites. Intensity-based approaches, however, are often prone to artefacts introduced by changes in fluorophore concentrations upon shifts in cellular volume or optical focus drift, which both is the case during stroke. This drawback can be overcome by fluorescence lifetime imaging microscopy (FLIM) based on time-correlated single-photon counting. FLIM is inherently independent of fluorophore concentrations, but often necessitates long photon collection times, resulting in a low temporal resolution. Here, we introduce rapidFLIM featuring drastically decreased dead-time and improved correction algorithms for spatiotemporal imaging of cellular ion signals with low-emission fluorophores. Employing the chemical Na+ indicator ION NaTRIUM Green-2 (ING2), we show that rapidFLIM enables quantitative, dynamic imaging of intracellular Na+ signals in networks of pyramidal neurons of hippocampal mouse brain tissue slices at a full-frame temporal resolution of 2 Hz. Moreover, rapidFLIM of ING2 revealed that brief metabolic inhibition results in unexpectedly large Na+ influx, accompanied by considerable cell swelling. Both were dampened upon the inhibition of TRPV4 channels, identifying this pathway as a hitherto unappreciated driver of neuronal Na+ loading upon metabolic failure. Taken together, rapidFLIM allows the spatiotemporal quantification of ion transients using dim fluorophores at unprecedented speed and independent from changes in cell volume. Moreover, it will enable faster and more sensitive detection of a wide range of dynamic signals with other fluorescent probes, most notably those with intrinsic low photon emission. Supported by the DFG (FOR2795 “Synapses under stress”, Ro2327/13-1). | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-131
脳内出血モデルマウスにおけるSmall Ubiquitin-like Modifier (SUMO)-2/3化修飾の役割
Role of Small Ubiquitin-like Modifier (SUMO)-2/3 conjugation in a mouse model of intracerebral hemorrhage
*木下 慶大(1)、倉内 祐樹(1)、関 貴弘(1)、香月 博志(1)
1. 熊本大学大学院薬物活性学分野
*Keita Kinoshita(1), Yuki Kurauchi(1), Takahiro Seki(1), Hiroshi Katsuki(1)
1. Dept. Chemico-Pharmacol Sci, Grad Sch Pharm, Kumamoto university, Japan
Keyword: INTRACEREBRAL HEMORRHAGE, POST-TRANSLATIONAL PROTEIN MODIFICATION, SUMOYLATION, MICROGLIA
SUMOylation is a kind of post-translational protein modification that occurs through the covalent binding of the Small Ubiquitin-like Modifier (SUMO) peptide to specific target proteins. SUMOylation is involved in normal protein functions, regulating transcription factor transactivation, protein-protein interactions and subcellular localization of certain proteins. Three major SUMO isoforms, SUMO-1, SUMO-2 and SUMO-3, are expressed in the brain, with SUMO-2 and SUMO-3 being very similar each other (sharing 97% sequence homology). In addition to the roles in normal conditions, these SUMO proteins are conjugated by several stimuli including inflammation and oxidant stress. Many studies on the models of brain ischemic stroke have reported that enhanced global SUMOylation observed after stroke plays neuroprotective roles.Intracerebral hemorrhage (ICH) is another type of stroke that is characterized by high mortality and neurological deficits resulting from the formation of hematoma in the brain parenchyma. ICH shows similar inflammatory responses and oxidative stress to those of ischemic stroke, from which neuroprotection could be a therapeutic strategy. In addition, it has been reported that SUMO-1 deficient mice showed more severe neurological dysfunction after ICH induction. However, it remains unclear whether SUMO-2/3 conjugation plays any roles in ICH mouse model. Here, we addressed the relation between SUMO-2/3 conjugation and ICH pathogenesis. ICH was induced in the striatum of male ICR mice by local injection of type VII collagenase. SUMO-2/3 conjugation levels were examined by western blotting at 0, 6, 24, 48 and 72 h after ICH induction. Tissue pathology was examined by immunofluorescence histochemistry at 72 h after ICH induction. SUMO-2/3 conjugation levels were significantly elevated after ICH induction. ICH was accompanied by prominent activation of Iba1-positive microglia/macrophages within the hematoma and GFAP-positive astrocytes in the peri-hematomal region. SUMO-2/3 expression was observed in a subset of these cells at 72 h after ICH induction. In addition, SUMO-2/3 expression was also observed in NeuN-positive neurons in peri-hematomal region at the same time point. These results indicate that SUMO-2/3 conjugation is upregulated in both inflammatory cells and neurons in danger after ICH. The roles of SUMO-2/3 conjugation in regulation of the pathogenic events in ICH will be discussed. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-132
脳梗塞後に海馬歯状回で誘導されるArcadlinが樹状突起スパイン密度に及ぼす影響
Effect of Arcadlin on dendritic spine density in the hippocampal dendtate gyrus after celebral ischemia
*中澤 秀真(1)、井上 耀介(1)、井上 翔太(1)、山口 菜摘(1)、中谷 仁(1)、澤野 俊憲(1)、田中 秀和(1)
1. 立命館大学大学院生命科学研究科
*Nakazawa Shuma(1), Yosuke Inoue(1), Shota Inoue(1), Natsumi Yamaguchi(1), Jin Nakatani(1), Toshinori Sawano(1), Hidekazu Tanaka(1)
1. Ritsumeikan Univ.
Keyword: Arcadlin, Dendritic spine, Hippocampal dentate gyrus, Stroke
脳梗塞は脳内の血流量が減少することによって脳組織が壊死に陥る疾患である。脳梗塞後にはその脳内変化に応じて神経回路のリモデリングが起きることが知られており、その一つとして、樹状突起スパインの変化が知られている。しかしその詳しいメカニズムについては未だ明らかになっていないことも多い。プロトカドヘリン型接着分子であるArcadlinは神経活動依存的に誘導される。Arcadlinはシナプス構造の支持に関与するN-cadherinとホモフィリックに相互作用した後にN-cadherinを伴って細胞内にエンドサイトーシスされ、樹状突起スパイン密度の低下に関与することがわかっている。したがって神経回路リモデリングに応じた樹状突起スパインの変化にArcadlinが関与すると考えているが、脳梗塞後におけるArcadlinの発現とその役割については明らかになっていない。本研究では生後7週齢のC.B-17/Icr-+/Jclマウスに中大脳動脈閉鎖術(MCAO)を施すことで高再現性脳梗塞モデルマウスを作製し、脳梗塞によるArcadlin発現とそれに伴う樹状突起スパインの変化を解析した。In situ hybridization法を用いて、脳梗塞後4時間でのArcadlin発現変動を解析したところ、Arcadlin mRNAが海馬歯状回において強く発現していた。そこで海馬歯状回における樹状突起スパインの形態を蛍光色素ルシファーイエローの細胞内注入によって可視化することで樹状突起スパイン数の変化を検討した。その結果、脳梗塞側海馬歯状回の樹状突起スパイン密度は、偽処置を行なったマウスよりも減少していた。以上の結果より、脳梗塞後における海馬歯状回で強く誘導されたArcadlinが、樹状突起スパイン密度の減少に関与する可能性が示唆された。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-133
脳虚血に対する時間依存的な神経保護メカニズム
Time-dependent neuroprotective mechanism for ischemic stroke
*高橋 弘雄(1)、尾嶋 大喜(1)、坪井 昭夫(2)、山本 融(1)
1. 香川大学分子神経生物学、2. 大阪大学大学院生命機能研究科
*Hiroo Takahashi(1), Daiki Ojima(1), Akio Tsuboi(2), Tohru Yamamoto(1)
1. Dep Mol Neurobiol, Kagawa Univ, Kagawa, Japan, 2. Grad Sch Front Biosci, Osaka Univ, Suita, Japan
Keyword: Ischemia, Npas4, neuroprotection
Ischemic stroke is the second leading cause of death and the most frequent cause of disability in adults worldwide. Stroke initiates complex pathological events in the brain, resulting in cell damage and loss of neurological function. It is important to know the factors that influence neuronal survival or death after stroke. Recently, we reported that mice pre-exposed to an enriched cage (provided with tubes, hiding place, glove etc) for a short period (40 min) acquired resistance to ischemic cell death. The activity-dependent transcription factor Npas4 (neuron PAS domain protein) plays a neuroprotective function in ischemic neurons and is required for this activity-dependent ischemia tolerance (PNAS 118, e2018850118, 2021). The condition of the brain just before stroke has a great impact on ischemic cell death. Interestingly, we found that onset time has a significant effect on cell death after stroke. Infract volume in the cerebral cortex was remarkably reduced when a stroke occurred at night (zeitgeber time (ZT) 15) compared to when a stroke occurred during the day (ZT 6) in mouse stroke models. Npas4 was strongly expressed in the cerebral cortex at night compared to during the day. However, this time-dependent reduction in cell death was also observed in Npas4-deficient mice, suggesting that time-dependent neuroprotection is an Npas4-independent mechanism. Screening for genes with altered expression in ischemic neurons revealed that expression of multiple clock genes was strongly induced under ischemic conditions. We investigated neuroprotective effects of clock genes in ischemic neurons. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-134
Pontine Autosomal Dominant Microangiopathy and Leukoencephalopathyの特異的画像所見であるレーズンパン徴候の診断的意義
“Raisin bread sign” is a specific radiological feature of Pontine Autosomal Dominant Microangiopathy and Leukoencephalopathy
*菊本 舞(1,2)、倉重 毅志(3)、大下 智彦(1,3)、久米 広大(2)、祢津 智久(1)、青木 志郎(1)、越智 一秀(4)、森野 豊之(5)、野村 栄一(6)、山下 拓史(7)、金子 真弓(8)、丸山 博文(1)、川上 秀史(1,2)
1. 広島大学病院脳神経内科、2. 広島大学原爆放射線医科学研究所分子疫学研究分野、3. 国立病院機構呉医療センター脳神経内科、4. 県立広島病院脳神経内科、5. 徳島大学大学院医歯薬学研究部遺伝情報医学分野、6. 広島市立病院機構広島市民病院脳神経内科、7. 広島市立病院機構安佐市民病院脳神経内科、8. 広島市立病院機構安佐市民病院病理診断科
*Mai Kikumoto(1,2), Takashi Kurashige(3), Tomohiko Ohshita(1,3), Kodai Kume(2), Tomohisa Nezu(1), Shiro Aoki(1), Kazuhide Ochi(4), Hiroyuki Morino(5), Eiichi Nomura(6), Hiroshi Yamashita(7), Mayumi Kaneko(8), Hirofumi Maruyama(1), Hideshi Kawakami(1,2)
1. Department of Clinical Neuroscience and Therapeutics, Hiroshima University Graduate School of Biomedical and Health Sciences, Hiroshima, Japan, 2. Department of Epidemiology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, Japan, 3. Department of Neurology, National Hospital Organization Kure Medical Center and Chugoku Cancer Center, Kure, Japan, 4. Department of Neurology, Hiroshima Prefectural Hospital, Hiroshima, Japan, 5. Department of Medical Genetics, Tokushima University Graduate School of Biomedical Sciences, Tokushima, Japan, 6. Department of Neurology, Hiroshima City Hiroshima Citizens Hospital, Hiroshima, Japan, 7. Department of Neurology, Hiroshima City Asa Citizens Hospital, Hiroshima, Japan, 8. Department of Diagnostic Pathology, Hiroshima City Asa Citizens Hospital, Japan
Keyword: familial cerebral small vessel disease, Pontine Autosomal Dominant Microangiopathy and Leukoencephalopathy, magnetic resonance imaging
Objective: Pontine autosomal dominant microangiopathy with leukoencephalopathy (PADMAL) is a hereditary cerebral small vessel disease (cSVD) caused by pathogenic variants in the COL4A1 3’ untranslated region (UTR) and is characterized by multiple ischemic lacunae in the pons. COL4A1 encodes a collagen type IV alpha 1 chain, one of the components of the brain vascular basement membrane. Although several cases of PADMAL have been reported worldwide, including Asia, the suggestive sign for making a diagnosis of PADMAL during their lives has not been established. In this study, we attempted to detect a significant sign for the screening of patients with PADMAL by analyzing the radiological features. Methods: Two patients with cSVD and one unaffected family member in a family (F1) with undetermined familial cSVD were analyzed by whole exome sequencing and Sanger sequencing. We assessed clinicoradiological characteristics of patients in F1. We also clinicopathologically analyzed one of the patients in F1 by performing an autopsy. Subsequently, we screened 40 clinicoradiological features of patients in the cohort of juvenile cerebral vessel disease (CVD) at Hiroshima University Hospital. Sanger sequencing was performed to confirm the variants of the same gene as those in F1. We compared the genetic and radiological features of our cases to those of the previously reported cases. Results: The variant in the COL4A1 3’UTR was detected within the same locus in all patients in F1 but was not detected in the unaffected family member. The locus of their variants was identical to the variant which had been reported in the previous study. Multiple small infarctions and white matter hyperintensities were observed on magnetic resonance imaging (MRI) in both patients, and the appearance of pons with characteristic oval small infarctions resembled “raisin bread”. The autopsy-confirmed patient showed that multiple ischemic lesions in the pons and white matter, which were depicted as oval small infarctions on MRI, were less accompanied by lipohyalinosis and gliosis than ordinary lacuna infarctions. Among the patients in the CVD cohort, we identified two patients who had the same radiological findings as genetically confirmed PADMAL patients in F1. Both patients harbored the variant in the COL4A1 3’ UTR within the same locus as patients in F1. This indicates the significance of this radiological feature in the diagnosis of PADMAL (p=0.0013, Fisher’s exact test). Conclusion: We evaluated radiological features in the pons of PADMAL patients, which were pathologically correlated with pontine multiple oval small infarctions. This radiological feature, for which we coined the name “Raisin Bread Sign”, is highly informative in screening and enables us to perform genetic analysis of COL4A1 in patients with CVD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-135
くも膜下出血後の脳血管攣縮及び神経障害はDAMPs受容体に依存した白血球集積が病因となる。
DAMPs receptor-dependent leukocyte accumulation is critical pathology of cerebral vasospasm/neuronal dysfunction after subarachnoid hemorrhage
*Hiroshi ISHII(1)、Yasuhiro AIDA(2)、Tuyoshi HATTORI(1)、Mika TAKARADA-IEMATA(1)、Mitsutoshi NAKADA(2)、Yasuhiko YAMAMOTO(3)、Osamu HORI(1)
1. 金沢大学医学系神経解剖学、2. 金沢大学医学系脳神経外科学、3. 金沢大学医学系血管分子生物学
*Hiroshi Ishii(1), Yasuhiro AIDA(2), Tuyoshi HATTORI(1), Mika TAKARADA-IEMATA(1), Mitsutoshi NAKADA(2), Yasuhiko YAMAMOTO(3), Osamu HORI(1)
1. Dept Neuroanat, Grad Sch Med, Kanazawa Univ, Kanazawa, Japan, 2. Dept Neurosurgery, Grad Sch Med, Kanazawa Univ, Kanazawa, Japan, 3. Dept Biochem & Mol Vasc Biol, Grad Sch Med, Kanazawa Univ, Kanazawa, Japan
Keyword: subarachnoid hemorrhage, cerebral vasospasm, DAMPs receptor, leukocyte
Purpose: Subarachnoid hemorrhage (SAH) suddenly occurs predominantly by ruptured cerebral aneurysm. The severity of SAH is higher than other types of stroke such as cerebral ischemia and intracranial hemorrhage. It is critical to prevent disability following cerebral vasospasm (CVS) after SAH. Our purpose was to elucidate mechanism underlying CVS and find novel therapeutic target after SAH. Method: In BL6/J background mice, micro-filament, of which tip was cut at right angle, was inserted from left external carotid artery stump to internal carotid artery, and ruptured at the bifurcation of anterior cerebral artery and meddle cerebral artery. Results: In receptor for danger asscoiated molecular patterns (DAMPs) receptor-knockout mice, neurological score and CVS was markedly improved compared with control mice. DAMPs receptor was expressed in endothelial cells of internal carotid artery after SAH. However, vascular specific DAMPs receptor-knockout mice did not show improvement of CVS after SAH. Instead, bone marrow transplantation of wild type mice into DAMPs receptor-knockout mice did not show improvement of these phenotypes compared with control ones into wild mice. Therefore, DAMPs receptor in immune cells contribute to CVS after SAH. Indeed, leukocytes accumulated around cerebral artery after SAH in DAMPs receptor-dependent manner. Conclusion: We newly identified DAMPs receptor as an inducer molecule of CVS/brain injury after SAH. DAMPs receptor in leukocytes is attributed to accumulation around cerebral vessels, which contributes to critical pathology of CVS after SAH. These findings reveal that DAMPs receptor induces leukocyte-mediated CVS after SAH, which suggests that critical pathophysiology of SAH is DAMPs receptor-dependent autoinflammation. This will shed light on the development of essential therapy for SAH patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-136
脳梗塞モデルマウスにおけるSerine protease inhibitor A3N (Serpina3n) の動態
Dynamics of plasma Serine protease inhibitor A3N (Serpina3n) level following focal cerebral ischemia in mice
*江頭 早紀(1)、柏木 寛(1)、森 南海(1)、中野 貴文(1,3)、入江 圭一(1)、細川 雅人(1)、亀谷 富由樹(2)、仲村 佳彦(1)、大坪 俊矢(1)、佐藤 朝光(1)、山下 郁太(1)、佐野 和憲(1)、三島 健一(1)
1. 福岡大学、2. 都医学研、3. 福岡大病院薬
*Saki Egashira(1), Hiroshi Kashiwagi(1), Minami Mori(1), Takafumi Nakano(1,3), Keiichi Irie(1), Masato Hosokawa(1), Fuyuki Kametani(2), Yoshihiko Nakamura(1), Shunya Otsubo(1), Tomomitsu Satho(1), Yuta Yamashita(1), Kazunori Sano(1), Kenichi Mishima(1)
1. Fukuoka Univ., 2. Tokyo metroplitan institute of Medical Sci., 3. Dept. pharm. Fukuoka Univ. Hosp.
Keyword: cerebral infarction
【Introduction】
Treatment with t-PA is effective for patient with cerebral infarction as long as it is received within up to 4.5 hours of the onset. Thus, in the treatment of cerebral infarction, it is important to know the time elapsed after the onset of cerebral infarction in order to provide appropriate treatment. On the other hand, Serpina3n inhibits the protease activity. It is mainly synthesized by hepatocytes and is released in response to early inflammation (Tran M et al., Mol Pharmacol ;99:277–285(2021)). Serpina3n is also known to be increased in muscular dystrophies, which are classified as inflammatory muscle diseases (Tjondrokoesoemo A et al., Hum Mol Genet ;25(6):1192–1202(2016)). Therefore, the amount of Serpina3n is expected to alter in cerebral infarction, but the details are not clear. In this study, we analyzed the dynamics of plasma Serpina3n following cerebral ischemia in mice, and search for new indicators that can help patients with cerebral infarction apply appropriate treatment at the right time.
【Methods】
We used a mouse model of 4-hour middle cerebral arterial occlusion (MCAO) by a nylon monofilament coated with silicone resin. The blood samples were collected at 1 day after MCAO. Proteins in the blood samples were identified by LC-MS/MS. Next, we collected the murine blood samples at 1, 3 and 7 days after MCAO, and measured the concentration of plasma Serpina3n using ELISA kit.
【Results】
The blood samples which were collected at 1 day after MCAO were analyzed by LC-MS/MS. As results, 106 proteins were identified by LC-MS/MS. Among these proteins, 18 types of proteins increased or decreased at 1 day after MCAO. Of these, 11 types of proteins, including Serpina3n, were increased in MCAO mice compared with sham-operated mice. Based on these results, we measured the concentration of plasma Serpina3n at 1, 3 and 7 after MCAO, and found that plasma Serpina3n levels were increased from day 1 to day 7 after MCAO, and peaked at day 3 after MCAO.
【Conclusions】
The results of LC-MS/MS analysis confirmed that 18 proteins altered following cerebral ischemia. It suggested that the alterations of these proteins were associated with progression of disease, and plasma Serpina3n levels could be an indicator to observe progression of cerebral infarction within 3 days after the onset. Measuring Serpina3n levels would be expected to be useful to know the time elapsed after the onset of cerebral infarction in order to provide appropriate treatment. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-137
β - Adrenergic receptor agonist reduce memory interference in APPswe/PS1dE9 mice
*Karunakaran Smitha(1)
*Smitha Karunakaran(1)
1. Centre for Brain Research
Keyword: APPswe/PS1dE9, Memory Interference, Episodic Memory, β-Adrenergic receptor
Earlier studies have demonstrate d that memory dysfunction during early Alzheimer’s disease (AD) is associated with an increased susceptibility to memory interference (MI). In this study, we examined whether previously stored memory representations interfered with n ovel similar input patterns in 2 - month - old APPswe/PS1dE9 (APP/ PS1), mice. APP/PS1 over - expresses the Swedish mutation of amyloid precursor protein together with presen i lin1 deleted in exon 9. A myloid beta (Aβ) plaque deposition is observed at 4 – 6 months of age, not at 2 months providing us with a window to study the earliest behavioural deficits that are independent of Aβ plaque deposition. W e introduced a proactive interference step to the well - established novel object recognition paradigm (NOR), by exposing the mice to multiple objects of different shape, size and colour. Our results demonstrate that NOR is intact in APP/PS1 at 2 months of age. However, introducing a short - term object based priming or interference prior to NOR lead s to recognition memory deficits. This finding indicates that 2 - month - old APP/PS1 is susceptible to proactive MI unlike their Wild Type counterpart who could clearly discriminate between a n ovel and a familiar object. To emulate a real - world scenario, we used Barnes maze, where the mice rely on two - dimensional visual cues of different sh apes to find the escape chamber. We observed an inability in APP/PS1 mice to distinguish extra maze cues that led to delay in learning, an independent evidence for episodic - like memory impairment during spatial learning. O ur findings indicate that, object - based priming can activate its presence during target memory encoding, and thus negate learning in 2 - month - old APP/PS1 mice before amyloidosis onset. Earlier studies have demonstrated that prolonged oral administration of β - adren o receptor (β - AR) agonist (−) - Isoproterenol hydrochloride (IPE) to mice protected them from hippocampal impairment by Aβ oligomers, and enhanced hippocampal synaptic plasticity. We provided the IPE in drinking water for three weeks from P26 till P47. β - AR agonist restored performa nce of APP/PS1 mice to normal levels protecting them from the effects of priming during the object recognition task and ameliorated the deficits noticed in Barnes maze task. These results further indicate that integration or discrimination component of ob ject memory representations relies on and can be modulated by β - AR activation. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-138
ドネペジルとシロスタゾールの低用量併用投与によるアルツハイマー病モデルマウスの記憶障害改善
Combined administration of donepezil and cilostazol ameliorates spatial memory impairment in APP knock-in mouse model for Alzheimer’s disease
*柳井 修一(1)、新崎 智子(1)、斉藤 貴志(2,3)、西道 隆臣(2)、遠藤 昌吾(1)
1. 東京都健康長寿医療センター研究所 老化脳神経科学研究チーム、2. 理化学研究所 脳神経科学研究センター 神経老化制御研究チーム、3. 名古屋市立大学 脳神経科学研究所 認知症科学分野
*Shuichi Yanai(1), Tomoko Arasaki(1), Takashi Saito(2,3), Takaomi Saido(2), Shogo Endo(1)
1. Aging Neuroscience Research Team, Tokyo Metropolitan Institute of Gerontology, 2. Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science, 3. Department of Neurocognitive Science, Institute of Brain Science, Nagoya City University
Keyword: cilostazol, donepezil, Alzheimer’s disease, memory
The combinations of multiple drugs at lower dose are often more effective than using a single drug for treating several diseases. Donepezil, an inhibitor of acetylcholine esterase, is a first-line treatment in patients with mild-to-moderate Alzheimer's disease. The combination strategy, however, has not been established for cognitive impairment related to Alzheimer’s disease. In addition to acetylcholine pathway, cAMP pathway plays essential roles in learning and memory in many species. Recent studies have focused on the cAMP pathway as potential therapeutics for cognitive impairment. Among them, phosphodiesterases (PDEs), which control intracellular cAMP/cGMP concentration, are potential targets for cognitive enhancement. In the present study, we examined the combination of sub-effective doses of donepezil (AchE inhibitor) and cilostazol (PDE3 inhibitor) for memory impairment in APPNL-G-F mice that harbors humanized mutations in the murine Aβ.
The APPNL-G-F mice were divided into 4 groups at the age of 9 months: (1) no drug control group, (2) cilostazol (0.3% in feed) group, (3) donepezil (0.3mg/kg/day in drinking water) group, (4) combined administration group received both cilostazol and donepezil mentioned above. The littermate wild type APPWT mice were also used. After the administration of drugs for 3 months, they were subjected to the behavioral test battery including the open field test and the Morris water maze task.
In the Morris water maze probe test, the no drug control, cilostazol-, and donepezil-administered APPNL-G-F mice performed poorly compared to the littermate WT mice. This demonstrated the impaired spatial memory in 12-month-old APPNL-G-F mice. Furthermore, the drug doses used for single administration of cilostazol and donepezil did not improve spatial memory impairment in APPNL-G-F mice. The combined administration group, however, performed significantly better than the three other groups, allowing them to achieve the level of memory performance similar to the littermate WT mice. In the open field test, the combined administration group showed no significant side effects on locomotor activity and anxiety. The results demonstrated the effectiveness of combined administration at low doses, and this rescue may lead to a novel therapy for treating cognitive impairment in Alzheimer’s disease. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-139
ApoE分泌におけるプレセニリンの必要性
Presenilin is essential for ApoE secretion, a novel role of presenilin involved in Alzheimer’s disease pathogenesis
*鄒 鶤(1)、Sadequl Islam(1)、孫 陽(1)、高 原(1)、中村 知寿(1)、木村 成志(2)、松原 悦郎(2)、春日 健作(3)、池内 健(3)、富田 泰輔(4)、道川 誠(1)
1. 名古屋市立大学大学院医学研究科 神経生化学分野、2. 大分大学医学部 神経内科学講座、3. 新潟大学脳研究所 遺伝子機能解析学分野、4. 東京大学大学院薬学系研究科 機能病態学分野
*Kun Zou(1), Sadequl Islam(1), Yang Sun(1), Yuan Gao(1), Tomohisa Nakamura(1), Noriyuki Kimura(2), Etsuro Matsubara(2), Kensaku Kasuga(3), Takeshi Ikeuchi(3), Taisuke Tomita(4), Makoto Michikawa(1)
1. Dept Biochem, Grad Sch Med, Nagoya City Univ, Nagoya, Japan, 2. Dept Neurol, Sch Med, Oita Univ, Oita, Japan, 3. Dept Mol Genet, Brain Res Inst, Niigata Univ, Niigata, Japan, 4. Dept Neuropathol Neurosci, Faculty Pharm Sci, Univ of Tokyo, Tokyo, Japan
Keyword: Alzheimer's disease, Presenilin, ApoE, Secretion
Alzheimer disease (AD) is a debilitating dementia characterized by progressive memory loss and aggregation of amyloid-β-protein (Aβ) into amyloid plaques in patient brain. Mutations in presenilin (PS) lead to abnormal generation of Aβ, which is the major cause of familial AD (FAD) and apolipoprotein E4 (ApoE4) is the major genetic risk factor for sporadic AD (SAD) onset. However, whether dysfunction of PS is involved in the pathogenesis of SAD is largely unknown. We found that ApoE secretion was completely abolished in PS-deficient cells and markedly decreased by inhibition of γ-secretase activity. Blockade of γ-secretase activity by a γ-secretase inhibitor, DAPT, decreased ApoE secretion, suggesting an important role of γ-secretase activity in ApoE secretion. Reduced ApoE secretion is also observed in nicastrin (NCT) deficient cells with reduced γ-secretase activity. PS deficiency enhanced nuclear translocation of ApoE and binding of ApoE to importin α4, a nuclear-transport receptor. Moreover, expression of PS mutants in PS-deficient cells suppressed the restoration effects on ApoE secretion compared with expression of wild-type PS. Plasma ApoE levels were lower in FAD patients carrying PS1 mutations compared with normal controls. Our findings suggest a novel role of PS contributing to the pathogenesis of SAD by regulating ApoE secretion. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-140
アルツハイマー病におけるミクログリアのカンナビノイド 受容体２型を介した神経炎症調節機序
Stimulation of microglial cannabinoid receptor type II ameliorates cognitive decline in Alzheimer's disease mice by controlling neuroinflammation.
*祖父江 顕(1,2)、小峯 起(1)、遠藤 史人(1)、村山 繁雄(3,4)、斉藤 貴志(5)、西道 隆臣(6)、山中 宏二(1)
1. 名古屋大学環境医学研究所 病態神経科学分野、2. 名古屋大学環境医学研究所 (MIRAIC)-未来の医学研究センター、3. 東京都老人総合研究所高齢者ブレインバンク、4. 大阪大学大学院連合小児発達学研究科附属子どものこころの分子統御機構研究センター ブレインバンク・バイオリソース部門、5. 名古屋市立大学大学院医学研究科脳神経科学研究所、6. 理化学研究所 脳神経科学研究センター
*Akira Sobue(1,2), Okiru Komine(1), Fumito Endo(1), Shigeo Murayama(3,4), Takashi Saito(5), Takaomi Saido(6), Koji Yamanaka(1)
1. Department of Neuroscience and Pathobiology, Research Institute of Environmental Medicine, Nagoya University, 2. Medical Interactive Research and Academia Industry Collaboration Center, Research Institute of Environmental Medicine, Nagoya University, 3. Brain Bank for Aging Research, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology, 4. Brain Bank for Neurodevelopmental, Neurological and Psychiatric Disorders, United Graduate School of Child Development, Osaka University, 5. Department of Neurocognitive Science, Institute of Brain Science, Nagoya City University Graduate School of Medical Sciences, 6. Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science.
Keyword: Alzheimer’s disease , Microglia, neuroinflammation, Astrocyte
Alzheimer’s disease (AD) is the most common form of dementia, characterized by accumulation of amyloid β (Aβ) and phosphorylated Tau. Current therapies for AD are the symptomatic treatment, but do not modify the progression of the disease. On the other hand, neuroinflammation, mediated by activation of glial cells such as astrocytes and microglia, is considered to play an important role in the progression of AD. However, how neuroinflammation contributes to Aβ pathologies in AD remains to be determined. We performed next-generation sequence analysis using RNAs derived from the precuneus of patients with AD, which is selectively vulnerable to amyloid deposition at the early stage, and RNAs derived from microglia isolated from cerebral cortices of 4, 8 and 12-month-old AppNL-G-F mice by using magnetic-activated cell sorting. The expression of cannabinoid type II receptor (CB2) gene was commonly altered in these samples. CB2, mainly expressed in peripheral immune cells and CNS microglia, contributes to modulate inflammation. To elucidate the role of CB2 in the CNS, we analyzed the primary microglia and AppNL-G-F mice using JWH133, a selective CB2 agonist. JWH133 negatively regulated the RNA expression levels of inflammatory cytokines such as TNF-α and CXCL10 in IFN-γ-stimulated microglia in vitro. In addition, chronic oral administration of JWH133 significantly ameliorated the cognitive impairments of AppNL-G-F mice without neuropsychiatric side effects such as hyperlocomotion and anxiety in behavioral tests. JWH133-treated AppNL-G-F mice also showed significant decrease of activated (A1) astrocyte markers and microglial C1q, one of inducers for the A1 astrocyte, compared to vehicle-treated AppNL-G-F mice. JWH133 treatment also inhibited the expression of p-STAT3 (Signal transducer and activator of transcription 3), which is critical for cytokine secretion linked to neuroinflammation, in microglia in the cortex of AppNL-G-F mice. These results suggest that stimulating microglial CB2 ameliorates cognitive dysfunction in AppNL-G-F mice through inducing beneficial neuroinflammation by controlling microglia and astrocyte crosstalk. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-141
神経活動に依存したタウのリン酸化変動
Neuronal activity-dependent alteration of tau phosphorylation
*柳下 聡介(1)、古野 暁子(1)、柴田 恵(1)、橋本 翔子(2)、齊藤 貴志(2,3)、西道 隆臣(2)、荒木 敏之(1)
1. 国立精神・神経医療研究センター、2. 理化学研究所 脳神経科学研究センター 、3. 名古屋市立大学 大学院医学研究科 脳神経科学研究所
*Sosuke Yagishita(1), Akiko Furuno(1), Megumi Shibata(1), Shoko Hashimoto(2), Takashi Saito(2,3), Takaomi C. Saido(2), Toshiyuki Araki(1)
1. National Center of Neurology and Psychiatry, Tokyo, Japan, 2. RIKEN Center for Brain Science, Saitama, Japan, 3. Institute of Brain Science, Nagoya City University Graduate School of Medical Sciences, Aichi, Japan
Keyword: TAU, NEURONAL ACTIVITY, PHOSPHORYLATION, ALZHEIMER DISEASE
Tau is one of microtubule associated proteins, and abnormally phosphorylated tau (pTau) forms neurofibrillary tangles in Alzheimer disease brains. To understand the processes of the neurofibrillary tangle formation, it is required to uncover the mechanisms which alter tau phosphorylation. We have found that intermittent hypoxia treatment might affect neuronal activity resulting in an increase in pTau. We investigated relationship between neuronal activity and pTau using cultured neurons. Neurons were treated with some reagents that could stimulate them, including KCl, AMPA, NMDA, and BDNF. We found that stimuli to neurons cause a decrease in pTau. The extent of the decrease in pTau was inversely-correlated with the extent of an increase in Arc translation. Arc proteins are products of an immediate early gene Arc, with which we estimated the extent of neuronal activity-dependent new protein synthesis. Then, we investigated whether neuronal activity-dependent alteration of pTau could be observed in mice brains. To stimulate neurons in mice brains, we exposed mice to new environment. We found that pTau decreased immediately after exposure to new environment, and remained the level by two hours after the exposure. The level was recovered 24 hours after the exposure. Next, we investigated effect of inhibition of the new protein synthesis on pTau. Mice were treated with reagents which inhibit CREB-dependent protein synthesis. We found that the reagents cause a significant increase in pTau. Finally, we treated with the reagents AppNL-G-F/MAPT double knock in mice, which express human-type tau and show deposition of human-type amyloid β proteins. We found that such reagents cause a significant increase in detergent-insoluble pTau. Our data suggests that pTau is altered dependent on states of neuronal activity-dependent new protein synthesis. In particular, inhibition of new protein synthesis may have important roles in the increase of insoluble pTau. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-142
脳スライス培養におけるAβ含有エクソソームの神経毒性と溶液の流れの作用
Effects of medium flow induced by ciliated cells on neurotoxicity of Aβ containing exosome in the brain organotypic culture
*矢木 圭輝(1)、尾田 空(1)、辰巳 仁史(1)
1. 金沢工業大学大学院工学研究科
*Yoshiki Yagi(1), Sora Oda(1), Hitoshi Tatsumi(1)
1. Kanazawa Institute of Technology
Keyword: Alzheimer’s disease, Aβ, exosome, neurotoxicity
Alzheimer’s disease (AD) is neuropathologically defined by the deposition of intracellular tau aggregates, extracellular amyloid beta (Aβ) deposits, and synapse loss. Aβ is derived by proteolytic processing of amyloid precursor protein (APP), resulting in a peptide predominantly 40 or 42 amino acids in length. Aβ1-42 concentration is higher in the AD brain than cognitively normal elderly individuals, and cerebrospinal fluid (CSF) levels of Aβ1-42 are suggested to be a pathological CSF biomarker signature. The circulation of CSF aids the removal of metabolic waste from the brain, and the clearance rates for Aβ (both Aβ1-42 and Aβ1-40) are impaired in the AD animal compared with controls, supporting the idea that the circulation of CSF the medium flow may interfere with the Aβ neurotoxicity. Recently, exosomes were identified in human CSF and Aβ-containing exosomes are detectable in serum from AD mice and patients, and highly mediate AD. Therefore, studying the action of the flow to the Aβ neurotoxicity mediated by Aβ-containing exosomes is very important. This study focused on the possible mechanism for diminishing the Aβ1-42 neurotoxicity mediated by Aβ containing exosome in the presence of fluid flow. Using brain explant culture and neurons migrated from the explant, we examined that the toxic effects of Aβ under cilia beating induced fluid flow conditions. The neurotoxic effect of Aβ1-42 was detected in neurons on the non-ciliated side, and the neurotoxic action was diminished in neurons exposed to medium flow on the ciliated side. Immunostaining of the Aβ1–42 treated explant culture with the neuronal marker, map-2, and glial marker GFAP showed neurons on the non-cilia side of the explant culture had many blebs, often lost neurites. These neurons had blebs in the soma, and the blebs were positive for Aβ1–42 and GFAP. Immunostaining of the flotillin-2 of Aβ positive particles in culture medium showed the presence of exosomes, which contain Aβ1-42. These results suggest the possibility that the medium flow diminished the accumulation of Aβ-containing exosomes in neurons. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-143
プレセニリン1欠損がACEのAβ変換活性に及ぼす影響
Presenilin 1 deficiency impairs Aβ42-to-Aβ40-converting activity of ACE
*高 原(1)、孫 陽(1)、Islam Sadequl(1)、中村 知寿(1)、富田 泰輔(2)、 鄒 鶤(1)、道川 誠(1)
1. 名古屋市立大学医学研究科、2. 東京大学大学院薬学系研究科
*YUAN GAO(1), Yang Sun(1), Sadequl Islam(1), Tomohisa Nakamura(1), Taisuke Tomita(2), Kun Zou(1), Makoto Michikawa(1)
1. Grad Sch Med Sci, Nagoya City University, Nagoya, Japan, 2. Faculty of Pharmaceutical Sciences, Univ of Tokyo, Tokyo, Japna
Keyword: presenilin, angiotensin-converting enzyme, Alzheimer's disease, Amyloid β-protein
Amyloid β-protein 1-42 (Aβ42) accumulation in brain is the causative factor for Alzheimer's disease (AD). Aβ42 and Aβ40 are the major two species generated from amyloid precursor protein. We have found that angiotensin-converting enzyme (ACE) converts neurotoxic Aβ42 to neuroprotective Aβ40 and this activity is ACE domain and glycosylation dependent. Presenilin 1 (PS1) mutations lead to increased Aβ42/Aβ40 ratio and account for the most of cases of familial AD. However, whether PS1 determines Aβ42/Aβ40 ratio via ACE is largely unknown. Here, we found that purified ACE from PS1 deficient fibroblasts showed significantly reduced Aβ42-to-Aβ40- and angiotensin-converting activities compared with that from wild type fibroblasts. Overexpression of wild type PS1 in PS1 deficient fibroblasts restored Aβ42-to-Aβ40-converting activity of ACE, whereas some PS1 mutants could not. Abnormal glycosylation was also found in ACE purified from PS1 deficient fibroblasts, suggesting that altered glycosylation of ACE results in decreased Aβ42-to-Aβ40-converting activity. Our findings suggest that PS dysfunction or mutations may increase Aβ42/Aβ40 ratio by reducing Aβ42-to-Aβ40-converting activity of ACE. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-144
ApoEアイソフォームによるγセクレターゼ活性の制御
ApoE3 and ApoE4 differentially regulate γ-secretase complex formation and activity
*孫 陽(1)、高 原(1)、Islam Sadequl (1)、中村 知寿(1)、鄒 鶤、道川 誠(1)
1. 名古屋市立大学医学研究科
*YANG SUN(1), Yuan Gao(1), Sadequl Islam(1), Tomohisa Nakamura(1), Kun Zou, Makoto Michikawa(1)
1. Grad Sch Med Sci, Nagoya City University, Nagoya, Japan
Keyword: ApoE, γ-secretase complex
The mechanisms of amyloid accumulation in familial Alzheimer’s disease (FAD) and sporadic AD (SAD) are controversial. Mutations in presenilins (PSEN) found in FAD impair γ-secretase activity and lead to abnormal amyloid β-protein (Aβ) production towards increasing Aβ42/40 ratio. However, SAD is postulated to be caused by the decreased Aβ clearance of apolipoprotein E4 (ApoE4), the strongest risk factor for SAD. Here, by using ApoE3- and ApoE4-knockin (KI) mice and cells, we demonstrated that ApoE3 and ApoE4 bind to γ-secretase complex and isoform-dependently regulate its activity and Aβ production. We found that Aβ40 levels and γ-secretase activity were higher in ApoE knockout mouse brain than wild type mouse brain. ApoE4-KI cells had significant lower Aβ levels and γ-secretase activity, but higher Aβ42/40 ratio compared with ApoE3-KI cells, indicating that ApoE4-KI reduced Aβ production by inhibiting γ-secretase activity. Interestingly, ApoE4 bound more γ-secretase components than ApoE3 and the levels of γ-secretase complex in the brain and cells of ApoE4-KI mice were higher than that of ApoE3-KI mice. Taken together, our findings provide a novel mechanism by which ApoE4 inhibits Aβ production, more preferentially inhibits Aβ40 production and increases Aβ42/40 ratio, via its binding to γ-secretase complex. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-145
タウによるDNA二本鎖損傷修復不全はin vitroにおいてAD病理に関連する
Failure of DNA Double-Strand Break Repair by Tau Mediates AD Pathology in vitro
*浅田 めぐみ(2,1)、植村 健吾(6)、綾木 孝(3)、上村 麻衣子(5)、南山 素三雄(3,7)、引網 亮太(3,7)、守村 敏史(8)、小代 明美(7)、高橋 良輔(3)、植木 孝俊(4)、木下 彩栄(2)、漆谷 真(1,7)
1. 滋賀医科大学 神経難病研究センター、2. 京都大学医学研究科人間健康科学専攻、3. 京都大学医学研究科臨床神経学、4. 名古屋市立大学医学研究科統合解剖学、6. 伊敷病院、7. 滋賀医科大学医学系研究科脳神経内科、8. 滋賀医科大学動物生命科学研究センター
*Megumi utsugi Asada(2,1), Kengo Uemura(6), Takashi Ayaki(3), Maiko Uemura(5), Sumio minamiyama(3,7), Ryota Hikiami(3,7), Toshifumi Morimura(8), Akemi Shodai(7), Ryosuke Takahashi(3), Takatoshi Ueki(4), Ayae Kinoshita(2), Makoto Urushitani(1,7)
1. Molec Neuro Research Center, Grad Sch Med, Univ of Shiga, Otsu, Japan, 2. Dept Human Health Sci, Grad. Sch, Grad Sch Med, Univ of Kyoto, Japan, 3. Dept Neuro, Grad Sch Med, Univ of Kyoto, Japan, 4. Dept of Integrative Anatomy, Grad Sch Med,Univ of Nagoya City, Nagoya, Japan, 5. Center Neurodegenerative Disease Research, Perelman Sch Med, Univ of Pennsylvania, Philadelphia, PA, USA, 6. Ishiki hospital, kagoshima, Japan, 7. Dept Neuro, Grad Sch Med, Univ of Shiga, Otsu, Japan, 8. Research Center for Animal Life Science, Grad Sch Med, Univ of Shiga, Otsu, Japan
Keyword: Tau, DNA double-strand break, microtubule, Alzheimer’s disease
(Background) DNA double-strand break (DSB) is the most severe form of DNA damage and accumulates with age. DNA damage repair and chromosome movement are closely related, while cytoskeleton proteins are responsible for this system. Although evidence indicates that DNA damage is implicated in the pathogenesis of neurodegenerative disease, the exact role of mobility-dependent DNA repair and the microtubule in neurons is unclear. Tau is a microtubule-associated protein, we investigated whether DSB is involved in tau pathologies in Alzheimer’s disease (AD).
(Result) Immunohistochemistry revealed the frequent coexistence of DSB and phosphorylated tau in the cortex of AD patients. We also performed immunofluorescence analysis to identify types of cells vulnerable to DSB damage, astrocytes microglia, endothelial cells and oligodendrocytes, respectively. In the entorhinal cortex and temporal lobe cortex, NeuN - positive neurons co-localized with γH2Ax of DSB marker. In the hippocampus, γH2Ax was frequently colocalized with GFAP. In the cerebral white matter of the temporal lobe,γH2Ax were similarly detected in both AD and the control and were olig2-positive. Microglia and endothelial cells showed almost no co-localization with DSB. Moreover, double immunohistochemistry using AT8 (phosphorylated at S202/T205) and γH2Ax antibodies revealed that cortical cells containing phosphorylated tau (p-tau) occasionally displayed DSB in AD, where p-tau-positive cells were scarce in control. Notably, co-localization of p-tau and γH2Ax was quite rare in the hippocampus of AD brains. In vitro studies using primary cultured neurons showed that non-p-tau accumulated perinuclearly together with the tubulin after DSB induction with etoposide, followed by the conversion to p-tau. Moreover, the knockdown of endogenous mouse tau exacerbated DSB in primary cultured neurons, suggesting the protective role of tau on DNA repair. Interestingly, synergistic exposure of neurons to microtubule disassembly and the DSB strikingly augmented aberrant p-tau aggregation and apoptosis. These data suggest that DSB plays a pivotal role in AD-tau pathology and that the failure of DSB repair leads to tauopathy. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-146
Sirt2はストレプトゾトシン投与によるインスリン欠乏型アルツハイマー病モデルマウス脳内での発現が増加し、ERKの活性化を介してタウのリン酸化を増加させる
Insulin deficiency increases Sirt2 level in streptozotocin-treated Alzheimer’s disease model mice: Sirt2 increases tau phosphorylation through ERK activation
*周 春雨(1)、鄭 且均(1)、Mi-Jeong Kim(2)、渡邉 淳(3)、Mona Abdelhamid(1)、Ferdous Taslima(1)、道川 誠(1)
1. 名古屋市立大学大学院医学研究科・神経生化学、2. 高麗大学校・食品生命工学、3. 国立長寿医療研究センター・研究推進基盤センター・共同利用推進室
*Chunyu Zhou(1), Cha-Gyun Jung(1), Mi-Jeong Kim(2), Atsushi Watanabe(3), Mona Abdelhamid(1), Ferdous Taslima(1), Makoto Michikawa(1)
1. Department of Biochemistry, Nagoya City University Graduate School of Medical Sciences , 2. Department of Food & Biotechnology, Korea University, 3. Laboratory of Research Advancement, Research Institute, National Center for Geriatrics and Gerontology
Keyword: Alzheimer’s disease, Sirt2, tau
Background: Accumulating evidence suggests that insulin deficiency is a risk factor for Alzheimer’s disease (AD); however, the underlying molecular mechanisms are not completely understood. Here, we investigated the effects of insulin deficiency on AD-like pathologies using an insulin-deficient amyloid-β (Aβ) precursor protein (APP) transgenic mouse model (Tg2576 mice).
Methods: Female Tg2576 mice were injected intraperitoneally with streptozotocin (STZ) to induce insulin deficiency, and their body weight, serum glucose levels, and serum insulin levels were evaluated. The levels and deposition of Aβ and the expression of AD pathology-related proteins in the brain were evaluated using immunohistochemistry, enzyme-linked immunosorbent assay, and western blot analysis. To identify the proteins that may affect AD pathologies, cortical proteins from vehicle- and STZ-treated mouse brains were separated by two-dimensional gel electrophoresis (2-DE) and analyzed using liquid chromatography–tandem mass spectrometry (LC–MS/MS).
Results: STZ-treated mice showed exacerbated Aβ accumulation, tau hyperphosphorylation, glial activation, and increased Sirt2 protein levels in the brain, as determined by 2-DE coupled with LC–MS/MS and western blotting. Furthermore, our in vitro experiments revealed that insulin depletion or interleukin-6 treatment increased Sirt2 protein levels and that the overexpression of Sirt2 in both Neuro2a and Neuro2a-P301L cells induced tau hyperphosphorylation through extracellular signal-regulated kinase (ERK)1/2 activation. Conversely, Sirt2 knockdown reversed tau hyperphosphorylation in these cells.
Conclusion: We showed for the first time that Sirt2 is upregulated in the brains of STZ-treated mice and is involved in tau phosphorylation through ERK1/2 activation. Our findings suggest that Sirt2 may be a promising therapeutic target for the treatment of AD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-147
アルツハイマー病における脳の脂質代謝関連因子の変動
Putative role of altered brain lipid metabolism in Alzheimer's disease pathology using postmortem human brain and mice.
*川出 野絵(1)、小峯 起(1)、祖父江 顕(1)、斎藤 貴志(2)、西道 隆臣(3)、山中 宏二(1)
1. 名古屋大学環境医学研究所病態神経科学分野、2. 名古屋市立大学医学系研究科脳神経科学研究所、3. 理化学研究所脳神経科学研究センター
*Noe Kawade(1), Okiru Komine(1), Akira Sobue(1), Takashi Saito(2), Takaomi C Saido(3), Koji Yamanaka(1)
1. Department of Neuroscience and Pathobiology, Research Institute of Environmental Medicine, Nagoya University, Japan, 2. Department of Neurocognitive Science, Institute of Brain Science, Nagoya City University Graduate School of Medical Sciences, Japan, 3. RIKEN Center for Brain Science
Keyword: Alzheimer's disease, lipid metabolism
Obesity is associated with a higher risk for dementia. Several studies reported that obese mice fed high fat diet showed decline in cognitive function. Other human study showed that the white matter volume was decreased in the brain of obese individuals, suggesting that obesity induces brain atrophy. Although obesity could exacerbate brain function in dementia, those mechanisms have not been clarified. In this study, we investigated the changes of lipid metabolism in the brain of Alzheimer’s disease (AD), the most common form of dementia characterized by accumulation of amyloid β (Aβ) and phosphorylated Tau. We performed RNA sequencing of postmortem human brain with early AD pathology. RNAs derived from precuneus, which is selectively vulnerable to amyloid deposition at the early AD stage, were used for this analysis. We found that the several genes related to lipid metabolism, including fatty acid synthesis and lipid uptake, were deregulated in AD brain compared with control brain. Quantitative RT-PCR analyses also identified several deregulated genes linked to the brain lipid metabolism in the brains of AD mice (AppNL-G-F/NL-G-F) carrying App gene with humanized amyloid β sequence, Swedish, Beyreuther/Iberian and Arctic mutations. These results suggested that brain lipid metabolism was altered both in AD patients and AD mice. Based on our results, we speculate that changes of lipid metabolism in AD brain are associated with exacerbation of AD pathology including brain atrophy. We previously reported that oligodendrocyte related genes were decreased in human precuneus with early AD pathology. Therefore, we are in process to perform further analyses of brain lipid metabolism, particularly in oligodendrocyte, using obese AD mice fed high fat diet, and understand the role of obesity in AD pathology through altered brain lipid metabolism. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-148
神経回路形成因子LOTUSとAβ産生酵素BACE1との機能的連関
Functional relationship between the neural circuit formation factor LOTUS and Aβ- producing enzyme BACE1
*上口 碧依(1)、川口 祐生(1)、松林 潤平(1)、竹居 光太郎(1)
1. 横浜市立大学大学院生命医科学研究科
*Aoi Kamiguchi(1), Yuki Kawaguchi(1), Junpei Matsubayashi(1), Kohtaro Takei(1)
1. Grad Sch Med, Univ of Yokohama city, Kanagawa, Japan
Keyword: LOTUS
Amyloid β protein(Aβ) is considered to be a causative factor of Alzheimer's disease. Aβ-producing enzyme is a target of prophylactic and therapeutic drug for Alzheimer's disease. Since β-site APP cleaving enzyme 1(BACE1) is one of the Aβ-producing enzymes, many studies have been conducted with the aim of controlling the activity of BACE1. It is reported that Nogo, an axon growth inhibitor was identified as a BACE1 binding molecule that suppresses the activity of BACE1. Lateral olfactory tract usher substance (LOTUS) discovered in our laboratory binds to Nogo receptor- 1(NgR1) and has a strong antagonistic effect on this receptor. Based on these previous findings, we hypothesized that LOTUS may increase free Nogo by inhibiting the binding of Nogo to NgR1, thereby increasing binding of Nogo to BACE1 and suppressing the activity of BACE1.
In amyloidogenic pathway, BACE1 proteolytically processes APP and produces beta-CTF (C99). In this study, we first examined the expression of CTFs and full-length APP by immunoblotting in cultured mouse cortical neurons and found that CTFs were increased in LOTUS overexpressed cortical neurons. The data suggest that overexpression of LOTUS may regulate BACE1 function. There is a possibility that LOTUS may bind to BACE1 and directly regulate BACE1 function. Therefore, we next examined whether LOTUS binds to BACE1. The binding and pull-down assays showed a possible direct binding of LOTUS to BACE1. Taken together, these findings lead to the possible idea that LOTUS may be involved in the regulation of BACE1 function through binding of LOTUS to NgR1 and/or direct binding of LOTUS to BACE1. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-149
Downregulation of miR-140-5p attenuates memory impairment induced by amyloid-ß oligomer in an animal model possibly through Pin1 regulation
*Pariya Khodabakhsh(1), Maryam Bazrgar(2), Fatemeh Mohagheghi(3), Siavash Parvardeh(1), Abolhassan Ahmadiani(2)
1. Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran, 2. Neuroscience Research Center, Shahid Beheshti University of Medical Science, Tehran, Iran, 3. Institute of Experimental Hematology, Center for Translational Cancer Research (TranslaTUM), School of Medicine, Technical University of Munich, Munich, Germany
Keyword: Alzheimer's disease, Pin1, mir-140-5p, Amyloid-ß oligomer
Pin1 (Peptidylprolyl Cis/Trans Isomerase, NIMA-Interacting 1) has a protective role in the aging-associated neurodegeneration by targeting different phosphorylation sites of tau and the key proteins required to produce Amyloid-β, which are the well-known molecular signatures of Alzheimer’s disease (AD) neuropathology. The direct interaction of miR-140-5p with Pin1 mRNA and its inhibitory role in the protein translation has been identified. The main purpose of this study was to investigate the role of miRNA-140-5p inhibition in promoting Pin1 expression and therapeutic potential of the AntimiR-140-5p in the Aß oligomer (AßO)-induced AD rat model. Spatial learning and memory were assessed in the Morris water maze (MWM). Quantitative RT-PCR, western blot, histological assays were performed on hippocampal samples at various time points after treatments. miRNA-140-5p inhibition enhanced Pin1 mRNA expressions but has little effect on Pin1 protein level. The miRNA-140-5p inhibitor markedly ameliorated spatial learning and memory deficits induced by AßO, and concomitantly suppressed the mRNA expression of inflammatory mediators (Tumour necrosis factor α (TNFα) and Interleukin 1 beta (IL-1β)), and phosphorylation of tau at three key sites (thr231, ser396, and ser404) as well as increased phosphorylated Ser473-Akt. According to our results, Antimir-140-mediated improvement of AβO-induced neuronal injury and memory impairment in rats may provide an appropriate rationale for evaluating miR-140-5p inhibitors as a promising agent for the treatment of AD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-150
末梢Aβはインスリン分泌を抑制する調節因子として働く
Peripheral Aβ acts as a negative modulator of insulin secretion
*富山 貴美(1)、重森 慶子(1)、野村 幸子(1)、梅田 知宙(1)、武田 朱公(2)
1. 大阪市立大学、2. 大阪大学
*Takami Tomiyama(1), Keiko Shigemori(1), Sachiko Nomura(1), Tomohiro Umeda(1), Shuko Takeda(2)
1. Osaka City University, 2. Osaka University
Keyword: Plasma Aβ, Insulin, Alzheimer's disease, Diabetes
Type 2 diabetes mellitus is known to be a risk factor for Alzheimer’s disease (AD), but the underlying mechanisms remain unclear. In AD, the cerebral accumulation of amyloid β (Aβ) triggers a pathological cascade leading to neurodegeneration. Plasma Aβ levels are thought to reflect the brain amyloid pathology and currently used as a diagnostic biomarker of AD. However, amyloid precursor protein and Aβ-generating enzymes, β- and γ-secretases, are widely expressed in various peripheral tissues. Previous reports have shown that glucose and insulin loading cause a transient increase of plasma Aβ in mice and humans. These findings led us to speculate that plasma Aβ is produced from glucose- and insulin-susceptible peripheral tissues to play a role in glucose and insulin metabolism. To test this hypothesis, we investigated the effects of glucose and insulin on Aβ secretion and the effect of Aβ on insulin secretion in vivo, ex vivo, and in vitro. Aβ was found to be secreted from β cells of the pancreas along with insulin upon glucose stimulation. Upon insulin stimulation, Aβ was secreted from cells of insulin-targeted organs, such as adipose tissues, skeletal muscles, and the liver, along with their organokines. Furthermore, Aβ inhibited the glucose-triggered insulin secretion from β cells, slowing down glucose clearance from the blood. These results suggest that peripheral Aβ acts as a negative modulator of insulin secretion. Our findings provide a possible mechanism linking diabetes to AD and call attention to how plasma Aβ levels are used in AD diagnosis. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-151
Diosgeninが脳での軸索再伸長と記憶回復を引き起こす分子メカニズムの解明
Molecular mechanisms for diosgenin-elicited axonal regeneration and memory recovery
*楊 熙蒙(1)、東田 千尋(1)
1. 富山大学 和漢医薬学総合研究所 神経機能学領域
*Ximeng Yang(1), Chihiro Tohda(1)
1. Section of Neuromedical Science, Institute of Natural Medicine, University of Toyama
Keyword: AXONAL REGENERATION, MEMORY RECOVERY
アルツハイマー病（AD）では、脳にAβが沈着することで神経回路網が破綻し、記憶障害が生じる。しかし、記憶障害発症後から破綻した神経回路網を修復し、機能を回復するような治療法は確立されていない。これまでに我々は、山薬（ヤマノイモDioscorea japonicaまたはナガイモD. batatasの根茎）中の成分diosgeninが、初代培養神経細胞においてはAβで萎縮した軸索を再伸長させ、また、ADモデル（5XFAD）マウスに投与すると記憶障害を回復させることを見出してきた。そこで本研究では、diosgenin投与が、5XFADマウス脳内の萎縮した軸索をつながるべき正しい脳部位に再伸長させるかどうかを明らかにし、その現象に関わる分子メカニズムを解明することを目的とした。
海馬から前頭前野への軸索投射を逆行性トレーサーで可視化すると、5XFADマウスでは野生型に比べて投射が減少していた。また、5XFADマウスにdiosgeninを14日間経口投与すると、萎縮した海馬神経細胞の軸索が前頭前野へ再伸長することが示された。これらのマウスの脳切片から、軸索が再伸長した神経細胞をレーザーマイクロダイセクションで採取し、軸索が再伸長しなかった細胞と比較して発現量が変化した遺伝子をDNAマイクロアレイで網羅的に探索した。そのうち、最も発現量が増加した因子A（名称は非公開）について、5XFADマウスの海馬神経細胞に過剰発現すると、マウスの物体認知記憶障害及び空間認知記憶障害が改善し、海馬神経細胞の軸索再伸長が促進された。また、DREADDシステムにより、因子Aを過剰発現した神経細胞特異的に神経活動を抑制すると、因子A過剰発現による記憶改善作用が打ち消された。さらに、神経細胞の軸索上で増加した因子Aは、細胞外のI型collagenと相互作用することで、正しい方向に軸索を再伸長させるという新しい軸索伸長メカニズムも明らかにした。
本研究により、diosgeninが記憶に関わる神経回路において、正しい方向性を持って軸索を再伸長させる薬物であることを見出し、そのメカニズムとして神経細胞中において因子Aが増加することと、その相互作用分子の役割を明らかにした。さらに、因子Aによる軸索再伸長が、記憶回復の直接的かつ十分な要因であることを機能学的に証明した。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-152
筋強直性ジストロフィーにおける大脳誘発電位P1成分は視覚・聴覚・体性感覚ともに振幅増大している
The “P1” components of evoked potentials following auditory, visual and somatosensory stimulation are enhanced in patients with myotonic dystrophy type 1
*諏訪園 秀吾(1)、荒生 弘史(2)、上田 幸彦(3)、前堂 志乃(3)
1. 独立行政法人国立病院機構沖縄病院、2. 大正大学、3. 沖縄国際大学
*Shugo SUWAZONO(1), Hiroshi Arao(2), Yukihiko Ueda(3), Shino Maedou(3)
1. National Hospital Organization Okinawa National Hospital, 2. Taisho University, 3. Okinawa International University
Keyword: event-related potential/evoked potential, myotonic dystrophy, attention, translational research
Myotonic dystrophy type 1, as known as most frequent muscular dystrophy among adults, is associated with well defined gene abnormality (CTG repeat expansion of untranslated area) resulting in dementia. In order to success in translational research for such disease, the logical links among the molecular, the neurophysiological, and the neuropsychological abnormalities need to be established (utilizing surrogate markers), especially in relatively younger adults before the neurodegeneration process impair the therapeutic targets too much. We have reported event-related potentials (Ref.1) utilizing well-defined suitable stimulation paradigms based on neuropsychological data (Ref.2) from more than sixty patients, showing attentional dysfunction mainly.
With this presentation, we will show and discuss the enhanced amplitudes around P1 latencies in three modalities of auditory, visual, and somatosensory, probably related to change in attentional function.
1) Suwazono S, Arao H, Ueda Y, Maedou S. Event-related potentials using the auditory novel paradigm in patients with myotonic dystrophy. J Neurol. 2021 ;268(8):2900-2907. PMID: 33609153
2) Fujino H, Shingaki H, Suwazono S, Ueda Y, Wada C, Nakayama T, Takahashi MP, Imura O, Matsumura T. Cognitive impairment and quality of life in patients with myotonic dystrophy type 1. Muscle Nerve. 2018 ;57(5):742-748. PMID: 29193182 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-153
白質脳症を伴うアミロイド脳血管症における白質の浮腫性変化についての病理学的検討
White matter edematous change in the pre-treatment acute stage of leukoencephalopathy with cerebral amyloid angiopathy
*安田 謙(1)、綾木 孝(1)、Fangzhou Li(2)、桑田 康弘(1)、眞木 崇州(1)、齊ノ内 信(3)、森吉 弘毅(4)、中村 道三(4)、髙橋 良輔(1)
1. 京都大学大学院医学系研究科、2. 浙江大学、3. 新潟大学、4. 京都医療センター
*Ken Yasuda(1), Takashi Ayaki(1), Fangzhou Li(2), Yasuhiro Kuwata(1), Takakuni Maki(1), Makoto Sainouchi(3), Koki Moriyoshi(4), Michikazu Nakamura(4), Ryosuke Takahashi(1)
1. Kyoto University Graduate School of Medicine, Kyoto, Japan, 2. Zhejiang University School of Medicine, Zhejiang, China, 3. Brain Research Institute, Niigata University, Niigata, Japan, 4. National Hospital Organization Kyoto Medical Center, Kyoto, Japan
Keyword: cerebral amyloid angiopathy, CAA, Alzheimer disease
[Objective] Subacute leukoencephalopathy associated with cerebral amyloid angiopathy (CAA) is diagnosed based on biopsy. However, the number of autopsied cases is limited. It is not known whether cases with mild inflammation are attributed to the occasional limitation of the biopsied site. Moreover, little is known about the pathological relationship between edema and vascular lesions in the acute stage of leukoencephalopathy associated with CAA. The purpose of the present study is to elucidate the pathological relationship between edema and vascular lesions in the acute stage of leukoencephalopathy associated with CAA in a autopsied case.
[Methods] A 79-year-old man presented with subacute onset of dementia. Brain MRI disclosed leukoencephalopathy in the posterior lobes with microbleeds. Although clinical manifestation suggested the diagnosis of leukoencephalopathy associated with CAA, he died of sudden rupture of an aneurysm of thoracic aorta two months after the onset of dementia.
[Results] Autopsy demonstrated pathological features of advanced-stage Alzheimer’s disease. Immunostaining with amyloid-β antibody demonstrated CAA in the artery without evidence of capillary CAA. Klüver–Barrera staining showed edematous change predominantly in the white matter of the posterior lobe without ischemic change. Perivascular cuffing was found sparsely, but there was no evidence of angiitis. Pathological findings suggest leukoencephalopathy was caused by the disruption of the blood-brain barrier rather than ischemia. Because the present patient died before immunotherapy, his neuropathological findings could represent the pathomechanism of the acute stage of leukoencephalopathy with CAA.
[Conclusions] CAA leukoencephalopathy could include heterogeneous classifications including amyloid β-related angiitis, CAA-related inflammation, and mild inflammation. Further pathologic studies will be needed to clarify the mechanism of leukoencephalopathy associated with CAA. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-154
Aβ42海馬注入アルツハイマー型認知症モデルマウスにおける海馬依存性記憶の時間帯依存的な想起障害
Time-of-day dependent impairments of memory retrieval in AD model mice micro-infused Amyloid β42 into the dorsal hippocampus.
*ボーグ ジェームス(1)、喜田 聡(1)
1. 東京大学大学院農学系研究科
*James Martin Taihei Borg(1), Satoshi Kida(1)
1. University of Tokyo
Keyword: Alzheimer's Disease, Circadian Rhythm, Long Term Memory, Hippocampus
Alzheimer’s Disease(AD) has been known to show time-dependent declines of cognitive performance. However, mechanisms of this circadian fluctuation of cognitive performance in AD has not been well understood. Interestingly, cognitive performance in humans varies according to time-of-day, with memory retrieval declining in the late afternoon-early evening. By utilizing mice expressing forebrain-specific dominant negative BMAL1(dnBMAL1), we have shown that mice exhibit circadian regulation of memory retrieval, and that hippocampal circadian clock positively regulates memory retrieval (Nature Commun., 2019). To understand the signatures and mechanisms of time-dependent fluctuation of cognitive performance in AD, we have examined time-dependent hippocampal memory performance in AD model mice micro-infused Amyloid β42(Aβ42) into the dorsal hippocampus (Aβ42-injected mice). To do this, altering the combination of the time-of-day the training and testing trials are conducted between ZT4 and ZT10, we uncovered that Aβ42-injected mice show normal memory formation at ZT4 and ZT10 but impaired memory retrieval at ZT10, but not at ZT4. Furthermore, Aβ42-injected mice also
showed similar retrieval impairments of novel object recognition and novel object location memories at ZT10. These results indicate that Aβ42-injected mice exhibit impairment of hippocampal-dependent memory retrieval at ZT10, raising the possibility that these AD model mice show time-of-day dependent fluctuation in cognitive performance, similar to that of AD patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-155
アミロイド前駆体タンパクのプロモーターを標的としたCRISPR/Cas 9によるゲノム編集 −アルツハイマー病への治療応用を目指して−
Targeting the promoter region of amyloid precursor protein by CRISPR/Cas 9 -Toward the therapeutic genome editing for Alzheimer's disease-
*田中 園子(1)、池田 美樹(1)、松村 喜一郎(1)、園生 雅弘(1)、斉藤 史明(1)
1. 帝京大学医学部神経内科
*Sonoko Tanaka(1), Miki Ikeda(1), Kiichiro Matsumura(1), Masahiro Sonoo(1), Fumiaki Saito(1)
1. Sch Med, Teikyo Univ, Tokyo, Japan
Keyword: Alzheimer's disease, amyloid precursor protein, amyloid-β, CRISPR/Cas9
Alzheimer's disease (AD) is the most common cause of dementia and it is estimated that more than 6 million people are affected by this disorder in Japan. The pathological hallmark of AD is the extracellular deposition of amyloid-β (Aβ) in the brain as senile plaques and the formation of intraneural neurofibrillaly tangles consisting of hyperphosphorylated tau. The initial molecular pathogenic step in AD is the excision of Aβ from amyloid precursor protein (APP). In this processing, APP is cleaved first at the N-terminus of Aβ by BACE1 and then at the C-terminus by γ-secretase complex. The cleaved Aβ is secreted into the extracellular space, forms oligomer and exerts cytotoxicity to neurons. The deposition of Aβ is followed sequentially by accumulation of tau, progression of neurodegeneration and finally onset of symptomatic cognitive impairment. Taking these mechanistic insights into consideration, one possible therapeutic target for AD is to reduce the production of Aβ. In this study, we conducted CRISPR/Cas9 genome editing to the APP gene and examined if expression of APP and/or production of Aβ is suppressed by the procedure. First, we designed several guide RNAs targeting the exon 1 and the promoter region of APP gene. These guide RNAs were cloned into a dual-expression plasmid with Cas9 and transfected to HEK293 cells. As expected, quantitative RT-PCR and Western blotting demonstrated that the expression of APP was greatly reduced by the genome editing targeting exon 1. Furthermore, the genome editing against promoter region of APP also decreased the APP production to the comparable degree to the exon 1 targeting. Importantly, Aβ (1-40) and Aβ (1-42) in the culture medium revealed by ELISA analysis were significantly reduced by both of these procedures. Compared to the genome editing of exon 1, the disruption of promoter region is considered to be safe because it doesn't alter the amino acid sequence of APP. This strategy, i.e. targeting the promoter of APP, may contribute to the development of a novel therapeutics for AD in the future. Currently, we are trying to repress the production of Aβ through epigenome editing to the promoter of APP, which is free from double strand break of genomic DNA and thus expected to be much safer than the conventional genome editing. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-156
アルツハイマー病患者iPS由来ニューロンを用いた疾患モデル構築の有用性評価
Evaluation of utility of Alzheimer’s disease model using patient iPSC-derived neurons
*荻原 京加(1)、邉見 奈津子(1)、瀬尾 学(2)、東 基記(1)
1. 株式会社リコー、2. Elixirgen Scientific, Inc.
*Miyaka Ogihara(1), Natsuko Hemmi(1), Manabu Seo(2), Motoki Azuma(1)
1. RICOH Company, Ltd., Kanagawa, Japan, 2. Elixirgen Scientific, Inc., Baltimore, US
Keyword: Alzheimer's disease, iPSC-derived neuron, Drug discovery, Phenotypic screening
Alzheimer’s disease is the most common form of dementia. Despite the increasing prevalence of this disease in an aging society, there are still no effective treatments. Since no animal models fully reproduce the pathology of Alzheimer’s disease and species differences also exist in drug action. Therefore, patient-derived iPSCs have been utilized as one of human in vitro models for drug discovery research. The pathology of Alzheimer's disease is characterized by extracellular accumulation of amyloid plaques, and intracellular neurofibrillary tangles of phosphorylated Tau. Although it has been reported that accumulation of Aβ and⁄or phosphorylated Tau are also observed in patient iPSCs-derived neurons, it usually takes several months for differentiation and maturation of iPSCs into neurons. Thus, we evaluated the phenotype of patient-derived neurons using Quick-Neuron™, which differentiated iPS cells into neurons rapidly, in order to develop a quick and easy disease model with iPS cells for drug discovery research.
Quick-Neuron™ from sporadic Alzheimer's disease patient and healthy donor were plated and cultured for one to eight weeks. Cell lysates and culture media were collected at each time point, and the accumulation of Aβ in the culture media and intracellular total Tau and phosphorylated Tau in the cell lysates were evaluated by ELISA. After six weeks culture, Aβ40 and Aβ42 in the culture media were significantly accumulated in neurons derived from Alzheimer's disease iPSCs. Tau accumulation and Tau phosphorylation were significantly induced in neurons derived from Alzheimer's disease iPSCs after four weeks culture. We also showed that β-secretase inhibitors, Aβ production inhibitors, significantly reduced the accumulation of Aβ40 and Aβ42 in patient-derived neurons.
These results imply that patient-derived Quick-Neuron™ can reproduce the clinical symptoms of Alzheimer's disease in four to six weeks, and that can be a useful model for Alzheimer's disease. Furthermore, the expected response to β-secretase inhibitors, one of the important targets of the drugs for Alzheimer's disease, suggested that this model can be applied to drug discovery screening. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-157
アルツハイマー病モデルマウスにおける認知機能障害および神経炎症に対するフマル酸ジメチルの影響
Beneficial effects of Dimethyl Fumarate on the cognitive impairment and neuroinflammation in Alzheimer's disease mice
*Wang Ting(1)、祖父江 顕(1,2)、小峯 起(1)、渡邊 征爾(1)、斉藤 貴志(3)、西道 隆臣(4)、山中 宏二(1)
1. 名古屋大学環境医学研究所 病態神経科学分野、2. 名古屋大学環境医学研究所附属MIRAIC-未来の医学研究センター、3. 名古屋市立大学脳神経科学研究所・認知症科学分野、4. 理化学研究所脳神経科学研究センター 神経老化制御研究チーム
*Ting Wang(1), Akira Sobue(1,2), Okiru Komine(1), Seiji Watanabe(1), Takashi Saito(3), Takaomi Saido(4), Koji Yamanaka(1)
1. Nagoya universityDepartment of Neuroscience and Pathobiology, Research InstiDepartment of Neuroscience and Pathobiology, Research Institute of Environmental Medicine, Nagoya University,Japan, 2. Medical Interactive Research and Academia Industry Collaboration Center, Research Institute of Environmental Medicine, Nagoya University,Japan, 3. Department of Neurocognitive Science, Institute of Brain Science, Nagoya City University Graduate School of Medical Sciences Japan, 4. Laboratory for Proteolytic Neuroscience, RIKEN Center for Brain Science Japan
Keyword: Alzheimer's disease, neuroinflammation, Nrf2, Astrocyte
Neuroinflammation, mediated by the activation of microglia and astrocytes in the central nervous system and subsequent production of proinflammatory molecules, plays a significant role in the pathologies of Alzheimer’s disease (AD). AD is the most common form of dementia, characterized by accumulation of amyloid β and phosphorylated Tau. Previous studies reported nuclear factor erythroid 2-related factor 2 (Nfe2l2 or Nrf2) attenuated inflammation in neurodegenerative diseases, however, the detailed mechanism is unclear. Our RNA sequence analysis on isolated-glial cells from the cortices of AppNL-G-F/NL-G-F (App-KI) mice identified deregulation of Nrf2-regulated genes and neuroinflammatory factors in astrocytes of App-KI mice. To elucidate the detailed mechanism of activating Nrf2 pathway as the treatment for AD through regulating neuroinflammation, we examined the role of Nrf2 in vitro and vivo by using Dimethyl Fumarate (DMF), a clinically available drug to activate Nrf2 pathway. In vitro, we used three humoral factors, IL-1α, TNF-α, and C1q together to induce activated (A1) astrocytes, and indicated the regulatory role of DMF on A1/A2 astrocytic alteration through the upregulation of Nrf2 pathway. DMF significantly downregulated the mRNA expression of A1 genes such as H2-d, H2-t23, and Gbp2, and enhanced the mRNA expression of Nrf2 and its downstream genes in primary astrocytes. Meanwhile, the levels of S100a10, Cd109, and Clcf1 were significantly increased after DMF exposure. Moreover, chronic oral administration of DMF significantly ameliorated the cognitive impairments of App-KI mice using novel object recognition test, while the same treatment had negligible effects on normal cognitive function in wild-type mice. In MACS-isolated-astrocytes of DMF treated-App-KI mice, the expression of H2-d was significantly decreased and upregulated the expression of Osgin1. DMF treatment also reduced amyloid-β deposition in the cortices of App-KI mice. These results suggest that DMF ameliorates neuroinflammation and cognitive impairments of App-KI mice by suppressing reactive astrocytes through activating Nrf2 pathway. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-158
Autobiographical Episodic Memory of Chinese-Speaking Seniors with Alzheimer’s Disease
*Yi-hsiu Lai(1), Yu-te Lin(2)
1. National University of Kaohsiung, Taiwan, 2. Kaohsiung Veterans General Hospital, Taiwan
Keyword: Alzheimer’s disease, Mandarin Chinese, Autobiographical episodic memory
This study examined autobiographical episodic memory of Chinese-speaking seniors with Alzheimer’s disease (AD). Episodic autobiographical memory is regarded as a key characteristic of an individual’s life experience. It is widely reported that aging affected episodic autobiographical memory negatively (Craik and Grady, 2002; Craik & Salthouse, 2007; Moscovitch & Winocur, 1992; Sagar & Corkin, 1991; Seidl et al., 2011; St-Laurent et al., 2011). Issues concerning task effects and period effects on the AD autobiographical episodic memory, nonetheless, have been less addressed. Little attention has been drawn to autobiographical episodic memory of Chinese-speaking AD seniors. In this study, autobiographical episodic memory of thirteen Chinese-speaking AD persons were collected and compared with twenty healthy controls. According to the NINCDS-ADRDA criteria (McKhann et al., 1984), the AD participants were clinically-diagnosed as AD, not from other types of dementia. Linguistic variables included the period effects (i.e., three broad time-periods: childhood, early youth and recent life) and the task effects (unguided episodic memory vs. guided episodic memory). Results indicated a number of significant findings regarding AD autobiographical episodic memory. We also elaborated how periods effects and task effects differentiated Chinese-speaking AD seniors from healthy seniors. In conclusion, the current investigation contributed the impaired autobiographical episodic memory of Chinese-speaking AD seniors, and provided some suggestions for future research.
References
Craik, F I.M., & Grady, C.L. (2002). Aging, memory, and frontal lobe functioning. In D. T. Stuss & R. Knight (Eds.), Principles of frontal lobe function (pp. 528-540). New York: Oxford University Press. Craik, F. I. M. & Salthouse, T. A. (2007) (Eds.), The handbook of aging and cognition (pp. 315-372). Hillsdale, NJ: Erlbaum. McKhann, G., Drachman D., Folstein M., Katzman R., Price D., & Stadlan Emanuel M. (1984). Clinical diagnosis of Alzheimer's disease: report of the NINCDS-ADRDA Work Group under the auspices of the Department of Health and Human Services Task Force on Alzheimer's disease. Neurology, 34, 939-944. Moscovitch, M., & Winocur, G. (1992). The neuropsychology of memory and aging. In T.A. Sagar, H.J., Edith, V. S., & Corkin, S. (1991). Autobiographical memory in normal ageing and dementia. Behavioural Neurology, 4(4), 235-248. Seidl, U., Lueken, U., Thomann, P.A., Geider, J., & Schroder, J. (2011). Autobiographical memory deficits in Alzheimer’s disease. Journal of Alzheimers Disease, 27, 567-574. St-Laurent, M., Abdi, H., Burianova, H., & Grady, C.L. (2011). Influence of aging on the neural correlates of autobiographical, episodic, and semantic memory retrieval. Journal of Cognitive Neuroscience, 23(12), 4150-4163. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-159
次世代型タウオパチーモデルマウスの開発
Development of next-generation tauopathy mouse model
*綿村 直人(1)、Foiani Martha(2)、Duff Karen(2)、西道 隆臣(1)
1. 理化学研究所、2. University College London, DRI
*Naoto Watamura(1), Martha Foiani(2), Karen Duff(2), Takaomi Saido(1)
1. RIKEN, 2. University College London, DRI
Keyword: Tauopathy, Genome editing, Base editor
Aim: Tauopathies are a group of pathological disorders characterized by the accumulation of filamentous phosphorylated tau in the brain. To study the pathogenesis of the disease and to understand the neurotoxic effects of tau in dementia, we aimed to develop mouse models that reproducibly display specific tau pathology without artifacts because the current tauopathy models depend on the overexpression paradigm. The purpose of this study is to develop new tauopathy mouse models by knock-in strategy. Methods: Previously we generated human MAPT knock-in (hTau-KI) mice, in which the entire Mapt gene was humanized. The hTau-KI mice express all six isoforms (Hashimoto et al.,2019; Saito et al., 2019). In this study, several tauopathy-causing mutations were introduced into hTau-KI mice by Base Editor, a new CRISPR/Cas9-based genome editing technology. Results: We obtained several tauopathy mouse models (hTau-KI P301L, hTau-KI P301S, hTau-KI P301V, hTau-KI Intron10+3 G>A, hTau-KI P301L; Intron10+3 G>A, hTau-KI P301S; Intron10+3 G>A, hTau-KI S305N; Intron10+3 G>A). Immunohistochemical analyses showed phospho-tau positive signals in the entorhinal cortex and other areas in some of these lines. In addition, biochemical analyses demonstrated that the intronic mutation altered the expression patterns of 3 and 4 repeat tau. Finally, we crossbred these mutant hTau-KI mice with App knock-in mice and found that amyloid pathology accelerated the tau pathology in the brain. Conclusions: We successfully introduced tauopathy-causing mutations in hTau-KI mice. Consequently, we identified the effects of these MAPT mutations on the pathological and cognitive parameters. We expect these models to contribute to the research community for better understanding the mechanisms of tauopathies. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-160
持続的な高体温は認知機能とアルツハイマー病に関連する病態を悪化させる
Sustained high body temperature exacerbates cognitive function and Alzheimer’s disease-related pathologies
*鄭 且均(1)、加藤 玲子(2)、周 春雨(1)、Abdelhamid Mona(1)、山下 均(2)、道川 誠(1)
1. 名古屋市立大学大学院医学研究科、2. 中部大学
*Cha-Gyun Jung(1), Reiko Kato(2), Chuny Zhou(1), Mona Abdelhamid(1), Hitoshi Yamashita(2), Makoto Michikawa(1)
1. Nagoya City University Graduate School of Medical Sciences, 2. Chubu University
Keyword: Alzheimer’s disease, Sustained high body temperature, Memory impairment, Tau
Global warming is a serious public health threat to people worldwide. High body temperature is one of the important risk factors for Alzheimer’s disease (AD), and the body temperature of AD patients has been found to be significantly higher than that of elderly control subjects. However, the effects of high body temperature on cognitive function and AD pathologies have not been completely elucidated. We report here that amyloid-β precursor protein (APP) transgenic mouse (Tg2576) housed at a high ambient temperature of 30℃ for 13 months showed an increase in the body temperature, which is accompanied by memory impairment and an enhancement of amyloid-β peptides (Aβ) generation through the upregulation of β-site APP cleaving enzyme 1 (BACE1) level and decrease in the level of an Aβ-degrading enzyme, neprilysin (NEP) in the brain, compared with those of Tg2576 mice at 23℃. High body temperature also increased the levels of heat shock proteins (HSPs), stress-stimulated kinases such as JNK, and total tau, leading to the enhancement of tau phosphorylation in the brain of mice at 30℃. Taken together, our findings suggest that high body temperature exacerbates cognitive function and AD pathologies, which provides a mechanistic insight for its prevention. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-161
D-セリン依存的なアルツハイマー病の病態進行
D-Serine dependent pathophysiological progression of Alzheimer’s disease
*倪 献策(1,2)、井上 蘭(1,3)、齊藤 貴志(6)、西道 隆臣(6)、高雄 啓三(2,3,4,5)、森 寿(1,2,3,4)
1. 富山大学 医学部 分子神経科学研究講座、2. 富山大学 生命融合科学教育部、3. 富山大学 アイドリング脳研究センター、4. 富山大学 未病研究センター、5. 富山大学 医学部 行動生理学講座、6. 理化学研究所 脳神経科学研究センター（理研CBS）
*Xiance Ni(1,2), Ran Inoue(1,3), Takashi Saito(6), Takaomi Saido(6), Keizo Takao(2,3,4,5), Hisashi Mori(1,2,3,4)
1. Department of Molecular Neuroscience, Faculty of Medicine, University of Toyama, 2. Graduate School of innovative Life Science, University of Toyama , 3. Research Center for Iding Brain Science (RCIBS), 4. Research center for Pre-Disease Science, University of Toyama, 5. Department of Behavioral Physiology, Faculty of Medicine, University of Toyama, 6. Laboratory for Proteolytic Neuroscience, Brain Science Institute, RIKEN, Wako, Saitama, Japan
Keyword: Alzheimer’s disease, D-serine, serine racemase, NMDAR
Alzheimer’s disease (AD) is the most common form of dementia, with a complex pathophysiological progression. The essential pathological changes include extracellular accumulation of amyloid β (Aβ), intracellular neurofibrillary tangles, and cerebral atrophy due to degenerative neuronal death. The N-methyl-D-aspartate receptor (NMDAR) plays pivotal roles in excitatory neurotransmission and synaptic plasticity, in turn, related to cognitive ability. Neurodegenerative processes in AD are also associated with excitotoxicity induced by NMDAR overactivation. Therefore, NMDARs are considered to highly participate in the pathogenesis of AD. D-Serine, as the major endogenous co-agonist of NMDAR, is produced by serine racemase (SRR) in mammalian brains. Further, it has been reported that D-serine level and SRR expression pattern were altered in AD models and Aβ induced neuronal damage was suppressed in SRR knockout (KO) mice, suggesting that D-serine is associated with the development of AD pathophysiology. However, the role of D-serine in AD has not been entirely elucidated. Using a 3rd generation AD model, AppNL-G-F/NL-G-F (APPKI) mouse, we examined characteristic pathophysiology including Aβ deposition and glial activation, and compared SRR levels between APPKI and control mice. Glial fibrillary acid protein (Gfap) and ionized calcium-binding adapter molecule 1 (Iba1) were used as markers for astrocytes and microglia, respectively. We observed age-dependent increases in Aβ burden and gliosis from 6 to 12-months old as previously reported, and we found that SRR level was decreased in the cerebral cortex of APPKI mice at 12-month old compared to age-matched controls, but the neuronal density was unaltered. Three-month supplements of D-serine in APPKI mice from 9 to 12-month old led to neuronal loss in the hippocampal CA1 region, which suggests that D-serine is required for NMDAR-mediated excitotoxicity in AD. Besides, downregulation of SRR in APPKI mice may display a self-protective role against excitotoxicity. To further determine the role of D-serine in AD pathology, we crossed APPKI mice with SRRKO mice to generate SRR gene-deleted APPKI (APPKI; SRRKO) mice. APPKI; SRRKO mice showed lower Gfap levels and decreased Iba1-positive cells at 9-month old in comparison to APPKI mice. These results indicate that D-serine is tightly involved in the pathological changes of AD, such as excitatory neurotoxicity and AD-related inflammatory responses. Moreover, inhibiting D-serine signaling may be an effective method to alleviate AD pathology. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-162
タウオパチーモデルマウスにおけるタウを介する神経変性に対するアンセリンの効果に関する研究
The effects of anserine treatment on tau-mediated neurodegeneration in a mouse model of tauopathy
*雷 晨旭(1)、久恒 辰博(1)
1. 東京大学大学院 新領域創成科学研究科
*CHENXU LEI(1), Tatsuhiro Hisatsune(1)
1. Graduate School of Frontier Sciences, The University of Tokyo
Keyword: Alzheimer's disease, Tau pathology, Neuroinflammation, Anserine
Alzheimer’s disease (AD) is a progressive neurodegenerative disease characterized by the deposition of amyloid-β (Aβ) and neurofibrillary tangles (NFTs) caused by abnormal phosphorylated tau aggregation. Compared with senile plaques caused by Aβ deposition, NFTs in AD brain has a closer correlation with clinical signs, which are closely related to synapse loss, neuroinflammation and neurodegeneration (Arriagada et al, 1992). Therefore, targeting of tau pathology is regarded as a promising strategy for AD treatment.
Neuroinflammation in the central nervous system (CNS) is closely related to the progression of tau pathology, which is mediated by the activation of microglia and astrocytes. The inhibiting of neuroinflammation has been suggested to reduce tau pathology and tau-mediated neurodegeneration (Laurent et al, 2018). Anserine, which is a methylated form of carnosine, has been reported to have an ability to reduce neuroinflammation and protect cognitive decline (Kaneko et al, 2017), but the connections between the reduction of neuroinflammation and the improvement of cognitive function has not been fully clarified. In this study, we proposed a hypothesis that anserine may reduce neurodegeneration by reducing neuroinflammation to protect cognitive function.
Human APOE4 KI/P301S Tau mice (TE4) were used to study the effects of anserine on neurodegeneration and an 8-weeks anserine treatment (10mg/mouse per day) was performed from 6 to 7 months of age. TE4 mice showed a significant decline in cognitive function at 8 to 9 months of age in Y-maze and anserine treatment improved the cognitive function of TE4 mice (P<0.05).
Then we studied the effects of anserine on tau pathology and neurodegeneration. TE4 mice were found to show a significant neuronal loss in CA1 area of hippocampus. After anserine treatment, neuronal loss was significantly decreased. Moreover, the number of NFTs in CA1 was decreased in TE4 (A) mice (with anserine treatment). These results indicated that anserine treatment can reduce tau-mediated neurodegeneration.
To further study whether the reduction of neurodegeneration was related with inhibited neuroinflammation, we performed GFAP and Iba1 staining. Anserine treatment did not reduce the number of Iba1+ cells but significantly reduced the number of GFAP+ cells in TE4 mice (P<0.01). An interesting thing was that the number of GFAP+ cells was related to the neurodegeneration levels in CA1 (thickness and NFT number). Further studies suggested that this may be related to the decreased expression of complement C3 (P<0.05), which is secreted by activated astrocytes.
In general, our findings suggested anserine may inhibit the activation of astrocytes to reduce tau-medicated neurodegeneration, suggesting that anserine has promise as a therapeutic drug for the prevention and treatment of AD in the future. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-163
認知症における可逆的記憶障害の定量的・継時的解析
Quantitative and successive analysis of reversible memory impairment in dementia
*澁木 克栄(1,3,4,5)、田中 晋(2)、森田 昌宏(2)、田中 政春(2)、染矢 俊幸(1)、田中 弘(2)
1. 新潟大学医歯学総合研究科、2. 三島病院、3. 柏崎総合医療センター、4. 新潟県立精神医療センター、5. 新潟大学脳研究所
*Katsuei Shibuki(1,3,4,5), Shin Tanaka(2), Masahiro Morita(2), Masaharu Tanaka(2), Toshiyuki Someya(1), Ko Tanaka(2)
1. Grad Sch Med Dent, Niigata Univ, Niigata, Japan, 2. Mishima Hospital, Niigata, Japan, 3. Kashiwazaki General Hospital and Medical Center, Niigata Japan, 4. Niigata Psychiatric Center, Niigata, Japan, 5. Brain Res Inst, Niigata Univ, Niigata, Japan
Keyword: Dementia, Memory impairment, Memory test
Most cases of dementia are caused by irreversible neurodegeneration, so that memory impairment in dementia has been considered to be irreversible. However, the memory impairment may include reversible components derived from functional depression of synaptic function as well as irreversible components resulting from neurodegeneration. If so, appropriate training of the brain functions could reverse a part of the memory impairment. To test this possibility, it is necessary to measure the memory capacity of dementia patients quantitatively and successively. We developed a PC-based game for quantitative and repeated measurement of visual memory capacity, and asked dementia patients to play the game, and analyzed the changes in game performance or visual memory capacity. Of the pictograms open in the internet, 5000 images were selected that were easily discriminated by the elderly people. A sample image in each trial was randomly selected from the 5000 images, and demonstrated to a dementia patient. The patient was asked to perform a simple calculation for the following period of 10 seconds to prevent short-term memory. Then the patient was asked to select one correct image, which was shown before the calculation, among five different images. The memory capacity was judged by whether the correct sample image could be selected. The level of difficulty could be adjusted by changing the presentation time of the sample image, and a total of 30 trials were conducted per day, and the percentages of correct trials in the 60 trials for 2 days were used as an index of the memory capacity. Three of the five patients with mild dementia, judged from the MMSE and HDS-R scores, showed a significant increase in game performance within 4 sessions. The increase in game performance continued slowly thereafter, and one of them showed a significant increase in game performance again after switching to a more challenging task using two sample images. Two patients with moderate dementia, however, showed no increase in game performance or no significant memory for this task. These results suggest that memory impairment in dementia may contain reversible and treatable components presumably resulting from reversible changes in synaptic function. Naturally, however, future studies are necessary to determine whether improved game performance is associated with improved cognitive function in dementia patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-164
Rosa26ノックイン型タウ過剰発現マウスを用いたタウオパチーにおける神経炎症応答解析
A novel Rosa26-Knockin Tau expressing mouse model exhibits neuroinflammatory responses during tauopathy
*矢内 凜(1)、南久松 丈晴(1)、下條 雅文(1)、高堂 裕平(1)、佐原 成彦(1)、樋口 真人(1)
1. 国立研究開発法人量子科学技術研究開発機構
*Rin Yanai(1), Takeharu Minamihisamatsu(1), Masafumi Shimojo(1), Yuhei Takado(1), Naruhiko Sahara(1), Makoto Higuchi(1)
1. National Institutes for Quantum Science and Technology
Keyword: Tau, Tauopathy model, neuroinflammation, microglia
Animal modeling of neurodegenerative disorders is essential for understanding causal mechanisms of neurodegeneration and development of therapeutic interventions. Tauopathy is one of major pathological hallmarks of neurodegenerative diseases. Since several mutations in tau gene were identified in FTDP-17, overexpression of familial mutant tau by transgenesis has been utilized to drive tauopathy and disease-relevant phenotypes. However, recent studies revealed that the transgenes disrupt the coding sequence of endogenous genes resulting deletions and/or structural variations at the insertion site (Goodwin et al. 2019). In this study, we utilized the knockin (KI) strategy to insert P301L mutated 1N4R human tau cDNA including the tetracycline response element (TRE) promoter into the Rosa26 safe harbor locus. To generate a human tau expressing mouse, the KI mouse was cross-bred with CaMKII-tTA transgenic mice under C57BL/6J background. The resultant mouse expresses P301L human tau in the forebrain with 4-fold of mouse tau in the absence of dentate granule neuronal degeneration, which was appeared in CaMKII-tTA transgenic mice under 129/Sv background (Han et al. 2012). Immunohistochemical and biochemical analyses revealed that tau pathologies were observed and extended in cerebral cortices and hippocampi of homozygous mice for P301L human tau gene from 12 months of age. In parallel with pathological tau progression, Iba1-positive microglia and GFAP-positive astrocytes were increased. As counterparts of microglial activation, homeostatic microglial markers, P2RY12 and Tmem119 were decreased in AT8 (p-tau specific antibody)-positive hippocampal regions. Significant correlations between AT8 signal and levels of microglial markers were observed in hippocampal regions of 12-15-month-old homozygous tau-KI mice. We further confirmed microglial activation in the hippocampus by immuno-staining of Axl, which is one of markers of disease associated microglia. In summary, the present mouse model shows spaciotemporal progressions of tau pathology and glial activation. This model will provide useful information of a linkage between p-tau-bearing neurons and activated microglia. Further examinations are ongoing for the elucidation of tau-induced neurotoxicity in association with neuroinflammation. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-165
テストステロンはグリア発現アンドロゲン受容体刺激によりアルツハイマー病マウスにおける神経炎症を制御する
Testosterone regulates the neuroinflammation in Alzheimer’s disease mice by stimulating glial androgen receptors
*前川 華澄(1)、祖父江 顕(1)、小峯 起(1)、斉藤 貴志(2)、西道 隆臣(3)、山中 宏二(1)
1. 名古屋大学 環境医学研究所 病態神経科学分野、2. 名古屋市立大学 大学院医学研究科 脳神経科学研究所 認知症科学分野、3. 理化学研究所 脳神経科学 研究センター 神経老化制御研究チーム
*Kasumi Maekawa(1), Akira Sobue(1), Okiru Komine(1), Takashi Saito(2), Takaomi Saido(3), Koji Yamanaka(1)
1. Dept of Neurosci & Pathobiol, RIEM, Nagoya Univ, Aichi, Japan, 2. Dept Neurocog Sci, Inst of Brain Sci, Nagoya City Univ Grad Sch of Med Sci, Aichi, Japan, 3. Lab for Proteolytic Neurosci, RIKEN CBS, Saitama, Japan
Keyword: Alzheimer's disease, Neuroinflammation, Microglia, Astrocytes
Neuropathology of Alzheimer’s disease (AD) is characterized by the accumulation of amyloid β (Aβ) and phosphorylated Tau. Microglia and astrocytes play critical roles in the AD pathologies such as the accumulation of Aβ and subsequent neuroinflammation. Our result of RNA sequencing has shown that the expression of androgen receptor (AR) was decreased in the human precuneus with early AD pathology and isolated microglia and astrocytes from the cerebral cortices of AppNL-G-F/NL-G-F (App-KI) mice, the new generation AD model. Although previous studies have demonstrated that the correlation between low testosterone level and the risk of AD, the detailed roles of testosterone in AD pathology and the functions of glial AR remain unclear. Therefore, we investigated the effect of testosterone through glial AR on AD pathology using castrated-App-KI male mice.
To examine whether the deprivation of testosterone worsens the AD pathology in App-KI mice, we castrated App-KI male mice at 4 months of age and have checked that plasma testosterone level in castrated-App-KI male mice was lower than the one in sham-App-KI male mice. We isolated microglia and astrocytes from the cerebral cortices by magnetic activated cell sorting (MACS) at 18 months of age for RT-qPCR. In isolated-microglia of aged castrated-App-KI mice, mRNA levels of pro-inflammatory molecules (Cxcl10 and Ccl5) tended to be increased. Meanwhile, mRNA level of suppressor of cytokine signaling 3 (Socs3), critical for cytokine secretion linked to neuroinflammation, was significantly decreased in isolated astrocytes. Moreover, we performed Barnes maze test to evaluate spatial learning and memory of castrated-App-KI mice and wild-type mice at 18 months of age. Aged wild-type mice exhibited the tendency of increase in the number of errors by castration, but this worsening effect by castration was slight in aged App-KI mice. These results suggested that testosterone may regulate pro-/anti-inflammatory molecules in glial cells and affect neuroinflammation and perhaps cognitive dysfunction. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-166
近接依存性標識法によるC9orf72リピートRNA分解促進因子の同定
Proximity labelling proteomics identified a molecule which promotes degradation of FTLD/ALS causing C9orf72 repeat RNA.
*魚住 亮太(1)、森 康治(1)、後藤 志帆(1)、宮本 哲愼(1)、近藤 志都子(1)、山下 智子(1)、河邉 有哉(2)、池田 学(1)
1. 大阪大学大学院医学系研究科、2. 医療法人社団 澄鈴会 箕面神経サナトリウム
*Ryota Uozumi(1), Kohji Mori(1), Shiho Gotoh(1), Tesshin Miyamoto(1), Shizuko Kondo(1), Tomoko Yamashita(1), Yuya Kawabe(2), Manabu Ikeda(1)
1. Grad Sch Med, Osaka University, 2. Minoh Neuropsychiatric Sanatorium
Keyword: C9orf72, FTLD/ALS, hnRNPA3, Proximity labelling proteomics
Intronic GGGGCC repeat expansion in C9orf72 is a most frequent genetic cause of familial frontotemporal lobar degeneration (FTLD)/amyotrophic lateral sclerosis (ALS). Neurodegeneration in C9orf72 FTLD/ALS is assumed to be caused by the toxicities of transcribed repeat RNA itself and/or its repeat-associated non-AUG (RAN) translation products, dipeptide repeat protein (DPR). hnRNPA3 is a heterogeneous nuclear ribonucleoprotein (hnRNP) identified as a GGGGCC repeat RNA binding factor. Although hnRNPA3 itself does not have apparent RNA degrading ability, increased hnRNPA3 expression led to a reduction of repeat RNA and DPR production, while reduction of hnRNPA3 induced accumulation of repeat RNA. Thus, hnRNPA3 promotes the degradation of GGGGCC repeat RNA. However, the exact mechanism by which hnRNPA3 promotes repeat RNA degradation is unclear. Liquid-liquid phase separation (LLPS) is mediated through weak and dynamic electrostatic and/or hydrophobic intramolecular interactions. Since most hnRNPs including hnRNPA3 are known to undergo LLPS, we hypothesized that loose binding of hnRNPA3 with factors potentially involved in RNA degradation may facilitate hnRNPA3-mediated RNA degradation. To mark and purify these loose binding factors of hnRNPA3, we introduced a state-of-art APEX2 technology which allows selective biotin labeling of proximate molecules in one minute. Biotinylated proteins in hnRNPA3-APEX2 expressing cells were purified with streptavidin pull-down for subsequent LC-MS/MS analysis, followed by Gene Ontology analysis. As a result, several RNA binding proteins were identified as candidate proteins. With secondary siRNA screening, we revealed one RNA binding protein which may facilitate hnRNPA3-mediated RNA degradation, because reduction of its expression induced remarkable GGGGCC repeat RNA accumulation, as quantified by RT-qPCR and in situ hybridization. This study reveals degradation pathway of the pathogenic repeat RNA in C9orf72 FTLD/ALS. Our results have implications for the future development of therapies for FTLD/ALS treatment. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-167
タウの生理的軸索局在決定における２段階の脱リン酸化
Two-step dephosphorylation in the physiological axonal localization of tau
*東 優人(1,2)、角田 聡子(1,2)、宮坂 知宏(1,2)
1. 同志社大学生命医科学研究科神経病理学研究室、2. 同志社大学神経変性疾患研究センター
*Higashi Yuto(1,2), Kakuda Satoko(1,2), Miyasaka Tomohiro(1,2)
1. Department of Neuropathology, Faculty of Life and Medical Sciences, Doshisha University, Kyoto, Japan, 2. Center for Research in Neurodegenerative Diseases, Doshisha University, Kyoto, Japan
Keyword: Alzheimer's disease, Tau, Microtubule, Phosphorylation
アルツハイマー病の病理学的特徴の一つである神経原線維変化は、高度にリン酸化されたタウが神経細胞の細胞体で凝集蓄積したものである。タウは軸索に局在する微小管結合タンパク質であり、その微小管結合および安定化能はリン酸化により負に制御されると考えられている。最近、タウ凝集モデルマウスの解析より、細胞体への異常局在がタウ凝集の起点となることが示された。タウ病変の形成には生理的な軸索局在機構および微小管結合の破綻が関与する可能性が高い。
脳成熟に伴い、細胞体で翻訳されたタウは軸索へ運ばれるとともに微小管上で機能するようになる。同時に高リン酸化状態であったタウは脱リン酸化されることが知られており、初代培養神経細胞を用いた解析から、タウの適切なリン酸化状態が軸索局在の決定に必須であるとされている。しかし、現在までに、翻訳されたタウがどのように軸索局在し、微小管上で機能するようになるのか、またそれらに対し、タウのリン酸化状態がどのような影響をおよぼすのか、その詳細なメカニズムは不明である。本研究では生後発達期のマウスを用いて、タウの産生から軸索へ局在化するまでのリン酸化状態と局在の変化について生化学的、組織学的に解析することを目的とした。
生後7日目 (P7) から13日目 (P13) の海馬について解析した結果、タウはP7 までは高度にリン酸化されており、P9 にかけて急激に脱リン酸化、さらに P11 から P13にかけて２段階目の脱リン酸化が起きていることを明らかにした。この期間におけるタウのリン酸化状態と局在の関係について解析した結果、AT8陽性のリン酸化タウは P7 時点では細胞体、樹状突起を含む細胞全体に分布し、P9 の時点ではその局在が保たれたまま脱リン酸化されることが確認された。P11〜P13にかけて次第にタウの軸索局在化が観察された。
これより、タウの軸索局在決定時には２段階の脱リン酸化ステップがあり、タウの Ser202 と Thr205 の脱リン酸化は軸索局在化に先行することを明らかとした。 タウの正確な軸索輸送には、微小管または未知の細胞内因子との相互作用が想定される。さらに我々はタウの軸索局在化メカニズムの詳細を明らかにするため、現在我々のグループで確立した微小管分画法及び超解像度顕微鏡を用いた解析を行っている。これらの結果を含めて、タウのリン酸化制御と生理的な軸索局在化および微小管との相互作用についての考察を試みる。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-168
脳内でβアミロイドーシスを誘発する可溶な高分子量Aβ分子種は，ヒトアルツハイマー病脳においてはCAAの目立つケースで豊富に存在する
Soluble high-molecular-weight Aβ species inducing cerebral β-amyloidosis are abundant in Alzheimer brains with prominent cerebral amyloid angiopathy.
*内上 寛一(1,2)、箱崎 眞結(1)、松原 知康(3)、村山 繁雄(3,4)、齊藤 祐子(3)、戸田 達史(2)、橋本 唯史(1,5)、岩坪 威(1)
1. 東京大学大学院医学系研究科神経病理学、2. 東京大学大学院医学系研究科神経内科学、3. 東京都健康長寿医療センター高齢者ブレインバンク、4. 大阪大学連合小児発達学研究科、5. 国立精神神経医療研究センター神経研究所疾病研究第四部
*Hirokazu Uchigami(1,2), Mayu Kashiwagi-Hakozaki(1), Tomoyasu Matsubara(3), Shigeo Murayama(3,4), Yuko Saito(3), Tatsushi Toda(2), Tadafumi Hashimoto(1,5), Takeshi Iwatsubo(1)
1. Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, 2. Department of Neurology, Graduate School of Medicine, The University of Tokyo, 3. Department of Neurology and Neuropathology (the Brain Bank for Aging Research), Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology, 4. Brain Bank for Neurodevelopmental, Neurology and Psychiatric Disorders, United Graduate School of Child Development, Osaka University, 5. Department of Degenerative Neurological Diseases, National Institute of Neuroscience, National Center of Neurology and Psychiatry
Keyword: Alzheimer’s disease, cerebral amyloid angiopathy, soluble high-molecular-weight Amyloid β, meninges
1. Objectives
Soluble amyloid β (Aβ) oligomers with a size of ~200-300 kDa extracted from autopsied brains of patients with Alzheimer’s disease (AD) have been shown to exhibit a seeding potency to induce cerebral β-amyloidosis. We examined the differences in the amyloid-inducing potencies between Aβ seeds recovered from AD brains with abundant or minimal levels of amyloid angiopathy, aiming at comparing those from cerebral blood vessels and brain parenchyma.
2. Methods
Six autopsied human AD brains with abundant cerebral amyloid angiopathy (CAA) and five AD brains without immunohistochemically detectable CAAs were examined. Tris-buffered saline (TBS)-soluble fractions from the brain or meningeal tissues were separated by size-exclusion chromatography (SEC) on Superdex 75 column, and the concentrations of Aβ in each SEC-separated fraction were measured by enzyme-linked immunosorbent assay. The TBS-soluble, ~200-300 kDa Aβ-positive fractions were stereotaxically injected into the unilateral hippocampus of 8-month-old amyloid precursor protein (APP) transgenic (tg) mice, and Aβ deposition in the hippocampus was assessed after four months to evaluate the seeding capacity.
3. Results
TBS-soluble Aβ species extracted from AD brains were eluted into two fractions at ~200-300 and ~10-20 kDa. The levels of Aβ in the ~200-300 kDa fractions from AD brains with abundant CAA were significantly higher than those in AD brains without CAA (p = 0.024). The concentration of Aβ in the ~200-300 kDa fraction from meninges was considerably higher than that from the brain parenchyma. Intrahippocampal injection of the TBS-soluble ~200-300 kDa fraction induced a characteristic laminar pattern of Aβ deposition in the brains of APP tg mice.
4. Conclusions
TBS-soluble Aβ in the ~200-300 kDa fractions inducing cerebral β-amyloidosis were more abundantly extracted from AD brains with prominent CAA compared to those without CAA, suggesting that the major source of the soluble, seed-competent Aβ in AD brains is cerebrovascular amyloid deposits, rather than those in brain parenchyma. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-169
経路統合能測定法の開発
Development of an assay for path integration performance
*小池 力(1)、山下 真央(1)、高市 雄太(2)、添田 義行(1)、高島 明彦(1)
1. 学習院大学大学院自然科学研究科、2. 東京大学大学院農学生命科学研究科
*Riki Koike(1), Mao Yamashita(1), Yuta Takaichi(2), Yoshiyuki Soeda(1), Akihiko Takashima(1)
1. Grad Sch Sci, Gakushuin Uni, Tokyo, Japan, 2. Grad Sch Agric and Sci, Univ of Tokyo, Tokyo, Japan
Keyword: Alzheimer's disease, path integration, entorhinal cortex
アルツハイマー病（AD）患者の脳内で観察されるタウ病理はBraakステージに沿って進展し、嗅内野における神経原線維変化がADで最も早期に出現する病理学的特徴であることが示唆されている。タウ病理は認知機能の低下とよく相関することが報告されており、嗅内野ステージで病理の進展を食い止めることは認知機能低下の抑制につながると考えられる。しかし、根本的な問題として、嗅内野ステージではほとんどの場合、神経心理学的検査で無症状を示すため、非侵襲的な発見・治療介入が難しい。そのため、嗅内野機能低下を反映する簡易なマーカーが必要である。そこで、嗅内野に存在するグリッド細胞に注目した。これは自身の進んだ距離や方向の情報を得るGPSの役割をしており、その活動は自己を定位する能力である経路統合に関連する。ADリスクの高い被験者で経路統合能の低下が観察されており、また、変異タウの過剰発現マウスで、グリッド細胞の周期性の崩壊が報告されている。しかし、嗅内野におけるタウ病理の経路統合への影響は直接的に示されておらず、これらの関連を明らかにすることを目的に本研究を行った。動物は、タウオパチーモデルマウスとして凝集能の高い変異タウを過剰発現するPS19を用いた。リン酸化タウを検出するAT8抗体を用いた免疫組織染色により、各月齢でのPS19のタウの蓄積部位を同定した。また、これらのマウスの経路統合能を定量化するため、2本の直線迷路を直角に組み合わせたL字迷路試験を検討した。明条件下でマウスをスタートさせると、L字型の迷路を通ってゴールに設置した暗箱へと逃避していく。このようにマウスをトレーニングした後、迷路を外してマウスを再度スタートさせると、2つの直線経路を統合し、L字ではなく直線的な経路をとってゴールへ向かっていく。その軌跡をカメラで記録し、スタートとゴールを結ぶ最短経路を0度とした時、マウスが進んだ経路との角度の誤差を経路統合スコアと定義した。さらに、空間認知機能のコントロールとして海馬依存的なバーンズ迷路を行った。免疫組織学的な解析から、ほとんど病理が観察されない4ヶ月齢のPS19と比較して、6-7ヶ月齢の嗅内野で有意にリン酸化タウ陽性細胞体の数が増加した一方、海馬では有意な増加は観察されなかった。この6-7ヶ月齢のPS19マウスでL字迷路試験を行うと、野生型マウスと比較して誤差角度が有意に増加した。一方で、バーンズ迷路については、野生型及びPS19マウス間で空間学習・記憶に有意な差はなかった。以上より、マウスモデルにおける嗅内野のタウ病理は空間学習・記憶に影響を与えない一方、経路統合障害を引き起こすことが示唆された。この結果から、経路統合能測定は、嗅内野の機能低下を検出することで嗅内野へのタウ蓄積を検出するアルツハイマー病の最初期臨床マーカーとなることが考えられた。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-170
ダウン症関連遺伝子のアミロイドβ蓄積に及ぼす影響の検討
Effects of Down syndrome genes on amyloid-β accumulation in mice
*石原 慶一(1)、河下 映里(1)、西村 周秦(2)、斉藤 美知子(3)、左合 治彦(4)、山川 和弘(5)、秋葉 聡(1)、高田 和幸(2)
1. 京都薬科大学 病態生化学、2. 京都薬科大学 統合薬科学、3. 京都薬科大学 バイオサイエンス研究セ、4. 成育医療研究セ 周産期・母性診療セ、5. 名古屋市立大学大学院医学研究科 神経発達症遺伝学分野
*Keiichi Ishihara(1), Eri Kawashita(1), Kaneyau Nishimura(2), Michiko Saito(3), Haruhiko Sago(4), Kazuhiro Yamakawa(5), Satoshi Akiba(1), Kazuyuki Takata(2)
1. Dept of Pathol Biochem, Kyoto Pharm Univ, Kyoto, Japan, 2. Divi of Integ Pharm Sci, Kyoto Pharm Univ, Kyoto, Japan, 3. Bio-sci Res Centr, Kyoto Pharm Univ, Kyoto, Japan, 4. Centr for Maternal-Fetal, Neonat & Reproduct Med, Natl Centr for Child Health & Development, Tokyo, Japan, 5. Depart of Neurodevelop Dis Genet, Nagoya City Univ Grad Sch Med Sci, Aichi, Japan
Keyword: Down syndrome, Alzheimer's disease
Individuals with Down syndrome (DS), which is caused by triplication of human chromosome 21 (Hsa21), develop an early-onset Alzheimer’s disease (AD) with extraordinarily high incidence. Since amyloid-β is a product of the amyloid precursor protein (APP) coded in Hsa21, increased amount of APP is thought to be the cause of this early-onset of AD. However, the highly incidence of early-onset AD in DS people could not be explained by merely 1.5-fold increase in APP expression, other Hsa21 genes, therefore, likely modulate the development of AD. To understand the genetic mechanisms underlying early-onset AD observed in DS, we tried to identify the DS gene(s) to modify the development of AD. In the present study, we generated mouse models of AD in DS by crossing DS mouse models with partial trisomies to AD mouse model expressing mutated APP and mutated presenilin 1. At 9 months of age, amyloid-β aggregates in the hippocampus and cerebral cortex were surprisingly decreased in DS-AD mouse model with approximately 70 trisomic Hsa21 genes. In contrast, no suppressive effects were observed in other DS-AD mouse model with a shorter extra copy of Hsa21 genes (approximately 30 genes), suggesting that the remaining 40 genes include a putative suppressor of amyloid-β aggregation. We also found that APP transgene-associated mortality was suppressed in both DS-AD mouse models. These results indicate that APP transgene-associated mortality is independent to the accumulation of amyloid-β aggregates. Here we found certain gene(s) in Hsa21 suppresses the amyloid-β accumulation and mutated APP-associated mortality. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-171
タウタンパク質のスプライシング制御を目的とするASOの開発と治療応用可能性について
Identification of antisense oligonucleotides that regulate splicing of tau proteins and their potential therapeutic applications
*遠藤 邦幸(1,2)、佐橋 健太郎(1)、河合 香里(1)、藤岡 祐介(1)、勝野 雅央(1)、祖父江 元(3)、石垣 診祐(1,4)
1. 名古屋大学大学院医学系研究科神経内科学、2. 松蔭病院、3. 愛知医科大学、4. 名古屋大学脳とこころの研究センター
*Kuniyuki Iwata-Endo(1,2), Kentaro Sahashi(1), Kaori Kawai(1), Yusuke Fujioka(1), Masahisa Katsuno(1), Gen Sobue(3), Shinsuke Ishigaki(1,4)
1. Dept Neurology, Nagoya Univ Grad Sch Med, Nagoya, Japan, 2. Matsukage Hospital, Nagoya, Japan, 3. Aichi Med Univ, Nagakute, Japan, 4. Brain Mind Res Cent, Nagoya University, Nagoya, Japan
Keyword: 4R-Tau associated tauopathies, frontotemporal lobar degeneration (FTLD), Fused in sarcoma (FUS), ENA-antisense oligonucleotides (ENA-ASO)
Background: Accumulations of 4R-tau predominant aggregation is particularly associated with frontotemporal lobar degeneration (FTLD), progressive supranuclear palsy (PSP), and cortico-basal degeneration (CBD). Previously, we showed that 4R-tau dominant status caused by loss of Fused in sarcoma (FUS) or Splicing factor, proline- and glutamine-rich (SFPQ) induced FTLD-like behaviors, reduced adult neurogenesis, accumulation of phosphorylated tau, and hippocampal atrophy in adult mice. Methods: We developed 2'-O, 4'-C-ethylene-bridged nucleic acid (ENA) modified antisense oligonucleotides (ASOs) which enabled to promote skipping of MAPT exon 10. We first screened for ENA-ASOs skipping exon 10 on MAPT by in vitro experiments. ASO was administered intracerebroventricularly as a single dose to humanized tau mice, and the optimal dose was determined by the maximum effect for exon 10 skipping. The course of inflammatory response induced by ASO administration was confirmed by immunostaining using GFAP and Iba1. Furthermore, we administered ASOs to FUS-silenced humanized tau mice (FUS KD mice) and confirmed its effects on behavior alterations. Results and Discussion: We selected NK-18 as the most potent ENA-ASO which reduced the 4R-tau/ 3R-tau ratio. Intracerebroventricular administration of NK-18 normalized the imbalanced 4R/3R-tau ratio in the brains of FUS KD mice. It also ameliorated the disease phenotypes including the aberrant behaviors and neurodegeneration in FUS KD mice. NK-18 exhibited its broad distribution throughout the brain and showed more potent and persistent ability for splicing correction which prolonged up to 2 years by the single intracerebroventricular administration of NK-18 than the 2'-O-methoxyethyl (MOE)-modified ASO (MOE-NK-18) which has the same sequences as NK-18. These findings support the therapeutic potential of ENA-ASO targeting MAPT exon 10 for 4R-tau associated tauopathies. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-172
APPswe-PS1ΔE9アルツハイマー病モデルマウスにおける海馬ニューロン新生の減少解明
Stage-specific reduction of hippocampal neurogenesis in APPswe-PS1ΔE9 Alzheimer's Disease model mice
*田村 理佐子(1)、林 大貴(1)、雷 晨旭(1)、李 昊煒(1)、久恒 辰博(1)
1. 東京大学大学院新領域創成科学研究科
*Risako Tamura(1), Daiki Hayashi(1), Chenxu Lei(1), Haowei Li(1), Tatsuhiro Hisatsune(1)
1. Graduate school of frontier sciences the university of Tokyo
Keyword: Alzheimer’s Disease, Neurogenesis
Alzheimer's disease(AD) is the most common form of dementia characterized by memory loss and cognitive impairment. The main causes are known to be the senile plaques formed by aggregation of amyloid-β(Aβ) and the neurofibrillary tangles formed by aggregation of hyperphosphorylated tau. Furthermore, recent studies have shown a decrease in new born neurons in AD patients with the progression of AD pathology (Moreno-Jiménez et al., 2019). Therefore, it is possible that there is a close relationship between AD pathology and the reduction of new born neurons. In hippocampus, the production and development of new born neurons occurs in the dentate gyrus, and after maturation, they are integrated into the hippocampal memory circuit, where they have specific functions and are thought to play an important role in learning and memory. A mouse model of AD that overexpresses Aβ peptides(APPswe-PS1ΔE9), has been reported to show a decrease in nestin-GFP expressing cells (Zeng et al., 2016), and a reduced maturation of new born neurons (Matsuda and Hisatsune, 2017). However, the mechanism of how Aβ peptides contributes to the reduction of hippocampal neurogenesis is still unknown. In order to study the mechanism of hippocampus neurogenesis impairment, we have also generated triple transgenic model mice in which neural transmission of new born neurons is specifically inhibited by the expression of tetanus toxin light chain (NBN-TeTX). We have found that the number of neural progenitor cells didn’t change, but there was impairment during the maturation of new born neurons in NBN-TeTX mice. Taken together, the decrease in the number of neural progenitor cells may be unique to AD model mice. To clarify the mechanism, we generated a new transgenic mice model (APP-NBN), which had been crossed between APPswe-PS1ΔE9 and NBN-TeTX mice. We found that APP-NBN mice showed a trend more impairment in reversal learning of spatial reference memory in Morris water maze test. We will further study the role of Aβ peptides in neurogenesis impairment in NBN-TeTX mice (APP-NBN) by using immunohistochemistry staining to examine the stage-specific alteration of cell proliferation, cell death, and neuronal maturation of new born neurons in AD mice. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-173
タウの蓄積およびタウオパチー病理形成が微小管安定性に与える影響
Impacts of Tau Accumulation and Tauopathy Pathogenesis on Microtubule Stability
*辰本 彩香(1,2)、宮坂 知宏(1,2)
1. 同志社大学大学院 生命医科学研究科 神経病理学研究室、2. 同志社大学神経変性疾患研究センター
*Ayaka Tatsumoto(1,2), Tomohiro Miyasaka(1,2)
1. Department of Neuropathology, Faculty of Life and Medical Sciences, Doshisha University, Kyoto, Japan, 2. Center for Research in Neurodegenerative Diseases, Doshisha University, Kyoto, Japan
Keyword: TAUOPATHY, ALZHEIMER'S DISEASES, MICROTUBULE STABILITY, MICROTUBULE ASSOCIATED PROTEINS
微小管結合タンパク質タウの凝集を伴う神経変性疾患を総称してタウオパチーという。タウ凝集体が生じる部位や頻度が症状と相関することから、タウ凝集メカニズムの解明はタウオパチー神経変性機構の理解とともに創薬の鍵になると考えられる。タウは健常脳においても豊富に存在しており、そのほとんどは微小管と結合している。一方、病理学的に凝集したタウは高度にリン酸化され、微小管への結合能が消失している。興味深い事にタウオパチー神経細胞内では微小管の消失が報告されている。そこで本研究ではタウの病理形成が微小管安定性に与える影響を明らかにするため、タウオパチーモデルマウス脳におけるタウの微小管結合および微小管安定性について解析した。
PS19マウスはP301S変異ヒトタウをプリオンプロモーター下で発現させたモデルで、6ヶ月齢ごろからタウ凝集体形成が認められる。3ヶ月齢（病理形成前）と11ヶ月齢（寿命のエンドポイント）の大脳皮質を用いて、当グループにより確立した微小管分画法により可溶性チューブリン画分 (FT)・動的微小管画分 (LM)・安定型微小管画分 (SM) に分画した。各画分に回収されたチューブリンと各種微小管結合タンパク質 (MAPs) についてWestern blotting法により定量した。
その結果、3ヶ月齢と11ヶ月齢の野生型およびPS19マウスにおいて、各画分中に回収されるチューブリンの割合（FT 9.9％、LM 55.5％、SM 34.6％）に差は認められなかった。すなわち、タウの蓄積および凝集体形成に関わらず、微小管の安定性は変化しないことが明らかとなった。3ヶ月齢のPS19マウス脳において、内在性マウスタウはFT 16.4％、LM 22.5％、SM 61.1％であるのに対し、外来性ヒトタウは FT 22.2％、LM 28.9％、SM 48.9％であった。これより、神経細胞内におけるタウの増加は微小管への結合性の低下をもたらすことを明らかとした。さらに11ヶ月齢について解析した結果、SM が減少し、FT に回収される遊離型タウの割合が有意に増加した。タウ以外のMAPs について解析した結果、MAP7はPS19マウス脳において SM の割合が減少し、LM が増加した。一方、タウ欠損マウスにおいては SM の割合が増加し、LM が減少したことから、MAP7は安定型微小管上でタウと競合する可能性が示された。
以上の結果より、PS19 マウス脳内において過剰発現したヒトタウは、加齢とともに微小管結合が低下し凝集体形成に至ることが示唆された。また、タウの過剰は微小管の安定性自体には影響を与えないが、一部他の MAPs の機能に影響を与える可能性が示された。微小管安定化機能を持つとされる MAPs の微小管結合様式変化が微小管安定性に影響しないことから、タウオパチー神経細胞における微小管の消失は、タウ凝集の下流ではなく独立した病理変化である可能性が考えられる。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-174
神経細胞の細胞老化に対する生理活性ペプチド因子ヒューマニンの影響
The effect of Humanin, a bioactive peptide, on cellular senescence in neurons
*小塚 彩里(1)、鈴木 弾(1)、新倉 貴子(1)
1. 上智大学
*Ayari Kozuka(1), Dan Suzuki(1), Takako Niikura(1)
1. Sophia University
Keyword: CELLULAR SENESENCE, PEPTIDE, HUMANIN
Cellular senescence is a permanent proliferation arrest which occurs in response to various endogenous and exogenous stresses. Cellular senescence contributes to tissue aging as well as progress of age-associated diseases such as Alzheimer’s diseases (AD). Senescent cells exhibit distinct phenotypes including the activation of DNA damage response, cell cycle arrest, activation of senescence-associated secretory phenotype, and apoptosis resistance. Some of these senescence markers were observed in AD brains. Humanin (HN) is a 24-residue polypeptide and initially identified as a neuroprotective factor against AD-related insults. HN suppresses neuronal death caused by amyloid beta, an AD-associated cytotoxic insult, in vitro and ameliorates memory deficit of AD mouse models. HN enhances mitochondrial ATP production in several types of cells including neurons. HN is a secretive peptide and the HN level in circulation decreases age-dependently in rodents and human. It is thus hypothesized that HN may have a role in preventing aging. In this study, we assessed the effect of S14G-HN, a highly potent HN derivative, on cellular senescence in neurons. Using neuronal differentiated PC12 cells, cell senescence was induced by glutamate and assessed by staining of senescence-associated β-galactosidase (SA-β-gal), a marker of senescence. S14G-HN dose-dependently suppressed the increase in the number of SA-β-gal positive cells. We obtained the similar result using primary neurons. Memantine, an antagonist of NMDA receptor, also reduced the number of SA-β-gal positive cells. When both S14G-HN and memantine were used together, an additional effect was observed. These results suggest the potential role of HN in suppressing aging-related cellular senescence. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-175
アルツハイマー病におけるミトコンドリア障害とタウ病理との関連
Mitochondrial complex I abnormalities is associated with tau pathology in Alzheimer’s disease
*寺田 達弘(1)、Therriault Joseph(2)、Kang Min Su(2)、松平 敬史(1)、武内 智康(1)、高嶋 浩嗣(1)、小尾 智一(3)、塚田 秀夫(4)、Rosa-Neto Pedro(2)、尾内 康臣(1)
1. 浜松医科大学、2. マギル大学、3. 静岡てんかん神経医療センター、4. 浜松ホトニクス中央研究所
*Tatsuhiro TERADA(1), Joseph Therriault(2), Min Su Kang(2), Takashi Matsudaira(1), Tomoyasu Bunai(1), Hirotsugu Takashima(1), Tomokazu Obi(3), Hideo Tsukada(4), Pedro Rosa-Neto(2), Yasuomi Ouchi(1)
1. Hamamatsu University School of Medicine, 2. The McGill University Research Centre for Studies in Aging, 3. Shizuoka Institute of Epilepsy and Neurological Disorders, 4. Hamamatsu Photonics KK
Keyword: Alzheimer's disease (AD), mitochondria, tau, PET
Introduction: Mitochondrial electron transport chain abnormalities have been reported in postmortem pathological specimens of Alzheimer’s disease (AD). However, it remains unclear how amyloid and tau are associated with mitochondrial dysfunction in vivo. The purpose of this study is to assess the local relationships between mitochondrial dysfunction and AD pathophysiology in mild AD using the novel mitochondrial complex I PET imaging agent [18F]BCPP-EF. Methods: Thirty-two amyloid and tau positive mild stage AD dementia patients (mean age ± SD: 71.1 ± 8.3 years) underwent a series of PET measurements with [18F]BCPP-EF mitochondrial function, [11C]PBB3 for tau deposition, and [11C]PiB for amyloid deposition. Age-matched normal control subjects were also recruited. Inter and intrasubject comparisons of levels of mitochondrial complex I activity, amyloid and tau deposition were performed. Results: The [18F]BCPP-EF uptake was significantly lower in the medial temporal area, highlighting the importance of the mitochondrial involvement in AD pathology. [11C]PBB3 uptake was greater in the temporo-parietal regions in AD. Region of interest analysis in the Braak stage I-II region showed significant negative correlation between [18F]BCPP-EF SUVR and [11C]PBB3 BPND (R=0.2679, p=0.04), positive correlations between the [18F]BCPP-EF SUVR and Wechsler Memory Scale-Revised logical memory score (R=0.2078, p=0.022), but not [11C]PiB SUVR. Conclusions: Our results indicated that mitochondrial complex I is closely associated with tau load and cognitive decline. The absence of association between mitochondrial complex I dysfunction with amyloid load suggests that mitochondrial dysfunction in the trans-entorhinal and entorhinal region is a reflection of neuronal injury occurring in the brain of mild AD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-176
アルツハイマー病および血管性認知症における海馬の変容
Pathological alterations of hippocampus in Alzheimer disease and vascular cognitive impairmet
*戸田 真太郎(1)、柳田 成史(1)、桒田 康宏(1)、馬場 麻悠子(1)、高山 直樹(1)、安田 謙(1)、眞木 崇州(1)、高橋 良輔(1)
1. 京都大学医学部付属病院医学研究科 臨床神経学
*Shintaro Toda(1), Seizi Yanagida(1), Yasuhiro Kuwata(1), Mayuko Baba(1), Naoki Takayama(1), Ken Yasuda(1), Takakuni Maki(1), Ryosuke Takahashi(1)
1. Department of Neurology, Graduate School of Medicine, Kyoto University Hospital
Keyword: Alzheimer, Hippocampus, Oligodendrocytes, Blood brain barrier
[Objective]The hippocampus is crucial for cognitive function and recent studies have shown the alterations of hippocampus at early stage of cognitive impairment. Blood brain barrier (BBB) disruption in hippocampus was shown to be detected in mild cognitive impairment (MCI) patients, suggesting that BBB breakdown is an early biomarker of human cognitive dysfunction. Additionally, neuronal hyperactivity in hippocampus was observed at early stage of Alzheimer's disease (AD) model mice. we previously reported that chronic cerebral hypoperfusion, which is frequently comorbid with AD, resulted in hippocampal degeneration with BBB disruption in mice. Neurodegeneration and synaptic loss with Aβ and tau deposition is a central feature of AD. However, neuron-centric view cannot explain all aspects of AD. Among a variety of cells in the brain and outside the brain, oligodendrocyte lineage cells (OLs) are one of pivotal cell types which affect the AD pathogenesis. Besides well-known role of reservoir for oligodendrocytes (OLGs) which support neurons, oligodendrocyte precursor cells (OPCs) regulate neurovascular function, including blood BBB integrity, in diverse ways. Recent single-nucleus RNA-seq analysis demonstrated the robust alterations of gene expressions in OLs in AD patients. Vascular cognitive impairment (VCI) is a broad concept that covers the full spectrum from vascular mild cognitive impairment (vascular MCI) to vascular dementia and also includes cases with mixed vascular and AD pathologies. There are several proof-of-concept studies which examined the hypotheses that dysfunctional OLs and demyelination may contribute to the onset and progression of VCI. However, most of these hypotheses still remain unproven.Therefore, the aim of this study is to examine the early alterations of hippocampus in relation to pathological OLs in AD and VCI model mice. [Methods]We examined the pathology of hippocampus in AD model mice with and without chronic cerebral hypoperfusion, which is produced by bilateral common carotid artery stenosis (BCAS). Moreover, we evaluated the effect of blood–derived molecules such as fibrinogen and thronbin on OLs for in vitro study. [Results]BCAS1, NG2, and Olig2-OLs were located in close proximity with Aβ accumulation. Aberrant neuronal activity and demyelination with vascular abnormalities and BBB disruption in hippocampus was observed in BCAS-operated AD model mice. [Conclusions]The present study suggests that early pathological alterations in hippocampus with compromised OLs would be therapeutic target for VCI. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-177
Aβ病理モデルマウスを用いたアルツハイマー病血液バイオマーカーリン酸化タウ181，217，231の脳内局在解析
Brain pathologies associated with phospho-tau 181, 217 and 231, fluid biomarkers for Alzheimer’s disease, in App knock-in mouse models of Aβ amyloidosis
*廣田 湧(1,2)、榊原 泰史(1)、茨木 京子(1)、竹井 喜美(1)、飯島 浩一(1,3)、関谷 倫子(1,3)
1. 国立長寿医療研究センター 神経遺伝学研究部、2. 日本学術振興会特別研究員PD、3. 名古屋市立大学大学院薬学系研究科
*Yu Hirota(1,2), Yasufumi Sakakibara(1), Kyoko Ibaraki(1), Kimi Takei(1), Koichi M Iijima(1,3), Michiko Sekiya(1,3)
1. Dept. of Neurogenetics, National Center for Geriatrics and Gerontology, 2. JSPS Research Fellow (PD), 3. Grad Sch Pharmaceutical Sciences, Nagoya City University
Keyword: Alzheimer’s disease, Amyloid-β, Biomarker, Phospho-tau
Tau proteins phosphorylated at T217, T231 as well as T181 in plasma and cerebrospinal fluid (CSF) are promising diagnostic biomarker for Alzheimer’s disease (AD). Changes of these phospho-tau (p-tau) levels in fluid precede neurofibrillary tangle (NFT) pathology and are correlated with amyloid-β (Aβ) burden in human brains. More recent reports demonstrate that p-tau 217 is more sensitive than p-tau 181. However, brain pathology reflecting these p-tau species remains elusive. Here, we examined intracerebral localization of biomarker p-tau in App knock-in mice with Aβ plaques but no NFT pathology (AppNLGF), increased Aβ levels but no Aβ plaque (AppNL) and wild-type mice. Immunohistochemical analysis revealed that signals of p-tau 217 and p-tau 231 were only detected in the brains of AppNLGF mice. These p-tau species appeared as puncta signals around Aβ plaques, overlapped with tau pathology marker phospho-tau S202/T205/S208 (AT8 epitope), colocalized with postsynaptic marker PSD95 but not with presynaptic or glial markers, and increased upon aging. In contrast, p-tau 181 signals were detected in the axonal structures in AppNLGF, AppNL and wild-type mouse brains, while in AppNLGF mice, p-tau 181-positive aberrant cluster-like structures were detected around Aβ plaques, which were partially overlapped with p-tau 217. Signals of neurofilament light chain (NfL), a biomarker for neurodegeneration, were similar between AppNLGF, AppNL and wild-type mice. These results suggest that p-tau 217, 231 and a part of 181 reflect synaptic pathology, while p-tau 181 also represents aberrant axonal structures induced by Aβ plaques. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-178
Identifying cellular and proteomic phenotypes across human Alzheimer’s disease hippocampal tissue by multiplexed ion beam imaging
*Cannon Bryan(1)、Vijayragavan Kausalia(1)、Tebaykin Dmitry(1)、Mrdjen Dunja(1)、Oliveria JP(1)、Greenwald Noah(1)、Baranski Alex(1)、Angelo Mike(1)、Montine Tom(1)、Bendall Sean(1)
*Bryan Cannon(1), Kausalia Vijayragavan(1), Dmitry Tebaykin(1), Dunja Mrdjen(1), JP Oliveria(1), Noah Greenwald(1), Alex Baranski(1), Mike Angelo(1), Tom Montine(1), Sean Bendall(1)
1. Stanford University
Keyword: Alzheimer's Disease, Imaging, Proteomics, Microglia
Identifying cell-based and regional phenotypes in human brain tissue is a necessary task to understand neurodegenerative pathologies in Alzheimer’s disease. Using MIBI (multiplexed ion-beam imaging) we have simultaneously quantified tissue-level expression of 40 different targets encoding a deep set of neurological phenotypes with nanometer scale spatial resolution. To localize protein expression to specific cells or tissue restricted regions, we use a combination of single cell and single object segmentation methods. Applying this technique to multiple MIBI scans of archival human hippocampus cross-sections, we are able to identify the spatial distribution and expression heterogeneity of glial, neuronal, and endothelial cell types and proteopathies in both cognitively healthy and diseased human brain. Using off-the-shelf and in-house developed computational strategies we are able to isolate unique protein-disease trends in traditional hippocampal neuroanatomy, particularly identifying tau-tangle targeting by microglia in severe Alzheimer's within the CA1 region. Additional work using cross-sample clustering algorithms derived common areas of disease and healthy tissue independent of AD severity. Identification of neurons able to survive in these proteopathy-associated regions may lead to additional work testing the functional significance of the descriptive work outlined here. We believe this will be a straight forward approach to discretize and select features for multiplexed imaging technologies as applied to human brain tissue in both degenerative and non-degenerative contexts. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-179
ストレス下におけるアルツハイマーモデルマウスを用いた脳血流量測定法の探索
Search for methods to measure cerebral blood flow in Alzheimer's disease model mouse under stress
*井上 由理子(1)、江連 博光(1)、伊藤 純治(2)、澤 智華(3)、高柳 雅朗(4)、高木 孝士(5)、馬目 佳信(6)、井上 明男(7)、大塚 成人(1)
1. 昭和大学 医学部 解剖学講座 肉眼解剖学部門 、2. 昭和大学 保健医療学部 理学療法学科 、3. 昭和大学 医学部 解剖学講座 顕微解剖学部門 、4. 埼玉県立大学 保健医療福祉学部 共通教育科、5. 昭和大学 電子顕微鏡室、6. 慈恵会医科大学 基盤研究施設、7. 京都大学 医学研究科 附属脳機能総合研究センター
*Yuriko Inoue(1), Hiromitsu Ezure(1), Jyunji Ito(2), Chika Sawa(3), Masaaki Takayanagi(4), Takashi Takaki (5), Yoshinobu Manome(6), Akio Inoue(7), Naruhito Otsuka(1)
1. Dep Anat, Showa Univ Sch Med, Tokyo, Japan, 2. / School of Nurs. Reh. Sci, Showa Uni Nursing, Kanagawa, Japan , 3. Dep Anat, Showa Univ Sch Med. Tokyo, Japan, 4. Saitama Prefectural University, Saitama, Japan , 5. Div. Electron Microscopy Showa Univ, Tokyo, Japan , 6. Core Research Facilities for Basic Science, Research Center for Medical Sciences, Jikei Uni, Tokyo, Japan, 7. Human Brain Res. Center, Kyoto Uni. Med, Kyoto, Japan
Keyword: Alzheimer’s disease model mouse, Mn-MRI
【Background】Mn ions flow into neurons with Ca ions upon nerve stimulation, and Mn ions enhance the T1 signal of MRI, so it is expected to be possible to see neural excitation by MRI. However, while measurements have been made at the cellular level, actual Mn levels have not been measured using MRI in mice.
【Purpose】There is a point at which Mn ions leak out of the cell with continued nerve stimulation; intraperitoneal injection of Mn was found to increase concentrations in the brain for approximately 3 hours. Mn-MRI measurements were performed to investigate the site of stimulation in the brain of Alzheimer's model mice as well as in neurons.【Results】We studied the cause of Alzheimer's disease using transgenic mouse expressing a large amount of amyloid.
The transgenic mouse shows the decrease of memory by the Y-maze test. However, no clear anatomical difference was observed between transgenic and control mice.
As nerve cells uptake Mn ions depending on nerve activity, we injected Mn ions into the abdominal cavity of mouse and examined Mn in the brain using Bruker 9.4T MRI machine with cryoprobe. Magnetic resonance imaging (MRI), uses a strong magnetic field and radio waves to generate images of parts of the body. The contrast of image is produced by the difference of stability of activated state produced by radio wave. Nerve cells uptake Ca ions during neuronal excitement. Ca ions were monitored using Ca indicator, Fluo4 which binds with Ca ions and the complex is fluorescent. Fluo4 is soluble and does not enter cells. Then, Fluo4-AM which being lipid soluble and pass through the cell membrane was used. Fluo4-AM is hydrolyzed into Fluo4 in the cytoplasm and stay in the cell. Mn ions also pass through the Ca channel and enter nerve cells. Mn ions bind more strongly with Fluo4, but the Mn-Fluo4 complex is not fluorescent. Hippocampal neurons were isolated from Wistar rats at 18 days of gestation (E18). The cells after DIV (Days in Vitro) 21 are used for the experiments. Activated state of proton in free water is rather stable compared to that in lipid and proteins. As Mn ions destabilize the activated state, T1 intensity is activated by Mn ions. The transgenic mouse shows the decrease in Mn incorporation to the dentate gyrus of hippocampus and the cerebrum including cingulate cortex, motor and sensory area.
【Discussion】
Stimulation site measurements were possible from Mn-MRI in the brains of Alzheimer's model mice. However, it was also suggested that quantification is needed. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-180
NSF欠損マウスは自閉症様な異常を示す
N-ethylmaleimide-sensitive factor (NSF) deficient mice exhibit autism like phenotypes
*謝 敏かく(1,2,3)、岩田 圭子(1,2,3)、石川 保幸(4)、村田 航志(5)、深澤 有吾(1,2,5)、松﨑 秀夫
1. 福井大学 子どものこころの発達研究センター 脳機能発達研究部門、2. 福井大学 ライフサイエンスイノベーションセンター、3. 大阪大学大学院 大阪大学・金沢大学・浜松医科大学・千葉大学・福井大学 連合小児発達学研究科、4. 前橋工科大・システム生体工学、5. 福井大学大学院 医学系研究科 脳形態機能学領域
*Min Jue Xie(1,2,3), Keiko Iwata(1,2,3), Yasuyuki Ishikawa(4), Koshi Murata(5), Yugo Fukazawa(1,2,5), Hideo Matsuzaki
1. Div Dev Mental Func, Res Center Child Mental Dev, Univ of Fukui, 2. Life Sci lnnovation Center, Univ of Fukui, 3. United Grad Sch Child Dev, Osaka Univ, Kanazawa Univ, Hamamatsu Univ Sch Med, Chiba Univ, Univ of Fukui, 4. Dept Sys Life Engineering, Maebashi Inst Tech, Gunma, Japan, 5. Div Brain Struc Func, Dept Morphol Physiol Sci, Grad Sch Med Sci, Univ of Fukui, Fukui, Japan
Keyword: Autism, NSF, SERT, AMPAR
Autism spectrum disorder (ASD), characterized by profound impairment in social interactions and communication skills, is the most common neurodevelopmental disorder. Many studies on the mechanisms underlying the development of ASD have focused on the serotonergic system; however, these studies have failed to completely elucidate the mechanisms. We previously identified N-ethylmaleimide-sensitive factor (NSF) as a new serotonin transporter (SERT)-binding protein and described its importance in SERT membrane trafficking and uptake in vitro. In the present study, we generated Nsf+/- mice and investigated their behavioral, neurotransmitter, and neurophysiological phenotypes in vivo. Nsf+/- mice exhibited abnormalities in sociability, communication, repetitiveness, and anxiety. Additionally, Nsf loss led to a decrease in membrane SERT expression in the raphe and accumulation of glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors at the synaptic membrane surface in the hippocampal CA1 region. We found that postsynaptic density and long-term depression were impaired in the hippocampal CA1 region of Nsf+/- mice. Taken together, these findings demonstrate that NSF plays a role in synaptic plasticity and glutamatergic and serotonergic systems, suggesting a possible mechanism by which the gene is linked to the pathophysiology of autistic behaviors. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-181
レット症候群ならびにMeCP2重複症候群患者由来iPS細胞株からの大脳皮質オルガノイドの作製
Generation of cortical organoids derived from Rett syndrome and MeCP2 duplication syndrome iPS cell lines
*白石 敦士(1)、濱村 祐輝(1)、井上 貴仁(1)、小野 恵秀(1)、有岡 祐子(2)、奥村 啓樹(2)、二村 隆史(1)、尾崎 紀夫(2)
1. 大塚製薬（株）中枢神経疾患研究所、2. 名古屋大学大学院医学系研究科精神医学分野
*Atsushi Shiraishi(1), Yuki Hamamura(1), Takahito Inoue(1), Toshihide Ono(1), Yuko Arioka(2), Hiroki Okumura(2), Takashi Futamura(1), Norio Ozaki(2)
1. Department of CNS Research, Otsuka Pharmaceutical Co., Ltd., Tokushima, Japan, 2. Department of Psychiatry, Nagoya University Graduate School of Medicine, Nagoya, Japan
Keyword: Autism, MeCP2, Organoids, Induced pluripotent stem cells (iPSCs)
Some psychiatric disorders, including autism spectrum disorder (ASD), are highly heritable, and several genetic risk factors of these disorders have been reported. Although genetic-engineered animal disease models induce behavioral phenotypes similar to a human condition, these models alone are still poor predictors of human biology, especially in central nervous system (CNS) diseases in clinical trials.
Human induced pluripotent stem cells (iPSCs) contain donor genetic background and have the potential to generate any type of cell or tissue except extraembryonic tissues. Therefore, the iPSC-derived neurons and tissues, what we call brain organoids, are expected to be useful for CNS disease modeling and drug discovery. We can generate region-specific brain organoids from human iPSCs. The organoids can recapitulate developmental processes, the organization consisting of multiple cells and some specific function of the in vivo organs.
The neurodevelopmental disorder Rett syndrome (RTT) and MeCP2 duplication syndrome (MDS) are caused by mutation and duplication of the X-linked methyl CpG binding protein-2 (MeCP2) gene, respectively. RTT and MDS also cause psychiatric symptoms such as ASD-like poor communication skills, repetitive behaviors and severe mental retardation. MeCP2 plays a multifaceted role in gene expression regulation (transcriptional repression/activation) and chromatin organization. Therefore, MeCP2 is required for proper brain development and some RTT patients acquire microcephaly.
Here, we report that we can generate cortical organoids derived from healthy control (HC), RTT (MeCP2 R255X), RTT isogenic control and MDS iPSCs. The size of RTT organoids was far smaller than the other cell lines. HC, RTT isogenic control and MDS organoids represented spatial organization of cortical neuron subtypes. We also found that the length of neurites of MDS dissociated neurons was shorter than that of neurites of HC dissociated neurons.
Using the organoids and neurons derived from human patient-derived iPSCs, we are going to establish disease models and a platform for drug discovery for psychiatric disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-182
胎児期に着目した自閉スペクトラム症研究
Autism spectrum disorder research focusing on the fetal period
*井田 瑛理香(1)、笠原 好之(1)、吉田 千尋(1)、伊藤 崚馬(1)、齋藤 昌利(1)、木村 芳孝(1)
1. 東北大学大学院医学系研究科
*Erika Ida(1), Yoshiyuki Kasahara(1), Chihiro Yoshida(1), Ryoma Ito(1), Masatoshi Saito(1), Yoshitaka Kimura(1)
1. Grad Sch Med, Univ of Tohoku, Sendai, Japan
Keyword: Autism, Fetus, Autonomic nervous system, Electrocardiogram
Autism spectrum disorder (ASD) is a developmental disorder that causes difficulty in interpersonal relationships and qualitative impairments in communication. Although the etiology of ASD is still unknown, one of the environmental factors is maternal immune activation (MIA) due to inflammation during pregnancy; MIA is mainly caused by inflammation due to infection and, in severe cases, is known to increase the risk of developing ASD in children. It has been observed that pups born from MIA model mice exhibit ASD-like behaviors after reaching adulthood.On the other hand, it has been reported that the autonomic nervous system function, as assessed by electrocardiography, is changed in children with ASD. Also, since early education and environmental adjustment can alleviate the symptoms of ASD, diagnosis in utero may help reduce the burden on ASD patients. Therefore, in this study, we compared the autonomic nervous system activity of fetuses in MIA-infected mother mice by using bacterial and viral infection models.Fetuses in which the mother mouse received saline were used as controls. MIA-induced ASD model fetal mice were created by administering polyinosinic-polycytidylic acid (Poly(I:C), viral infection model) and Lipopolysaccharide (LPS, bacterial infection model), respectively, to the mother mice.The mating day was defined as Embryonic day(E)0. MIA was induced at E12.5, and at E18.5, mother mice were laparotomized under anesthesia, and needle electrodes were placed on the fetus for electrocardiographic measurements. Power spectrum analysis was used to evaluate the function of the fetal autonomic nervous system.Compared to the control group, high frequency (HF) power was reduced in both models, and low frequency (LF) power and short-term variability (STV) were significantly reduced in the bacterial infection model. This suggests that parasympathetic nervous activity is decreased in the viral infection model, and both parasympathetic and sympathetic nervous activities are decreased in the bacterial infection model. The increase in HR only in the viral infection model could be attributed to this difference in autonomic balance. To investigate the reason for the above difference, we will conduct comprehensive genetic analysis of the fetuses in the future. In addition, we will conduct behavioral experiments in pups born from MIA model mice to elucidate the pathogenesis of ASD and to investigate therapeutic methods. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-183
MEAを用いた自閉スペクトラム症患者由来iPS神経の神経活動評価
Evaluation of Neural activity of iPSC-derived neurons from Autism Spectrum Disorder donor with Micro electrode array.
*仲山 智明(1)、瀬尾 学(2)、荒谷 知行(2)、東 基記(1)
1. 株式会社リコー、2. Elixirgen Scientific, Inc.
*Tomoaki Nakayama(1), Manabu Seo(2), Tomoyuki Aratani(2), Azuma Motoki(1)
1. RICOH COMPANY, LTD. , Kanagawa, Japan, 2. Elixirgen Scientific, Inc., Baltimore, US
Keyword: Autism Spectrum Disorder, Micro Electrode Array, Drug discovery, iPSC-derived neuron
【Background & Purpose】 The significant difference between animal models and humans is one of major reasons for project closures of drug discovery research and development in central nervous system (CNS) field. And, it is also well known that in vitro/ in vivo animal models are often insufficient to predict drug efficacies and toxicities in human. To overcome this challenge, many assay methods with the human induced pluripotent stem cell (iPSC) derived neurons are being reported. Multielectrode array (MEA) is one of the well-known methods to evaluate nerve functions. We have constructed the stable MEA evaluation system by co-culturing Quick-Neuron ™, which differentiated iPS cells into neurons by Quick-Tissue™ Technology, and primary human astrocytes. In order to confirm that the constructed MEA evaluation system can be applied to drug discovery screening in CNS field, we investigated the difference between healthy donor derived neurons and disease donor derived neurons. We used iPS cells from an Autism Spectrum Disorder (ASD) donor which is one of the major diseases of the central nervous system and compared the neuronal firing characteristics between from ASD patients and Healthy donor. 【Procedure】 Quick-Neuron™ and primary human astrocytes were co-cultured on Cytoview-MEA48 plates. Neuronal firing characteristics were evaluated over time using Maestro Pro for 6 weeks. 【Results】 From the 3rd week of culture, network bursts were detected in both from ASD patient and Healthy donor. At the 6th week, there was a specific difference in the shape of the raster plot, and a significant difference in the number and the duration of network burst was detected. 【Future work】 In the future, we will investigate the cause of this abnormal neuronal firing state identified in this study. 【Conclusion】 From this result, it was showed that this MEA evaluation system using Quick-Neuron ™ detect the specific differences in neuronal firing characteristics derived from ASD patients and healthy doner. It was suggested that this evaluation system would be a useful tool for drug discovery research with human iPSC derived neurons. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-184
自閉症者の白色ノイズへの交感神経応答は馴化する
People with autism spectrum disorder show habituation of the sympathetic response to whitenoise
*大石 悠貴(1)、糸井 千尋(2)、加藤 進昌(3)、柏野 牧夫(1)
1. NTTコミュニケーション科学基礎研究所、2. 国立障害者リハビリテーションセンター研究所、3. 昭和大学発達障害医療研究所
*Yuuki Ooishi(1), Chihiro Itoi(2), Nobumasa Kato(3), Makio Kashino(1)
1. NTT Communication Science Laboratories, 2. Research Institute of National Rehabilitation Center for Persons with Disabilities, 3. Medical Institute of Developmental Disabilities Research, Showa University
Keyword: autism spectrum disorder, sympathetic response, audition
It is well known that many people with autism spectrum disorder (ASD) have hyperacusis. To examine the effects of sound on humans, sympathetic responses are frequently measured. Our hypothesis is that individuals with ASD would show a nonhabituating sympathetic response to sound stimuli due to the symptom of hyperacusis, which no study has considered yet. In this study, we examined the difference in the degree of habituation of the sympathetic response to sound between people with ASD and people with typical development (TD). We recruited ten people with ASDs and eleven TDs. The transducer of a photoplethysmogram was attached to their second finger to measure the blood volume pulse response (BVPR), which is a kind of noradrenergic sympathetic response. Whitenoise was represented as a sound stimulus for 180 s. We divided this 180s-time region into four parts defined as follows: an orienting phase (t = 0–30 s), habituation phase 1 (t = 30–80 s), habituation phase 2 (t = 80–130 s), and habituation phase 3 (t = 130–180s). We found that the size of the BVPR in the orienting phase was not significantly different between the participants with ASD and TDs. The BVPRs of ASD participants were significantly smaller than those of the TDs in the habituation phases regardless of the phase. These results suggest that the sympathetic orienting response to whitenoise in ASD individuals is not different from that of TDs, whereas the degree of habituation is larger in the former, which is completely contrary to our hypothesis. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-185
ATF5はマウス大脳皮質におけるインターニューロンの維持に関与している。
ATF5 is involved in the maintenance of interneurons in the mouse cerebral cortex.
*中島 理子(1)、梅村 真理子(1)、喜屋武 諒(1)、高橋 滋(1)、高橋 勇二(1)
1. 東京薬科大学
*Riko Nakajima(1), Mariko Umemura(1), Ryo Kiyatake(1), Shigeru Takahashi(1), Yuji Takahashi(1)
1. Tokyo University of Pharmacy and Life Sciences
Keyword: interneuron, ATF5, cerebral cortex
Interneurons (INs) are inhibitory neurons that inhibit the activity of other neurons by releasing a neurotransmitter GABA from their axon terminals. In the cerebral cortex of mammalian brains, the bulk of the neuronal populations are composed of glutamatergic excitatory neurons and the remaining (~20–30%) are composed of GABAergic inhibitory interneurons. INs can be divided into several subtypes according to the proteins they express. In mouse models and patients with psychiatric disorders, IN dysfunction, disturbance of the IN subtype population, and decreased density have been reported, suggest that abnormalities in INs may be one of the causes of psychiatric disorders. Activating transcription factor 5 (ATF5) is a member of the cAMP response element binding protein (CREB)/ATF family of basic leucine zipper transcription factors. It is a stress responsive transcription factor under conditions such as ER stress and oxidative stress. It has been found that ATF5 is expressed in neurons in the cerebral cortex, striatum, hippocampus, and cerebellum in the adult mouse brain. ATF5 is essential for the proliferation and differentiation of progenitor cells and plays a neuroprotective role against ER stress. ATF5 knockout (ATF5-/-) mice exhibited behavioral abnormalities including hyperactivity in novel environments, increased anxiety-like behavior and reduced social interaction. We showed that the olfactory bulbs of ATF5-/- mice were smaller than that of wild type littermates at both neonatal and adult stages, and had an irregular laminar structure, especially in the glomerular layer (GL). In addition, a reduced number of interneurons has been observed in the GL of ATF5-/- mice. Next, immunostaining for GAD67, a GABA synthase, and subtypes such as parvalbumin, calbindin, and somatostatin was performed to compare the density of INs in the cerebral cortex. Compared to the ATF5+/+ cortex, the ATF5-/- cortex exhibited an altered distribution of INs in adults. Also, western blotting analysis showed that the expression level of the INs marker protein was decreased in lysates of ATF5-/- mouse cortices. These results suggest that ATF5 deficiency may affect the survival and maintenance of INs, indicating that ATF5 plays an important role in INs function. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-186
大脳皮質発達における自閉スペクトラム症関連遺伝子の役割
Role of autism spectrum disorder-associated gene in neocortical development
*臼井 紀好(1,2,3,4)、仲間 菜々子(1)、宮内 波奈(1)、土井 美幸(1)、入江 浩一郎(1)、小山 佳久(1,4)、中村 雪子(1,4)、松﨑 秀夫(2,5)、島田 昌一(1,2,3,4)
1. 大阪大学大学院医学系研究科神経細胞生物学講座、2. 大阪大学大学院連合小児発達学研究科、3. 大阪大学国際医工情報センター、4. 大阪精神医療センターこころの科学リサーチセンター依存症ユニット、5. 福井大学子どものこころの発達研究センター
*Noriyoshi Usui(1,2,3,4), Nanako Nakama(1), Hana Miyauchi(1), Miyuki Doi(1), Koichiro Irie(1), Yoshihisa Koyama(1,4), Yukiko Nakamura(1,4), Hideo Matsuzaki(2,5), Shoichi Shimada(1,2,3,4)
1. Dept Neurosci Cell Biol, Grad Sch Med, Osaka Univ, Osaka, Japan, 2. United Grad Sch Child Dev, Osaka Univ, Osaka, Japan, 3. Global Cent Med Eng Info, Osaka Univ, Osaka, Japan, 4. Addiction Res Unit, Osaka Psych Res Center, Osaka Psych Med Cent, Osaka, Japan, 5. Res Cent Child Ment Dev, Univ Fukui
Keyword: AUTISM SPECTRUM DISORDER, SOCIAL BEHAVIOR, MYELINATION, TRANSCRIPTOME
Zinc finger and BTB domain containing 16 (ZBTB16) plays the roles in the neural progenitor cell proliferation and neuronal differentiation during development, however how the function of ZBTB16 is involved in brain function and behaviors unknown. Here we show the deletion of Zbtb16 in mice leads to social impairment, repetitive behaviors, risk-taking behaviors, and cognitive impairment. To elucidate the mechanism underlying the behavioral phenotypes, we conducted histological analyses and observed impairments in thinning of neocortical layer 6 (L6) and a reduction of TBR1+ neurons in Zbtb16 KO mice. Furthermore, we found increased dendritic spines and microglia as well as developmental defects in oligodendrocytes and neocortical myelination in the prefrontal cortex (PFC) of Zbtb16 KO mice. Using genomics approaches, we identified the Zbtb16 transcriptome that includes genes involved in neocortical maturation such as neurogenesis and myelination, and both autism spectrum disorder (ASD) and schizophrenia (SCZ) pathobiology. Co-expression networks further identified Zbtb16-correlated modules that are unique to ASD or SCZ respectively. Our study provides insight into the novel roles of ZBTB16 in behaviors and neocortical development related to the disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-187
生後発達期大脳皮質のIL-17A受容体発現と母体免疫活性化ASDモデルにおけるその変化
IL-17RA expression in the cerebral cortex during postnatal development and its alternation in the ASD model of maternal immune activation
*佐々木 哲也(1,2)、Bao Peiyi(2)、大塚 優江(1)、洲崎 浩子(1)、岩田 卓(1,2)、武井 陽介(1,2)
1. 筑波大 医 生命医科学・解剖学・神経科学、2. 筑波大 院人間総合 ニューロサイエンス学位プロ
*Tetsuya Sasaki(1,2), Peiyi Bao(2), Masae Ohtsuka(1), Hiroko Suzaki(1), Suguru Iwata(1,2), Yosuke Takei(1,2)
1. Dept Anat & Neurosci, Fac Med, Univ of Tsukuba, Ibaraki, Japan, 2. Doctorate Program Neurosci, Grad Sch Comp Human Sci, Univ of Tsukuba, Ibaraki, Japan
Keyword: Autism Spectrum Disorder, Interleukin-17A, IL-17RA, Cerebral Cortex
Clinical studies have suggested that immune responses by helper T cells 17 (Th17 cells) are involved in the pathogenesis of autism spectrum disorders (ASD), schizophrenia, and depression. However, it remains unclear how the immune response induces organic changes in the nervous system, and the contribution of Th17 cells is not well understood. Interleukin (IL-)17A is a pro-inflammatory cytokine that binds to a receptor composed of a heterocomplex of IL-17RA and IL-17RC and activates downstream pathways. Previous studies have reported that Il17ra is mainly expressed in the cortical plate of mouse embryonic brain at 14.5 days of embryonic period and that IL-17A increased in the placenta by maternal poly(I:C) administration upregulates Il17ra mRNA expression. The expression of Il17a and its receptors in the adult central nervous system has been reported by several research groups but continues to be controversial. In this study, we investigated the expression of Il17ra mRNA in the primary somatosensory cortex during postnatal development from neonatal to young adult by in situ hybridization. Il17ra mRNA was strongly expressed in layer VI (both VIa and Vb) throughout the period examined. The expression was maximal at 14 days of age and decreased through adulthood. Double staining with cellular markers showed that it was expressed in about 60% of layer VI neurons. In addition, a decrease in Il17ra mRNA expression was observed at 14 days of age after maternal poly(I:C) administration. This study may provide the first clue to the function of IL-17A and IL17RA in central nervous system during the postnatal period. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-188
マウス胎生期のバルプロ酸曝露は脊髄でのミクログリアの増殖とアロディニアを引き起こす
Prenatal exposure to valproic acid induces proliferation of microglia in the spinal cord and mechanical allodynia in mice
*吾郷 由希夫(1,2)、今戸 瑛二(1,3)、Samnang Sun(2)、中村 庸輝(4)、中島 一恵(4)、森岡 徳光(4)、田熊 一敞(5)、木口 倫一(6)、浅野 智志(1,2)
1. 広島大学大学院医系科学研究科細胞分子薬理学、2. 広島大学歯学部、3. 広島大学大学院医系科学研究科歯科麻酔学、4. 広島大学大学院医系科学研究科薬効解析科学、5. 大阪大学大学院歯学研究科薬理学、6. 和歌山県立医科大学薬学部生体機能解析学
*Yukio Ago(1,2), Eiji Imado(1,3), Sun Samnang(2), Yoki Nakamura(4), Kazue Nakashima(4), Norimitsu Morioka(4), Kazuhiro Takuma(5), Norikazu Kiguchi(6), Satoshi Asano(1,2)
1. Dept of Cell Mol Pharmacol, Grad Sch of Biomed Health Sci, Hiroshima Univ, Hiroshima, Japan, 2. Sch of Dent, Hiroshima Univ, Hiroshima, Japan, 3. Dept of Dental Anesthesiol, Grad Sch of Biomed Health Sci, Hiroshima Univ, Hiroshima, Japan, 4. Dept of Pharmacol, Grad Sch of Biomed Health Sci, Hiroshima Univ, Hiroshima, Japan, 5. Dept of Pharmacol, Grad Sch of Dent, Osaka Univ, Osaka, Japan, 6. Dept of Physiol Sci, Sch of Pharmaceut Sci, Wakayama Med Univ, Wakayama, Japan
Keyword: AUTISM SPECTRUM DISORDER, PAIN, MICROGLIA, SPINAL CORD
One of the symptoms of autism spectrum disorder (ASD) under the DSM-5 is sensory abnormality such as hypersensitivity or hyposensitivity to sensory input. Individuals with ASD might display tactile abnormalities and/or abnormal emotional responses to tactile stimulation, but it remains unclear whether underlying sensory mechanisms are altered, or it is the emotional response to sensory input that leads to issues in filtering of the signal resulting in hyper/hypo-responsiveness. Emerging evidence suggests that microglia has a crucial role in forming and refining neuronal circuitry and network connectivity, and thus contributing to neuronal plasticity. In the present study, we have investigated the tactile sensitivity in a prenatal valproic acid (VPA)-induced model of ASD and subsequently assessed the possible involvement of microglial signaling in pain processing. Pregnant ICR mice were intraperitoneally injected with either VPA (500 mg/kg) or saline on embryonic day 12.5. Male offspring of VPA-treated mothers showed mechanical allodynia at both 4 and 8 weeks of age. In the dorsal horn of the spinal cord in prenatal VPA-treated mice, the numbers and staining intensities of Iba1-positive cells were increased and the cell bodies became enlarged, indicating the microglial activation. Administration of PLX3397, a colony-stimulating factor 1 receptor inhibitor, resulted in a decreased number of spinal microglia and attenuated mechanical allodynia in prenatal VPA-treated mice. Additionally, intrathecal injection of Mac1-SAP, a saporin-conjugated anti-CD11b antibody to deplete microglia, abolished mechanical allodynia in prenatal VPA-treated mice. These findings suggest that prenatal VPA treatment causes allodynia and spinal microglial activation might contribute to the increased nociceptive responses. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-189
ASDモデルマウスにおけるオキシトシン神経の神経回路と機能
Neural circuit organization and functions of oxytocin neurons in a mouse ASD model
*鶴谷 雅文(1,2)、宮道 和成(1)
1. 理化学研究所生命機能科学研究センター、2. 京都大学生命科学研究科
*Masafumi Tsurutani(1,2), Kazunari Miyamichi(1)
1. RIKEN Center for Biosystems Dynamics Research, 2. Graduate School of BIOSTUDIES,Kyoto University
Keyword: oxytocin, ASD, neural circuit, chemogenetic activation
Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder that is diagnosed in early childhood and is characterized by a high diagnosis rate of 1–2%. ASD is known to include the core symptoms of impaired social communication, repetitive behaviors, and limited interests, as well as a variety of peripheral symptoms. As the heritability of ASD is about 50%, environmental factors, especially early-life stage signals, must substantially contribute to the development of ASD. For example, exposure to antiepileptic agent valproic acid (VPA) during pregnancy is known to increase the ASD risk of children, which has been modeled in rodents. However, how and wherein such environmental factors affect developmental processes of social behaviors remain mostly unclear. Genetic association studies in human ASD patients and rodents ASD models both suggest the involvement of neural hormone oxytocin (OT) in the hypothalamus, as OT centrally controls social affiliative behaviors. We, therefore, aimed to characterize the neural circuit organization and functions of OT neurons in a VPA-induced ASD model. We found that VPA-induced social abnormality of mice was associated with a drastic reduction of OT immunostaining level in the hypothalamus. Notably, this reduction was not due to a loss of OT neurons, because VPA-treated mice harbored a comparable number of OT-Cre-based genetic labeling of OT neurons. These data suggest that OT protein expression is significantly reduced in each OT neuron of VPA-treated mice. Notably, chemogenetic activation of OT neurons not only restored social defects but also the expression level of OT protein of VPA-treated mice. By using rabies virus-mediated retrograde trans-synaptic tracing, we also found that the overall input connections to OT neurons were reduced in VPA-treated mice. Taken together, our data reveal multiple malfunctions developed in the OT system of VPA-treated mice, which may underlie the social defects. Future studies should reveal if these abnormalities commonly appear in other ASD models in mice and primates. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-190
行動テストバッテリーによるティモシー症候群新規病態モデルマウスの表現型解析
Phenotypic characterization of a novel mouse model for Timothy syndrome by a behavioral assessment battery
*潘 淼(1,2)、小澤 享弘(1,2,3)、上田 修平(1,2)、阿部 学(4)、崎村 健司 (4)、堀金 慎一郎(1,2)、竹本ー木村 さやか(1,2)
1. 名古屋大学環境医学研究所神経系分野Ⅰ、2. 名古屋大学院医学研究科分子神経科学、3. 名古屋大学院臓器病態診断学、4. 新潟大学脳研究所モデル動物開発分野
*Miao Pan(1,2), Yukihiro Ozawa(1,2,3), Shuhei Ueda(1,2), Manabu Abe(4), Kenji Sakimura(4), Shin-ichiro Horigane(1,2), Sayaka Takemoto-Kimura(1,2)
1. Dept. of Neuroscience I, Research Inst. of Environmental Med, Nagoya Univ, Nagoya, Japan, 2. Molecular/Cellular Neuroscience, Nagoya Univ. Grad. Sch. of Med., Nagoya. Japan, 3. Dept. of Pathology and Lab. Med., Nagoya Univ. Hospital, Nagoya, Japan, 4. Dept. of Comparative and Experimental Med., Brain Research Inst., Niigata Univ., Niigata, Japan
Keyword: autism spectrum disorder, Timothy syndrome
Timothy syndrome (TS), a multisystem developmental disorder characterized by autism spectrum disorders (ASD), syndactyly, and heart QT prolongation, is caused by a single de novo missense mutation G406R in L-type Ca2+ channel Cav1.2. This mutation causes impaired voltage-dependent channel inactivation and leads to excessive Ca2+ influx into cells (Splawski et al., 2004, 2005). The pathophysiological mechanism of this mutation remains largely unknown despite the great interest in causal relationships between the Ca2+ signaling abnormality and neurodevelopmental disorders. A TS mouse model, a heterozygous mouse with an inverted neomycin cassette in exon 8A (TS2-neo), was developed and TS2-neo was shown to exhibit ASD-related behaviors (Bader et al., 2011, Horigane et al., 2020). However, TS2-neo mice showed low expression level of G406R mutant channels and general knock-in of the mutation make it difficult to identify the outcome of G406R mutant channels expressed in the nervous system. Therefore, we have developed a nervous system-specific G406R knock-in mouse with physiological expression levels of mutant channels in the central nervous system introduced by a Cre-recombinase-dependent system. In this study, we conducted a series of behavioral assessments on this novel conditional knock-in (cKI) mouse line. We observed that the cKI mice displayed disability in motor coordination in the rotarod test related to the impairment of gross motor skills exhibited by patients with TS. ASD-related behaviors associated with the core triad of ASD were reproduced in the cKI mice, with a more repeated behavior in the grooming test and less social interaction time in the three-chamber test. Furthermore, cKI mice also displayed hyperactivity in the open field test and an abnormal fear memory in the fear conditioning test. Thus, we developed a novel TS mouse model bearing construct and face validity, which help us understand the complex pathophysiology of TS associated with ASD induced by abnormal Ca2+ signaling in the nervous system. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-191
自閉スペクトラム症者における恐怖関連刺激による視覚時間順序判断低下の神経相関
Neural correlates of the impaired visual temporal order judgment by fear-relevant stimulus in individuals with autism spectrum disorders
*渥美 剛史(1,2)、井手 正和(2)、Chakrabarty Mrinmoy(3)、寺尾 安生(1)
1. 杏林大学、2. 国立障害者リハビリテーションセンター研究所、3. Indraprastha Institute of Information Technology Delhi
*Takeshi Atsumi(1,2), Masakazu Ide(2), Mrinmoy Chakrabarty(3), Yasuo Terao(1)
1. Kyorin University, 2. Research Institute of National Rehabilitation Center for Persons with Disabilities, 3. Indraprastha Institute of Information Technology Delhi
Keyword: autism spectrum disorder, anxiety, temporal order judgment, fMRI
Atypical sensory features in individuals with autism spectrum disorder (ASD) and a high co-morbidity with anxiety disorder are gaining more attention in recent years. Potential linkages between them have been frequently suggested (MacLennan et al., 2020; Pickard et al., 2020), but the underlying mechanism is still unknown. We previously reported that negative emotion signals modulate the precision of temporal order judgment (TOJ) in ASD (Chakrabarty et al., 2021). The present functional magnetic resonance (fMRI) study examined neural correlates of emotion-induced regulation of sensory temporal processing in ASD. We asked 11 ASD and 8 TD participants to perform the TOJ task; they were required to answer the temporally later presented side (upper/lower) of the two successive white circles presented at the left peripheral field of a monitor screen. Participants performed the task during the anatomical scan and the just noticeable difference (75% threshold) was calculated from the response data at various stimulus onset asynchronies (SOA). Based on the individual threshold, participants underwent fMRI of the TOJ at one of the SOAs. In this scan, a face image expressing fear (FE) or neutral (NE) appeared at the center of the screen prior to the two stimuli for the TOJ. We analyzed correct response rates and fMRI signals during the TOJ in each face condition. An analysis of variance revealed a significant interaction between the diagnostic group and face condition (F = 4.75, p = 0.04, partial η2 = 0.22). A post-hoc analysis showed a greater reduction of correct responses during FE trials in ASD than in TD group (p = 0.03). A whole-brain analysis (FE > NE and resting states contrast, pFWE-corr < 0.05) revealed greater signals in the right precentral, paracingulate gyri, and the left fusiform cortex in TD than in ASD. A multiple regression analysis with individual differences of FE and NE correct response rate for ASD group showed associations with the right middle frontal gyrus, precuneus cortex, and the bilateral cerebellum. The results from the ongoing study suggest greater fear-induced responses in ASD. We speculate an enhancement of freezing response mediated by the parasympathetic nervous system to fear-relevant signals (Roelofs, 2017) deactivated the internal timing processes. It seems that this response was induced by an altered modulatory function in emotion-related circuits, including the cingulate cortex and the fusiform-amygdala systems (Dziobek et al., 2010). Performance changes in ASD might be mediated by altered activities in the TOJ-related regions (Miyazaki et al., 2016). Our findings may imply a neural foundation of the modulatory role of emotion-induced over-reactivity for the temporal processing of sensory stimuli in ASD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-192
胎生期の化学物質投与は小脳小葉の過形成と顆粒細胞移動の異常を誘発する
Prenatal chemical exposures induce the hyperplasia of lobules in developing cerebellar cortex and defection of granule cell migration
*吉田 祥子(1)、Veloo Sharumadhi(1)、諫田 泰成(2)
1. 豊橋技術科学大学、2. 国立医薬品食品衛生研究所
*Sachiko Yoshida(1), Sharumadhi Veloo(1), Yasunari Kanda(2)
1. Toyohashi University of Technology, 2. National Institute of Health Sciences
Keyword: developmental neurotoxicity, cerebellum, granule cell, hyperplasia
Chemical exposure in utero has potential effects on developmental neurotoxicity (DNT). Multiple chemicals, for example, sodium valproate (VPA), chlorpyrifos (CPF), or lipopolysaccharide (LPS), have been associated with an increased risk of ASD. We have observed that VPA-administrated rats showed excess folding, hyperplasia between the V to VI lobules of cerebellar vermis within two weeks after birth with dose-dependent and administrated-period-dependent manner. Cerebellar excess development is maintained in adults, and its alteration is similar to the human early ASD cerebellum. The irregular folding of vermis appeared in butyrate (BA)- and LPS-administrated rat cerebellum in early weeks, but not in CPF-administrated one. This irregular folding of lobules appeared even in the P5 cerebellum of the VPA-administrated rat. Recently, some reports presented that the selective knockdown of liver kinase B1 (LKB1), an activator of AMPK, induced cerebellar hyperplasia. In the VPA- or BA-administrated cerebellum, LKB1 expression disappeared in the inner EGL, whereas LKB1 was expressed in the control. In the VPA-, BA-, or LPS-administrated cerebellum, the granule cell migration sometimes defected, and the expressions of H3K9me were decreased in the molecular layer. We suggest that the epigenetic alteration of developing cerebellum with chemical exposure would induce the aberrant differentiation of granule cells, preventing the pruning of irregular neuronal development to hyperplasia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-193
マーモセット自閉症モデルとヒト自閉症サブタイプおよび15q重複症候群のトランスクリプトームの類似性は遺伝因子と環境因子の共通メカニズムを示唆する
Transcriptomic similarity of marmoset autism model and human autism subtypes/15q duplication syndrome suggests convergent mechanisms for genetic and environmental factors
*渡邉 惠(1)、小賀 智文(1)、中垣 慶子(1)、一戸 紀孝(1)
1. 国立精神・神経医療研究センター
*Satoshi Watanabe(1), Tomofumi Oga(1), Keiko Nakagaki(1), Noritaka Ichinohe(1)
1. National Center of Neurology and Psychiatry
Keyword: AUTISM SUBTYPES, VALPROIC ACID, TRANSLOCATOR PROTEIN
Autism spectrum disorder (ASD) is characterized by impaired social interaction and communication, restricted interests, and repetitive behaviors. ASD is considered a heterogeneous disorder with both genetic and environmental factors. Stratification of ASD and comparison with animal models will be needed for a better understanding of the disease. Marmosets exposed to valproic acid (VPA) in utero develop behavioral and synaptic abnormalities similar to human ASD (Watanabe et al. Nat. Commun. 2021). However, the correlation of cortical gene expression modulations between the entire human ASD population and the marmoset model appeared to be modest, which is presumably due to the heterogeneous nature of ASD. We applied hierarchical clustering to transcriptome data from the postmortem human cortex (Parikshak et al. Nature 2016) and revealed three groups of ASD individuals. Group 1 (26%) showed an enhanced expression of genes in co-expression modules associated with microglia and astrocytes. Group 2 (18%) showed a reduced expression in modules associated with neurons, in addition to enhanced expression in microglia- and astrocyte-related modules. Group 3 (56%) did not show modulations in any of these modules. People with chromosome 15q duplication were prevalent in groups 1 and 2. The gene expression modulations in the marmoset model were highly correlated with those in human groups 1 (r = 0.67) and 2 (r = 0.60), as well as those with 15q duplication (r = 0.72), but not with those in group 3 (r = -0.37). Expression of translocator protein (TSPO) gene, which is a marker of activated microglia, was also elevated in human groups 1 and 2, with a high diagnostic ability as suggested by the ROC analysis (AUC = 0.81). Expression of TSPO in the marmoset model was also correlated with those in human groups 1 and 2, but not in group 3. These results suggest that the VPA-exposed marmoset model replicates a subset of ASD people with abnormal glial and neuronal conditions in the cortex. The high correlation between the VPA model and 15q duplication syndrome is surprising because it suggests that genetic and environmental factors cause convergent gene expression modulations in the autistic brain. This may be related to epigenetic effects of VPA. Biomarkers for the modulated genes, such as TSPO PET scan, may be used to detect people with these phenotypes, and the marmoset model may help to understand the pathology and to develop medication for those people. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-194
ASDにおける感覚異常の神経回路基盤の解明
Elucidating the neural circuit basis of sensory abnormalities in ASD.
*渋下 碧(1)、竹田 育子(1,2)、加藤 大輔(1,2)、森 大輔(3)、久島 周(3)、尾崎 紀夫(3)、和氣 弘明(1,2)
1. 名古屋大学大学院医学系研究科 分子細胞学、2. 生理学研究所 多細胞回路動態研究部門、3. 名古屋大学大学院医学系研究科 精神医学
*Midori Shibushita(1), Ikuko Takeda(1,2), Daisuke Kato(1,2), Daisuke Mori(3), Itaru Kushima(3), Norio Ozaki(3), Hiroaki Wake(1,2)
1. Department of Anatomy and Molecular Cell Biology, Nagoya University Graduate school of Medicine, 2. Division of Multicellular Circuit Dynamics, National Institute for Physiological Sciences, 3. Department of Psychiatry, Nagoya University Graduate school of Medicine
Keyword: ASD, sensory abnormalities, neural activity
Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder with impaired social interactions and communications, limited interests, and repetitive behaviors. ASD is caused by abnormal neuronal circuit formation due to genetic and environmental factors such as maternal inflammation. One notable symptom in ASD is sensory abnormalities, with about 90 % of patients reported to have atypical sensory experiences. Although increased functional connectivity of thalamus-sensory cortex and hyperactivity of S1 and amygdala in response to sensory stimulation have been reported, the pathological basis of neuronal circuitry that causes sensory abnormalities remains unknown. Furthermore, the pathological reciprocal interaction between sensory abnormalities and impaired social behavior has not been demonstrated. In this study, we used two ASD model mice, maternal immune activation (MIA) with Poly(I:C) mice and 3q29 deletion mice lacked a site on chromosome including risk genes of ASD. To study the functional sensory evoked response of neuronal population and their contribution on functional local circuit connectivity in ASD model mice, we visualized neuronal activities with whisker stimulation or with holographic single cell stimulation in the primary somatosensory cortex barrel field (S1BF) using in vivo two-photon holographic microscopy combined with virus targeted fluorescent expression. Whisker-response cell population, which showed high correlation with whisker stimuli, in S1BF of ASD model mice showed lower synchrony, and higher failure rate for 10 serial stimuli, compared to control. The results suggest the variance of neuronal responses to whisker stimuli, which may lead to increased variability in the expression of higher brain functions in ASD. In addition, we examined functional connectivity changes in S1BF local neuronal circuits and input changes from other regions to S1BF using holographic stimulation. ASD model mice showed increased functional connectivity in S1BF, which may interfere with normal responses to whisker stimuli by altering neuronal reciprocal to sensory input of ASD. This study provides the pathological perspective of sensory abnormalities and their abnormal circuitry-basis from both genetic and environmental factors of ASD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-195
母親や子のFABP4機能低下が自閉症病態形成に果たす役割の検討
Hypofunction of FABP4 in mothers or pups in the pathophysiology of autism spectrum disorder
*前川 素子(1)、孫 正康(1)、大西 哲生(2)、吉川 武男(3)、大和田 祐二(1)
1. 東北大学、2. 東京医科歯科大学、3. 理化学研究所
*Motoko Maekawa(1), Zhengkang Sun(1), Tetsuo Ohnishi(2), Takeo Yoshikawa(3), Yuji Owada(1)
1. Tohoku University, 2. Tokyo Medical and Dental University, 3. RIKEN
Keyword: FABP4, autism spectrum disorder, metabolism, inflammation
Autism spectrum disorder (ASD) is a neurodevelopmental disorder characterized by difficulties in social communication and interaction, as well as repetitive and characteristic patterns of behavior. The pathogenesis of ASD remains still largely unknown. However, being overweight or obese during infancy and low weight at birth are known as risks, suggesting a metabolic aspect. Fatty acid binding protein 4 (FABP4), which functions as an intracellular chaperone for hydrophobic molecules such as long-chain fatty acids and an adipokine secreted from the adipose tissue, is known to be one of the key molecules associated with obesity. In our previous study, we identified two novel and functional rare variants of the FABP4 gene in ASD. One of them was a nonsense mutation, thereby disrupting the gene function. We also indicated that Fabp4 homozygous knockout (KO) mice displayed ASD-relevant behavioral and histological phenotypes. Therefore, it is likely that the hypofunction of FABP4 in children during the embryonic and/or postnatal stages may contribute to the pathophysiology of ASD. In addition to this simple interpretation of the findings, we came up with the possibility that maternal deficit in FABP4 function could also contribute to the pathogenesis of ASD. This idea is supported by the following observations: 1) the above-stated FABP4 nonsense mutation was transmitted from the mother, who developed depression, to the ASD proband, and 2) heterozygous females were being used to produce homozygous Fabp4 KO mice. Furthermore, it is known that FABP4 hypofunction in mothers can alter the maternal metabolic status and inflammatory cytokine levels. Collectively, we assume that neurodevelopment of fetuses and/or newborns is potentially affected when FABP4 hypofunction occurs in the mother’s body. In the current study, we aimed to experimentally test this idea by administering a synthetic FABP4 inhibitor, BMS309403, to mice. We treated inbred C57BL6/J dams, their pups or both, with BMS309403 or vehicle, setting four experimental groups: (1) mothers_vehicle + pups_vehicle, (2) mothers_vehicle + pups_ BMS309403, (3) mothers_MS309403 + pups_vehicle, and (4) mothers_BMS309403 + pups_BMS309403. First, we examined whether the treatments affected the levels of metabolites and pro-inflammatory cytokines in the serum of dams and pups to monitor metabolism and inflammation status in the body. Second, to examine whether the pups from each experimental group exhibit ASD-like phenotypes, we conducted behavioral testing, which includes ultrasound vocalization and homing behavior tests, and morphological analyses of dendritic spines of pyramidal neurons in the brain. In this presentation, we show those results and discuss the potential roles of FABP4 hypofunction in mothers and children in the pathophysiology of ASD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-196
偶発性自閉症モデルであるBTBR T+ltpr3tf/Jマウスの自閉症様行動を制御する神経回路の検討
Dissecting neural circuits responsible for autism-like behavior in BTBR T+ltpr3tf/J mice as an idiopathic model of autism.
*荒川 礼行(1)、樋口 裕城(1)、朔晦 峻一(2)、三治 篤主(2)、仲地 泰雅(2)
1. 琉球大学医学研究科、2. 琉球大学医学部
*Hiroyuki Arakawa(1), Yuki Higuchi(1), Shunichi Tachigori(2), Atsuto Sanji(2), Taiga Nakachi(2)
1. University of the Ryukyus School of Medicine, Okinawa, Japan, 2. University of the Ryukyus Dept Medicine, Okinawa, Japan
Keyword: AUTISM, SOCIAL DEFICITS, NEURAL CIRCUITS, BTBR
Autism spectrum disorders (ASD) are characterized by social-communicative behavior impairments and persistent repetitive behaviors. Both of these behavioral phenotypes are displayed in preclinical animal models, BTBR T+ltpr3 tf/J (BTBR) mouse. Using genetical and pharmacological tools, dissection of these behaviors and underlying neural mechanisms in BTBR mice will provide discrete features of their neural mechanisms controlling specific behavioral phenotypes relevant to autism symptoms. We used a combination of behavioral, morphological, and physiological assessments and dissected out the neural circuits responsible for the expression of specific behaviors relevant to autism symptoms. In comparison of BTBR mice with C57BL/6J mice (B6) as a social standard strain, recognition of social relationships between conspecifics was measured by social signaling behavior, scent marking toward appropriate social situations. Social approach as a sociability measurement and excessive grooming behavior as an obsessive repetitive behavior are also assessed as a function of responses to specific social situations. In addition, serotonergic neural circuits as a primary regulator for these autism-like behaviors are characterized in these strains using neural tracer and immunohistochemical analysis. These multiple analyses of behavioral neural circuits in BTBR mice revealed that maladaptive-integration of neural circuits exhibited in BTBR mice produced differentiated socio-cognitive processes along with dysfunction of serotonergic neurons. The behavioral characteristics shown in BTBR mice are not solely a lack of functional neural circuits, but rather a mal-formation of neuronal pathways controlling adaptive behaviors. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-197
Brain-Behavior Dimensions Delineate Autism Spectrum Disorder Subgroups Linked to Distinct Molecular Pathways
*Buch Amanda(1)、Vertes Petra(2)、Seidlitz Jakob(3,4)、Kim So Hyun(1)、Grosenick Logan(1)、Liston Conor(1)
*Amanda M. Buch(1), Petra Vertes(2), Jakob Seidlitz(3,4), So Hyun Kim(1), Logan Grosenick(1), Conor Liston(1)
1. Dept Psychiatry and Brain and Mind Res. Inst., Weill Cornell Medicine, Cornell University, NY, USA, 2. Dept Psychiatry, Univ. of Cambridge, Cambridge, United Kingdom, 3. Dept Psychiatry, Univ. of Pennsylvania, Philadelphia, PA, 4. Child and Adolescent Psychiatry and Behavioral Sci., Children’s Hosp. of Philadelphia, Philadelphia, PA
Keyword: MACHINE LEARNING, GENE EXPRESSION, RESTING STATE FUNCTIONAL MRI, AUTISM SPECTRUM DISORDER
Biomarkers have transformed diagnosis and treatment in medicine, but are elusive in neuropsychiatric syndromes such as autism spectrum disorder (ASD), a heterogeneous neurodevelopmental disorder. The two core symptoms that define ASD are social communication impairments and repetitive and restricted behaviors and interests (RRB), and there is a wide range of cognitive and language abilities. While ASD is 80-85% heritable with over 100 common genetic risk variants and brain-region-specific transcriptional changes, it is not known how distinct molecular and network-level mechanisms give rise to the heterogeneous behavioral symptoms in individuals—a major barrier to developing effective treatment. Using a large, publicly available neuroimaging dataset comprising resting state functional magnetic resonance imaging (rsfMRI) scans from N=299 subjects with ASD and N=907 neurotypical controls, we identified three latent dimensions of functional brain network connectivity that predict individual differences in ASD symptoms and behaviors. We show that patients with ASD can be grouped into distinct neurophysiological subgroups based on patterns of dysfunctional connectivity and clinical behaviors. In this cohort, functional connectivity features were extracted from rsfMRI data, regularized canonical correlation analysis was used to identify associations between connectivity features and behavioral data, and ASD subjects were clustered along these dimensions. Cross-validation analyses showed high stability in the brain-behavior dimensions, with replicable clusters in held-out data. Next, we integrated neuroimaging data with gene expression data from the Allen Human Brain Atlas, and found that within each subgroup, ASD-related functional connectivity was explained by regional differences in the expression of distinct gene sets that were enriched for ASD-associated genes (SFARI database) and different molecular associations. In sum, our results identify discrete ASD subgroups associated with specific ASD behaviors and neurophysiological signatures, and these different forms of ASD implicate distinct genetic mechanisms. The results of this study suggest a promising new approach for understanding the neurobiological substrates of ASD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-198
電位依存性L型カルシウムチャネルにおける機能獲得型変異に伴うティモシー症候群の中枢神経特異的新規モデルマウスの樹立
Development of a nervous system-specific conditional knock-in mouse strain of a gain-of-function mutation in L-type Ca(2+) channel associated with Timothy syndrome
*小澤 享弘(1,2,3)、潘 淼(1,2)、阿部 学(4)、崎村 健司(4)、堀⾦ 慎⼀郎(1,2)、⽵本ー⽊村 さやか(1,2)
1. 名古屋大学環境医学研究所神経系分野I、2. 名古屋大学⼤学院医学系研究科分⼦神経科学、3. 名古屋大学⼤学院医学系研究科臓器病態診断学、4. 新潟⼤学脳研究所モデル動物開発分野
*Yukihiro Ozawa(1,2,3), Miao Pan(1,2), Manabu Abe(4), Kenji Sakimura(4), Shin-ichiro Horigane(1,2), Sayaka Takemoto-Kimura(1,2)
1. Dept. of Neuroscience I, Research Inst. of Environmental Med., Nagoya Univ., Nagoya, Japan, 2. Molecular/Cellular Neuroscience, Nagoya Univ. Grad. Sch. of Med., Nagoya. Japan, 3. Dept. of Pathology and Lab. Med., Nagoya Univ. Hospital, Nagoya, Japan, 4. Dept. of Comparative and Experimental Med., Brain Research Inst., Niigata Univ., Niigata, Japan
Keyword: AUTISM SPECTRUM DISORDER, TIMOTHY SYNDROME
Ca2+ signaling plays a critical role in multiple cellular functions, including neural circuit formation and plasticity. Pathologically, the dysfunction of Ca2+ signaling is linked with psychiatric and neurological disorders, including autism spectrum disorder (ASD). Timothy syndrome (TS) is a rare systemic disorder caused by a gain-of-function mutation, G406R, in L-type Ca2+ channel (Cav1.2), causing excessive Ca2+ influx into the cells (Splawski et al., 2004, 2005). TS is characterized by ASD, gross motor dysfunction, and heart QT prolongation. However, its underlying pathophysiological mechanisms remain largely unknown. To elucidate the impact of the mutant channel in the central nervous system during development, we focused on the migration of immature neurons. Our work on the TS mouse model, TS2-neo, a previously established G406R knock-in mouse strain, revealed abnormal migration of the inhibitory neurons (Bader et al., 2011, Horigane et al., 2020). Several research groups, including ours, have reported ASD-related behaviors and neurodevelopmental structural abnormalities in the TS2-neo mouse model, and these findings significantly contributed to the understanding of the TS pathology. However, it should be noted that this mouse model is a global knock-in mouse and the expression levels of G406R mutant channels are suppressed to avoid mortality before weaning. Consistent with the previous studies, we confirmed low expression levels of transcripts encoding G406R mutant channels leading to lower expression levels of total transcripts of Cav1.2 in TS2-neo.To assess the effect of G406R mutant channels at a physiological expression level on the nervous system, we developed a new TS model mouse strain, nervous system-specific G406R knock-in mouse utilizing the Cre-recombinase system. We confirmed higher levels of mutant channels in the nervous system in the conditional knock-in mice compared with TS2-neo. Interestingly, a nervous system-specific G406R knock-in mouse strain showed lower body weight than the wild-type littermates (measured between 3–8 weeks of age) indicating systemic growth impairment also reported in patients with TS. We also observed some histological differences in the neocortex of the mutant mouse strain, which may relate to the abnormal migration of the inhibitory neurons during development. Together with the behavioral analysis (presented in another poster by Pan et al.), we successfully developed the conditional knock-in mouse strain as a novel mouse model for TS. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-199
自閉スペクトラム症マーモセットモデルにおけるオキシトシン経鼻投与効果の可能性
Prospective effect of nasal oxytocin administration in a marmoset model of autism
*中村 月香(1)、中神 明子(3)、中垣 慶子(1)、土屋 明子(1)、川合 伸幸(2)、一戸 紀孝(1)
1. 国立精神・神経医療研究センター神経研究所微細構造研究部、2. 名古屋大学大学院情報学研究科、3. 日本女子大学人間社会学部心理学科
*Madoka Nakamura(1), Akiko Nakagami(3), Keiko Nakagaki(1), Akiko Tsuchiya(1), Nobuyuki Kawai(2), Noritaka Ichinohe(1)
1. Dept. Ultrastructural Research, NCNP, Tokyo, Japan, 2. Dept. Cognitive and Psychol Sciences, Nagoya Univ, Aichi, Japan, 3. Dept. Psychol, Japan Women's Univ, Kanagawa, Japan
Keyword: AUTISM SPECTRUM DISORDER, OXYTOCIN
Social interactions are characterized by a series of other-regarding decisions and coordinated actions. People with autism spectrum disorders (ASD) have a complex set of problems in social interactions that are unique to each patient. It is therefore necessary for autism research to establish a broad-spectrum assay covered not only simple biomarkers but also higher social cognition skills. We have previously demonstrated that the New World common marmoset monkeys (Callithrix jacchus) react to inequity if they witness a conspecific that obtains a more attractive reward for the same effort, whereas the valproic acid-induced marmoset model of ASD do not. In this study, we tested whether this autistic phenotype in the non-human primate model of ASD can be restored with a neurohypophysial hormone, oxytocin (OT). The behavioral task employed in this study required holding a plastic spoon for 2 seconds. Adult male autistic marmosets received a nasal dose of either OT or saline 30 minutes before each spoon-holding test session. We found that the autistic marmosets who received OT tended to avoid inequity than when they were given saline before the test. This study confirms that the nasal administration of oxytocin may be effective in improving higher-order social skills. Although this is an ongoing study and a larger sample size is needed, this result indicates that the behavioral phenotype of inequity-aversion tendency in the marmoset model of autism can be used as an assay to develop treatments for higher-order social deficits faced by individuals with ASD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-200
パーキンソン病モデルマウスにおけるシヌクレイン病理に対するミクログリア介在性神経炎症の影響
Aging-induced microglia-mediated neuroinflammation exacerbates α-synucleinopathy in Parkinson disease model mice
*佐木山 裕史(1)、早川 英規(1)、木村 康義(1)、Aguirre Cesar(1)、池中 建介(1)、長野 清一(1)、馬場 孝輔(1,2)、望月 秀樹(1)
1. 大阪大学 、2. 富山大学
*Hiroshi Sakiyama(1), Hideki Hayakawa(1), Yasuyoshi Kimura(1), Cesar Aguirre(1), Kensuke Ikenaka(1), Seiichi Nagano(1), Kosuke Baba(1,2), Hideki Mochizuki(1)
1. Osaka University, 2. University of Toyama
Keyword: Parkinson's disease , neuroinflammation, Lewy pathology, microglia
Parkinson's disease (PD) is a neurodegenerative disorder pathologically characterized by the accumulation of intracytoplasmic Lewy bodies and progressive degeneration of dopaminergic neurons in the substantia nigra (SN). α-synuclein, a 140-amino acid protein, is the major filamentous component of Lewy bodies. Several findings in animal models have implicated cell-to-cell propagation of α-synuclein, which seed aggregation of endogenous α-synuclein in recipient cells resulting in dopaminergic neuronal loss, as a significant contributor in PD pathology. Intra-parenchymal inoculation of exogenous α-synuclein, mainly α-synuclein fibrils, is one of the most common animal models used for studying α-synuclein accumulation, aggregation and propagation. On the other hand, previous clinical and experimental research suggests that neuroinflammation, an inflammatory response within the central nervous system, plays a central role in the dopaminergic neuronal loss in PD. Some studies showed that overactivated microglia induce severe neurotoxic effects and neuroinflammation in PD. In addition, aging is a known risk factor in PD and many studies show that aging disrupts the control of microglia-mediated inflammation. In this study, we evaluated the effect of microglia-mediated neuroinflammation on PD pathology using α-synuclein fibrils-inoculated senescence accelerated mice. The senescence accelerated mouse prone 8 (SAMP8) is a widely used rodent model of aging and senile dementia, while the senescence accelerated mouse resistant 1 (SAMR1) that does not show these senescence-related phenotypes is commonly used as control. We injected α-synuclein fibrils into the SN of SAMP8 mice (N=10) and the control group, SAMR1 mice (N=13). At 24 weeks after fibril injection, we assessed PD pathology and dopaminergic neuronal loss in the SN of both groups by immunohistochemical analyses. We observed that SAMP8 mice showed more widespread PD pathology and progressive degeneration of dopaminergic neurons in the SN compared to SAMR1 control mice. We further found that the number of Iba1-positive cells, indicative of activated microglia, in the SN of SAMP8 mice increased compared to that of SAMR1 mice. These findings suggest that microglia-mediated neuroinflammation plays an important role in PD pathology and aging exacerbates these processes. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-201
前駆期パーキンソン病モデルマウスはミトコンドリア障害を示す～血漿メタボローム解析の結果から～
Mitochondrial dysfunction in a mouse model of prodromal Parkinson’s disease: metabolomic analysis
*生野 真嗣(1)、山門 穂高(1)、髙橋 良輔(1)
1. 京都大学大学院医学研究科 臨床神経学
*Masashi Ikuno(1), Hodaka Yamakado(1), Ryosuke Takahashi(1)
1. Department of Neurology Kyoto University Graduate School of Medicine
Keyword: Parkinson's disease, alpha synuclein, animal model, metabolomic analysis
[Objective] For the development of disease-modifying therapies for Parkinson’s disease (PD) the identification of biomarkers in the prodromal stage is urgently required. Because PD is considered a systemic disease even in the early stage, we performed a metabolomic analysis of the plasma from a mouse model of prodromal PD (p-PD). [Methods] In the present study, we performed metabolomic analysis of plasma from a mouse model of prodromal PD. This mouse model was created based on two major genetic risk factors of idiopathic PD by crossing α-syn Bacterial Artificial Chromosome (BAC) transgenic mice with heterozygous knockout of Gba1 gene that encodes beta-glucosidase (GCase). These mice express human α-syn in a physiological manner through its native promoter and showed mild dopaminergic cell loss coupled with α-syn accumulation in PD-vulnerable regions without motor phenotypes, suggesting being a prodromal model of PD. For metabolomic analysis, blood plasma was collected from 2 or 3 mice as one sample. 5 samples for each group were prepared for analysis in capillary electrophoresis - time-of-flight mass spectrometry (CE-TOFMS) and liquid chromatography- time-of-flight mass spectrometry (LC-TOFMS). [Results] Total of 314 metabolites were analyzed, and among them 10 were judged as significantly changed (P<0.05). From the viewpoint of p-value (P<0.05), the value ratio of two groups (>1.2 or <0.8) and identifiable peak of all the sample of the mice in mass spectrometry, we picked up isobutyrylcarnitine and pyrimidine nucleotides (thymidine and deoxyuridine) in the important results. Increased levels of isobutyrylcarnitine in p-PD mice imply an abnormality in β-oxidation in mitochondria, and increased levels of pyrimidine nucleoside can be associated with mitochondrial dysfunction. Consistent with these results, the immunoblot analysis showed a defect in mitochondrial complex I assembly in p-PD mice. [Conclusions] These results suggest that systemic mitochondrial dysfunction may exist in p-PD mice and contribute to the pathogenesis of PD, potentially being useful as early biomarkers for PD. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-202
ADHおよびALDH2のSNP*におけるLRPおよびsmadの変異はOSTEOPOROSIS-PSEUDOGLIOMA SYNDROME (OPPG)の同変異に近似する
ABNORMAL LRP AND SMAD PATHWAY INDUCED OSTEOPOROSIS-PSEUDOGLIOMA SYNDROME (OPPG) LIKE GENETIC TYPE IN ADH- AND ALDH2-SNP*.
*渡邊 利明(1)、竹内 仁(1)、杉山 渉(1)、杉浦 加奈子(1)
1. 帝京科学大学
*Toshiaki Watanabe(1), Hitoshi Takeuchi(1), Wataru Sugiyama(1), Kanako Sugiura(1)
1. Teikyo University of Science
Keyword: OPPG, Wnt-signal, HRP, beta-catenin
1) Introduction and Background: We previously reported that Osteoporosis Factor Gene of ADH- and ALDH2-mutation caused abnormal turnover in Rank-RankL-OPG signaling. Thereby, the relationship between C/EBP and PPARγ was stabilized.
2) Aim: How and which in ADH- and ALDH2-gene polymorphisms (snp), had genic effect of Wnt (Lrp6/5 signaling) pathway and/or TGF-beta (Smad2/3 signaling) gene transcription like OPPG?
3) Method: Human study (n=240) was approved by an Institutional Ethical Review Board (Teikyo Univ. of Sci.). Extraction, PCR and RT-PCR amplification and sequencing of mitochondrial DNA and mRNA from human hair root cell were used by MyGo Pro/MyGo Mini®.
4) Result: In ADH-snp, the change of ratio in LRP5/LRP6-, in RankL /Rank-, in TGF-beta-expression has shown, and induced widely fluctuation of turnover in smad1/smad6. More over in ALDH2-snp*, wnt16-gene and beta-catenin expression were changed. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-203
α-Synuclein fibrils induced disruption of pacemaker firing in dopamine neurons is dependent on selective K-ATP channel activation
*Thakur Poonam(1)、Roeper Jochen (2)
*Poonam Thakur(1), Jochen Roeper(2)
1. School of Biology, IISER Thiruvananthapuram, Kerala, India, 2. Institute for Neurophysiology, Goethe University, Frankfurt, Germany
Keyword: α-synuclein , ion-channel, substantia nigra
Parkinson’s disease is associated with α-synuclein aggregation and progressive loss of substantia nigra dopaminergic neurons (SN DA-neurons). Precise mechanism(s) behind selective vulnerability of SN DA-neurons to α-synuclein aggregation in comparison to ventral tegmental area (VTA) DA neurons is not known. Even within SN, DA-neurons in ventro-lateral tier degenerate earlier and to a larger extent compared to the neurons in dorso-medial tier. Present study is aimed to delineate the basic of their differential vulnerability to fibrillar α-synuclein. Male C57Bl/6N mice were injected with red fluorescence beads in nucleus accumbens or dorso-medial striatum or dorso-lateral striatum that retrogradely travel to VTA, medial-SN and lateral-SN respectively. After 3-4 days of the injection, acute mid-brain slices were prepared and autonomous pacemaker firing of labelled neurons was studied in presence of fibrillar or monomeric α-synuclein. Nanomolar concentrations of α-synuclein fibrils, but not its monomeric forms, selectively and acutely disrupted pacemaker firing of lateral-SN DA neurons. Their firing became irregular and slow before stopping eventually. On the other hand, firing in medial SN or VTA was unaffected. Further, pharmacological inhibition or genetic ablation of K-ATP channels in the lateral-SN DA neurons protected from α-synuclein fibrils induced disruption of their firing proving their role in mediating this effect. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-204
Modulation of liquid-liquid phase separation behavior of Fused in sarcoma (FUS) using antisense oligos
*Mohammad Moniruzzaman(1), Yusuke Fujioka (1), Akihide Takeuchi(2), Akio Masuda (3), Masahiro Neya (4), Shinsuke Ishigaki(1)
1. Dept of Neurology, Nagoya Univ Grad Sch of Medicine, Aichi, Japan, 2. Dept of Developmental Biol and Functional Genomic, Ehime Univ Grad Sch of Medicine, Ehime, Japan, 3. Division of Neurogenetics, Center for Neurological Dis and Cancer, Nagoya Univ Grad Sch of Medicine, Aichi, Japan, 4. KNC Laboratories Co., Ltd., Hyogo, Japan
Keyword: FUS , Liquid-liquid phase seperation, Antisense oligo, FUS-ALS/FTLD
Background: Liquid-liquid phase separation (LLPS) of proteins, formation of condensed liquid droplets in a dilute bulk phase, is involved in diverse cellular processes such as transcription, splicing, DNA condensation, and DNA repair. RNA binding protein Fused in sarcoma (FUS) forms liquid droplets that can transform into hydrogel and amyloid fibrils resulting in neurodegenerative diseases including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar degeneration (FTLD). Mounting evidence suggested that phase separation of FUS is crucially involved in ALS/FTLD pathogenesis and it was reported that the presence of RNA could ameliorate its liquid droplet formation. These findings prompted us to develop antisense oligos (ASOs) can potentially modulate LLPS of FUS and serve as promising drug candidates against ALS/FTLD.
Objective: The aim of this research is to investigate the effect of FUS targeting ASOs on liquid-liquid phase separation behavior of FUS in vitro.
Methods: MBP-FUS R495X was purified by expressing in Escherichia coli BL21 DE3. Full length MBP-FUS R495X formed droplets when it was diluted. We monitored phase separation via morphological detection of droplet formation and in a turbidity assay, where optical density of the protein solution is used as a measure of phase separation. Candidate ASOs were designed based on the sequences and the structure of FUS-binding RNA. Screening of ASOs was performed in a turbidity assay and by observing the distribution of FUS R495X in NSC-34 cells upon ASO treatment.
Results: Screening data demonstrated that 100nm of ASO NKK-003, NKK-004, and NKK-009 reduced turbidity of MBP-FUS R495X solution after both 4h and overnight incubation. FUS R495X is known to associate with ALS and shows extensive cytoplasmic mislocalization. Consistent with turbidity assay, NKK-003, NKK-004, and NKK-005 reversed the localization of FUS R495X from the cytoplasm to the nucleus in NSC-34 cells.
Conclusion: Our in vitro and cell culture data revealed that FUS targeting ASOs can modulate FUS LLPS. Further investigation would provide valuable insight for the development of ASO based FUS-ALS/FTLD intervention. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-205
GBA1ノックアウトメダカを用いたGBA1関連αシヌクレイノパチーの疾患修飾薬の探索
The search for compounds to treat GBA1-related α-synucleinopathy using gba1 knock-out medaka
*中西 悦郎(1)、上村 紀仁(1)、澤村 正典(1)、樽野 陽亮(1)、山門 穂高(1)、髙橋 良輔(1)
1. 京都大学大学院医学研究科
*Etsuro Nakanishi(1), Norihito Uemura(1), Masanori Sawamura(1), Yosuke Taruno(1), Hodaka Yamakado(1), Ryosuke Takahashi(1)
1. Grad Sch Med, Univ of kyoto, Kyoto, Japan
Keyword: α-synuclein, GBA, Parkinson 's disease (PD), Gaucher's disease (GD)
[Objective] The GBA1 gene is responsible for Gaucher’s disease (GD). Recently, heterozygous mutations in GBA1 are known to be a strong risk factor for Parkinson's disease (PD) and dementia with Lewy bodies (DLB). In addition, non-neuronopathic GD has also been shown to develop PD or DLB at a high rate. However, there is still no disease modifying therapy for these disease. Previously, we have reported that the gba1 knock-out (KO) medaka display abnormal swimming movement, non-selective neuronal loss, and α-synuclein accumulation in the brains. These gba1 KO medaka are useful to investigate the research on GBA1-related disorder. [Methods] We have established a system in which candidate compounds are administered to gba1 KO medaka before the onset of motor symptoms in order to search for compounds that improve the GBA1-related disorder. As a primary screening, each candidates were administered to gba1 KO medaka from the 4 weeks post-fertilization, prior to the onset of motor symptoms, and the survival rate were analyzed. The effect of the compound that showed improved survival in the primary screening will be analyzed by histological and biochemical examination of the central nervous system.[Results] To date, primary screening has been performed on eight compounds and improved survival has been observed for two compounds. [Conclusions] We demonstrated the screening methods using gba1 KO medaka were useful to search for the efficacy of compounds. The two compounds found in primary screening may be a new disease modifying treatment for GBA1-related α-synucleinopathy. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-206
軸索変性誘導分子SARM1の阻害剤開発
Small molecule inhibitors targeting axon degeneration-inducer SARM1
*村田 等(1)、安藤 隆幸(2)、大磯 和真(1)、友信 奈保子(1)、山本 健一(1)、木下 理恵(1)、阪口 政清(1)
1. 岡山大学 学術研究院医歯薬学域 細胞生物学分野、2. 静岡県環境衛生科学研究所 医薬食品部
*Hitoshi Murata(1), Takayuki Ando(2), Kazuma Oiso(1), Nahoko Tomonobu(1), Ken-ichi Yamamoto(1), Rie Kinoshita(1), Masakiyo Sakaguchi(1)
1. Grad Sch Med Dent Pharma, Univ of Okayama, Okayama, Japan, 2. Dept of Drug Food Sci, SHIZUOKA INSTITUTE OF ENVIRONMENT AND HYGIENE
Keyword: SARM1, inhibitor, Axon degeneration, NAD+
Axonal degeneration is an early event that causes disability and disease progression in many neurodegenerative diseases. Sterile alpha and Toll/interleukin receptor motif-containing protein 1 (SARM1), which is highly expressed in neurons, has been attracting attention as an important molecule that induces axonal degeneration and neuronal cell death. Although SARM1 is usually inactive condition, when activated by stress stimulation, SARM1 metabolizes NAD+ and produces cyclic ADPR. Cyclic ADPR increases the intracellular Ca2+ concentration, and Ca2+-dependent calpain degrades axon components, resulting in axon degeneration and cell death. We have previously uncovered the mechanism by which JNK kinase phosphorylates and activates SARM1 under oxidative stress conditions. If the function of this active SARM1 can be inhibited, it may be possible to suppress axonal degeneration and contribute to the treatment of various neurodegenerative diseases. Therefore, we constructed a screening method for inhibitors using phosphorylated full-length SARM1, and searched for compounds that inhibit the NAD+-cleavage activity of active SARM1. From the screening of 1,920 small molecule compounds, three types (X, Y and Z) were found as compounds that inhibit the NAD+-cleavage activity of SARM1 by more than 50%. Addition of anticancer drugs to neurons as a model of chemotherapy-induced peripheral neuropathy (CIPN) caused neurite degradation and cell death, however compound X markedly suppressed these phenomena. Compound X also suppressed rotenone-induced neurite degradation and cell death that induce Parkinson's disease-like symptoms. Although administration of vincristine to mice as an in vivo model of CIPN showed hyperalgesia to mechanical and thermal stimuli and degradation of nerve fibers in the footpad, these symptoms were also improved by administration of compound X. These results suggest that inhibitors of active SARM1 can be applied to the treatment of neurodegenerative diseases. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-207
αシヌクレイン凝集体の鼻腔内単回投与により嗅球にLewy neurite様病理が生じる
Single intranasal administration of α-synuclein preformed fibrils induces Lewy neurites like pathology in the olfactory bulb.
*澤村 正典(1)、檜川 里衣(1)、大平 純一朗(1)、山門 穂高(1)、髙橋 良輔(1)
1. 京都大学付属病院脳神経内科
*Masanori Sawamura(1), Rie Hikawa(1), Junichiro Ohira(1), Hodaka Yamakado(1), Ryosuke Takahashi(1)
1. Kyoto University Hospital
Keyword: alpha synuclein, Parkinson's disease, intranasal administration
Background: Recent study suggests that the spread of α-synuclein (α-Syn) aggregates is involved in the progression of Parkinson's disease. The olfactory bulb (OB) is thought to be one of the initiation sites of α-Syn aggregates formation in the brain. It has been hypothesized that α-Syn aggregates possibly propagated from the nasal mucosa to the OB, however, there is no experimental evidence of α-Syn propagation from the olfactory mucosa to the OB in wild-type mice.
Objectives: To investigate whether single intranasal administration of mouse α-Syn preformed fibrils (PFFs) results in α-Syn propagation to the OB and central olfactory structures in wild-type mice.
Methods: Recombinant α-Syn was purified and incubated with agitation for a week to generate α-Syn fibrils. α-Syn fibrils were sonicated and administered into the unilateral nasal cavity of wild-type male mice, and mice were sacrificed at 1, 3, 6, and 12 months post-inoculation (mpi). The tissues were evaluated by immunostaining with anti-phosphorylated α-Syn (p-α-Syn), ubiquitin, and p62 antibodies.
Results: No p-α-Syn pathology was observed at 1 mpi, but Lewy neurite-like p-α-Syn aggregates first appeared at 3 mpi, and the number of aggregates gradually increased at 6 and 12 mpi. The p-α-Syn aggregates colocalized with ubiquitin or p62. P-α-Syn pathology was observed in the olfactory nerve and glomerulus in the olfactory bulb, but not in other central olfactory systems such as the anterior olfactory nucleus or the piriform cortex.
Conclusions: Single intranasal administration of α-Syn PFFs induced p-α-Syn pathology in the OB, suggesting that the α-Syn pathology in the OB can possibly originate from the nasal mucosa. In addition, this study suggests that inhalation of α-Syn PPFs may trigger α-Syn pathology in the central nerve system. We strongly recommend researchers to use masks and/or drafts to avoid the inhalation when using α-Syn PPFs. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-208
ヒトにおける神経特異的ELAVLタンパク質スプライシングアイソフォームの機能分担とALS病態との関連
Functional segregation of neuronal ELAVL protein splicing isoforms in human cells and their involvement in ALS pathophysiology.
*長谷川 実奈美(1)、恩田 亜沙子(1,2)、岡野 ジェイムス洋尚(1)
1. 東京慈恵会医科大学 総合医科学研究センター 再生医学研究部、2. 東京慈恵会医科大学 内科学講座 脳神経内科
*Minami Hasegawa-Ogawa(1), Asako Onda-Ohto(1,2), Hirotaka James Okano(1)
1. Division of Regenerative Medicine, Research Center for Medical Sciences, The Jikei University School of Medicine, 2. Department of Neurology, The Jikei University School of Medicine
Keyword: ALS, ELAVL, RNA-binding protein, splicing
Neuronal ELAVLs (nELAVLs: ELAVL2, 3 and 4 or HuB, C and D) are RNA-binding proteins expressed specifically in the nervous system. nELAVLs regulate splicing, mRNA stability and translation required for neuronal differentiation and axonal maintenance. Recently, several groups have shown the involvement of nELAVLs in pathogenesis of ALS, and the importance of nELAVLs in ALS studies has been increasingly recognized. It is known that nELAVLs have three RNA recognition motifs (RRM) and bind U-rich RNA elements to regulate splicing of target RNAs. Splicing isoforms of nELAVLs are caused by alternative splicing of exon 6 and part of exon 7, which corresponds to the hinge region between RRM2 and 3, and each isoform has a different splicing ability to regulate target RNAs. Interestingly, nELAVLs regulate the alternative splicing of other nELAVL family members, suggesting that their splicing isoforms are tightly regulated at the appropriate levels. Most studies of the nELAVLs splicing isoforms used rodent-derived tissues and cell lines because of the high amino acid sequence homology between human and rodent nELAVLs, however the ratio and role of the isoforms in human neuronal cells are still unknown. In this study, we analyzed the splicing patterns and functions of human nELAVLs isoforms in human iPS cell-derived neurons and SH-SY5Y human neuroblastoma line. Interestingly, the relative abundance of the splicing isoforms in human neurons is markedly different from those in the adult rodent brain. Furthermore, we showed that nELAVLs regulate the alternative splicing of TDP-43 exon 6 to produce short isoforms by pre-RNA binding via RRMs. Our findings suggest that nELAVLs might be involved in ALS pathophysiology by regulating the splicing patterns of TDP-43 isoforms. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-209
CRMP1のリン酸化は筋萎縮性側索硬化症モデルマウスの運動機能を調節する
Phosphorylation of CRMP1 modulates motor function of amyotrophic lateral sclerosis model mice
*浅野 徹也(1)、中村 治子(1)、川本 裕子(1)、多田 美紀子(1)、木村 弥生(2)、高野 洋志(3)、八尾 良司(3)、橋口 俊太(1)、高橋 慶太(1)、國井 美紗子(1)、田中 健一(1)、五嶋 良郎(4)、中村 史雄(5)、竹内 英之(1)、土井 宏(1)、田中 章景(1)
1. 横浜市立大学大学院医学研究科神経内科学・脳卒中医学、2. 横浜市立大学先端医科学研究センター、3. がん研究会・がん研究所・細胞生物部、4. 横浜市立大学大学院医学研究科分子薬理神経生物学、5. 東京女子医科大学医学部生化学
*Tetsuya Asano(1), Haruko Nakamura(1), Yuko Kawamoto(1), Mikiko Tada(1), Yayoi Kimura(2), Hiroshi Takano(3), Ryoji Yao(3), Shunta Hashiguchi(1), Keita Takahashi(1), Misako Kunii(1), Kenichi Tanaka(1), Yoshio Goshima(4), Fumio Nakamura(5), Hideyuki Takeuchi(1), Hiroshi Doi(1), Fumiaki Tanaka(1)
1. Dept Neurol & Stroke Med, Yokohama City Univ, Kanagawa, Japan, 2. Adv Med Res Ctr, Yokohama City Univ, Kanagawa, Japan, 3. Dept Cell Biol, Cancer Inst, Japanese Foundation for Cancer Res, Tokyo, Japan, 4. Dept Mol Pharmacol & Neurobiol, Yokohama City Univ, Kanagawa, Japan, 5. Dept Biochem, Tokyo Women's Med Univ, Tokyo, Japan
Keyword: ALS, CRMP1
[Objective] Collapsin response mediator proteins 1 (CRMP1) is an intracellular molecule that mediates Semaphorin-3A (Sema3A) signaling by phosphorylation at Ser522 (S522). Previous studies suggest that upregulation of Sema3A deteriorates motor function of amyotrophic lateral sclerosis (ALS). Using phosphoproteomic method, we identified that S522 phosphorylation of CRMP1 is enhanced in spinal cord of SOD1G93A mice, a mouse model of ALS. In this study, we aimed to reveal the role of CRMP1 in ALS pathogenesis by deletion (-/-) or abrogation of phosphorylation at S522 (ki/ki) of CRMP1 in SOD1G93A mice.[Methods]We newly generated CRMP1 knock-in mice in which the Ser residue at position 522 was replaced with Ala (CRMP1ki/ki). We assessed rotor rod performance and survival rate in CRMP1-/-/SOD1G93A and CRMP1ki/ki/SOD1G93A mice. We also evaluated pathological changes in spinal cord and neuromuscular junctions (NMJs) by immunohistochemistry. In addition, we also analyzed proteomic data of these mice to reveal the pathways underlying ALS pathogenesis.[Results]CRMP1ki/ki/SOD1G93A (n=20) and CRMP1-/-/SOD1G93A (n=20) mice showed longer and shorter latency to fall in rotarod test, respectively, compared to SOD1G93A (n=20) mice. Survival rate was prolonged in CRMP1ki/ki/SOD1G93Amice, but not in CRMP1-/-/SOD1G93A mice. Reflecting these phenotypic findings, the residual motor neurons and innervated NMJs were comparatively preserved in CRMP1ki/ki/SOD1G93A mice without affecting microglial and astroglial pathology. We also performed proteomic analysis using spinal cord and found that sirtuin signaling pathways were involved both in CRMP1-/-/SOD1G93A and CRMP1ki/ki/SOD1G93A mice. Among sirtuin signaling pathways, PGK1 fluctuated differently between these two lines of mice, and the spinal cord of CRMP1ki/ki/SOD1G93A was confirmed to show an increase in PGK1 expression by immunoblotting. [Conclusions]Here we demonstrated that inhibition of CRMP1 phosphorylation at S522 ameliorates the motor functions of ALS mice. Proteomic analysis suggested that CRMP1 might modulate ALS pathogenesis by altering sirtuin signaling pathways. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-210
線条体神経細胞の過活動抑制によるジスキネジアの治療
A therapeutic approach to reduce the hyperactivity of striatal neurons for controlling dyskinesias
*別宮 豪一(1)、パパ ステラ(2)、望月 秀樹(1)
1. 大阪大学大学院医学系研究科神経内科学、2. Yerkes霊長類研究所
*Goichi Beck(1), Stella M Papa(2), Hideki Mochizuki(1)
1. Dept Neurol, Osaka Univ Grad Sch Med, Suita, Japan, 2. YNPRC, Emory Univ, Atlanta, USA
Keyword: Parkinson's disease, L-dopa induced dyskinesia, striatum, medium spiny neuron
Background: Loss of nigrostriatal dopamine in Parkinson’s disease (PD) causes dysregulation of medium spiny neurons (MSNs) in the striatum. However, long-term dopamine replacement fails to efficiently modulate MSNs. Previous studies showed that recordings in animal models of advanced PD and patients have revealed significant hyperactivity of MSNs, and that hyperactive MSNs respond to dopaminergic stimulation with unstable firing changes, which are associated with the development of L-dopa induced dyskinesias (LIDs). Objective: The present study was aimed at examining whether decreasing the baseline MSN firing frequency would reduce the development of LIDs. Methods: To test acute pharmacological reduction of the MSN activity, a selective NMDAR antagonist (LY235959) or vehicle (aCSF) was infused into one side of the putamen of advanced parkinsonian non-human primate (NHPs, n=3). The antagonist was infused in the “off” state, and L-Dopa methyl ester plus benserazide was injected s.c. after the infusion. To test chronic reduction of the MSN activity, the inhibitory DREADDs (designer receptor exclusively activated by designer drugs) hM4Di was expressed in the striatum of hemiparkinsonian rats. rAAV-hSyn-hM4D(Gi)-mCherry or the control virus (rAAV-hSyn-GFP) was injected into the left striatum of rats with 6-hydroxydopamine lesions of the left nigrostriatal pathway (n=9). After 4 weeks, rats received daily clozapine N-oxide treatment to activate DREADDs and L-Dopa to induce abnormal involuntary movements (AIMs) for 2 weeks. The whole motor responses and LIDs/AIMs were assessed using standardized rating scales for NHP and rodents, respectively. Results: The NMDAR antagonist infusion in parkinsonian NHPs significantly reduced LID scores on the contralateral side without affecting the antiparkinsonian action of L-Dopa (motor disability scores of “on” state). Chronic activation of inhibitory DREADDs in rats significantly reduced AIMs scores when compared with those of the control group. Conclusions: These results indicate that strategies to reduce the hyperactivity of MSNs may inhibit the development of LIDs. Together data strongly support the reduction of MSN firing to improve L-Dopa responses and restore functionality in PD patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-211
内包脱髄モデルマウスによる再髄鞘化促進薬の新たな評価法
A novel evaluating method with the internal capsule demyelination mouse model for drugs promoting remyelination
*山崎 礼二(1)、長内 康幸(1)、幸喜 富(1)、篠原 良章(1)、大野 伸彦(1)
1. 自治医科大学
*Reiji Yamazaki(1), Yasuyuki Osanai(1), Tom Kouki(1), Yoshiaki Shinohara(1), Nobuhiko Ohno(1)
1. Jichi Medical University
Keyword: demyelination, oligodendrocyte, functional recovery, internal capsule
Multiple sclerosis (MS), a representative demyelinating disease in the central nervous system (CNS), is characterized by progressive remyelination failure, axonal injury and accumulation of motor deficits. Animal models of demyelination are frequently used to develop the potential therapy for MS. However, a tractable animal model to determine if remyelination promotes recovery of motor functions remained unavailable. Previously, we found that focal demyelination of internal capsule (IC) in mice by lysophosphatidylcholine (LPC) injection induced acute motor deficit and subsequent recovery through remyelination. However, it remained unknown whether IC demyelination mouse model can be used to evaluate functional recovery following the promotion of remyelination by drug treatment. In this study, we produced LPC-induced demyelination in IC of mouse brain, and treated those mice with clemastine, which is an anti-muscarinic drug and a candidate drug for MS. We demonstrated that clemastine treatment promoted the recovery of motor function and asymmetric motor disability. Moreover, density of mature oligodendrocyte was significantly increased, and axonal injury was reduced in the IC lesions. Furthermore, electron microscopy analysis of demyelinated IC showed that myelin thickness was recovered. Together, LPC-induced IC demyelination is a versatile animal model that allows the evaluation of both functional and histological recovery by promotion of remyelination in CNS lesions. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-212
多系統萎縮症におけるGCI形成メカニズム探索のための新規マウスモデル
The novel mice model to investigate the mechanisms of Glial cytoplasmic inclusions (GCIs) formation in multiple system atrophy (MSA)
*石本 智之(1)、山門 穂高(1)、大野 美樹(1)、梶 誠兒(1)、綾木 孝(1)、眞木 崇州(1)、松澤 秀一(1)、高橋 良輔(1)
1. 京都大学大学院医学研究科臨床神経学講座
*Tomoyuki Ishimoto(1), Hodaka Yamakado(1), Miki Oono(1), Seiji Kaji(1), Takashi Ayaki(1), Takakuni Maki(1), Shu-ichi Matsuzawa(1), Ryosuke Takahashi(1)
1. Department of Neurology Kyoto University Graduate School of Medicine
Keyword: Multiple system atrophy, alpha synuclein, oligodendrocyte, Glial cytoplasmic inclusions
[Objective] Pathological hallmarks of multiple system atrophy (MSA) and Parkinson's disease (PD) are alpha synuclein (αS) inclusions: αS aggregates are mainly observed in oligodendrocytes (OLGs) in MSA, whereas they are formed almost exclusively in neurons in PD. It has been an enigma how αS accumulates preferentially in OLGs in MSA, because OLGs express αS much less than neurons. Recent studies have reported that oligomeric αS is also observed in neurons even in the early stages of MSA, and that, in mice inoculated with synthetic αS fibrils, the accumulation of αS in OLGs is observed long after the accumulation of αS in neurons. These results suggest the possibility of neuronal origin of αS aggregates in MSA, but it is controversial because immunohistochemical evaluation of the formation of early αS aggregates in OLGs is technically difficult. The aim of this study is to investigate the mechanisms of GCIs formation in MSA by the sensitive detection of early αS aggregates in OLGs. [Method] [Result] We have generated transgenic mice that express human αS-GFP fusion proteins in OLGs under the control of the 2’, 3’-cyclic nucleotide 3’-phosphodiesterase (CNP) promoter (CNP αS-GFP Tg mice). In these mice the aggregation status of endogenous αS in OLGs can be monitored by GFP dot signals. 2-3 months old of CNP αS-GFP Tg mice were anesthetized with isoflurane and inoculated αS fibrils (N=18) or phosphate buffered saline (PBS) (N=3) into the dorsal striatum. The treated mice were sacrificed after 1 month, 4 months, and 12 months, respectively. CNP αS-GFP Tg mice inoculated with αS fibrils started to display GFP dots initially in the processes of OLGs, and the number and size of GFP dots were increased overtime in the centripetal manner. [Conclusion] CNP αS-GFP Tg mice are useful for detecting the spreading of αS aggregates in the intracellular space of oligodendrocytes and investigating the mechanisms of GCIs formation in MSA. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-213
前駆期パーキンソン病モデルマウスの自律神経障害及び貧血
Autonomic dysfunction and anemia in a mouse model of prodromal Parkinson's disease
*田口 智之(1)、生野 真嗣(1)、山門 穂高(1)、髙橋 良輔(1)
1. 京都大学大学院医学研究科臨床神経学
*Tomoyuki Taguchi(1), Masashi Ikuno(1), Hodaka Yamakado(1), Ryosuke Takahashi(1)
1. Department of Neurology Kyoto University Graduate School of Medicine
Keyword: Parkinson's disease, animal model, anemia, constipation
Purpose and method: Along with typical motor symptoms, patients with Parkinson's disease (PD) exhibits a variety of prodromal non-motor symptoms such as constipation, frequent urination, and orthostatic hypotension. Anemia is also reported in patients with PD and subjects with multiple prodromal symptoms of PD. We have previously reported a mouse model of Parkinson's disease (A53T-SNCA-BAC Tg mouse), which exhibits the olfactory dysfunction and RBD-like symptom. In this study, we investigated whether A53T-SNCA-BAC Tg mice have signs and symptoms observed in prodromal PD patients .Results: There is a decrease in the number of red blood cell and hemoglobin in A53T-SNCA-BAC Tg mice from 7 months of age (n=5). A53T-SNCA-BAC Tg mouse also showed an increased whole gut transit time by at least 9 months old (Wt n=6, Tg n=4). Pathological analysis showed that proteinase K (PK)-resistant phosphorylated α-synuclein(α-syn) was accumulated in the Auerbach's and Meissner's plexus of the intestinal tract(n=3). Moreover, accumulation of PK-resistant phosphorylated α-syn was also observed in the adrenal medulla, salivary gland, and lacrimal gland(n=3), which are under autonomic control.Conclusion: A53T-SNCA-BAC Tg mice showed anemia and decreased intestinal motility associated with phosphorylatedα-syn accumulation. The abnormal accumulation ofα-syn in glandular tissues such as salivary gland and lacrimal gland suggests the importance of pathologies of the autonomic nervous systems and is now attracting attention as peripheral biomarkers of PD. The relationship between anemia and systemicα-syn accumulation is now under investigation. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-214
OPTN-TIA1経路はストレス顆粒のクリアランスを促進し、ユビキチン化したTDP-43の凝集体形成を抑制する
Optineurin/TIA1 pathway reduces ubiquitinated TDP-43 aggregation by promoting stress granule clearance
*垣花 太一(1)、高橋 雅彦(1)、葛城 美徳(1)、山下 俊一(1)、神吉 智丈(1)、小野寺 理(2)、藤井 雅寛(1)
1. 新潟大学大学院医歯学総合研究科、2. 新潟大学脳研究所
*Taichi Kakihana(1), Masahiko Takahashi(1), Yoshinori Katsuragi(1), Shun-Ichi Yamashita(1), Tomotake Kanki(1), Osamu Onodera(2), Masahiro Fujii(1)
1. Niigata University, Graduate School of Medical and Dental Sciences, 2. Niigata University, Brain Research Institute
Keyword: ALS, Stress granules, TDP-43, Optineurin
Background
Amyotrophic lateral sclerosis (ALS) is a disease characterized by degeneration of motor neurons and is caused by aggregation of ubiquitinated TDP-43 protein (Ub-TDP-43) in the cytoplasm of motor neurons. Stress granules (SGs) are cytoplasmic protein aggregates containing TDP-43 and are induced by various stresses such as heat stress and oxidative stress. Previous studies have shown that delayed clearance of SGs promotes abnormal constitutive aggregation of Ub-TDP-43 in primary neurons of ALS patients and in neuronal cells with mutations in ALS-associated genes. Therefore, these abnormal SGs with delayed clearance are proposed to cause pathogenic aggregation of Ub-TDP-43 in ALS patients. Mutations in the optineurin (OPTN) and TIA1 genes have been found in familial ALS patients with the pathology of TDP-43 aggregation. In the present study, we analyzed the function of OPTN in Ub-TDP-43 aggregation and SG clearance.
Results
We found that OPTN decreases the amount of Ub-TDP-43 in non-neuronal cells and iPSC-derived primary neurons by promoting the clearance of SGs induced by heat shock stress. On the other hand, knockdown of OPTN or ALS-related OPTN mutants increased TIA1 at mRNA and protein levels, delayed SG clearance, and increased the amount of Ub-TDP-43, all of which were suppressed by TIA1 knockdown. Furthermore, overexpression of TIA1 delayed the clearance of SGs and increased the amount of Ub-TDP-43.
Conclusion
In this study, we found that depletion of OPTN or ALS-associated OPTN mutants induces aggregation of Ub-TDP-43 by increasing TIA1 level and delaying SG clearance. Like OPTN, ALS-associated TIA1 mutants have also been shown to induce aberrant SGs with delayed SG clearance. Our results suggest that the OPTN/TIA1 pathway suppresses pathological aggregation of Ub-TDP-43 in neurons by promoting SG clearance. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-215
パーキンソン病に伴う衝動的行動の神経基盤ー本邦での後方視的検討
Neural correlates of impulsive compulsive behaviours in Parkinson’s Disease - Japanese retrospective study
*木村 一皓(1,2)、梶山 裕太(1,3,4)、レバンカル ガジャナン(3)、小河 浩太郎(1,3)、天野 薫(2,5)、望月 秀樹(1,3)
1. 大阪大学医学部附属病院 神経内科・脳卒中科、2. 情報通信研究機構 未来ＩＣＴ研究所 脳情報通信融合研究センター、3. 大阪大学大学院 医学系研究科、4. 堺市立総合医療センター 脳神経内科、5. 東京大学大学院 情報理工学系研究科
*Ikko Kimura(1,2), Yuta Kajiyama(1,3,4), Gajanan Subhash Revankar(3), Kotarou Ogawa(1,3), Kaoru Amano(2,5), Hideki Mochizuki(1,3)
1. Department of Neurology, Osaka University Hospital, Suita, Japan, 2. Center for Information and Neural Networks (CiNet), Advanced ICT Research Institute, National Institute of Information and Communications Technology, Suita, Japan, 3. Graduate School of Medicine, Osaka University, Suita, Japan, 4. Department of Neurology, Sakai City Medical Centre, 5. Graduate School of Information Science and Technology, The University of Tokyo, Tokyo, Japan
Keyword: Parkinson's Disease, impulsive compulsive behaviours, functional connectivity, voxel based morphometry
Objectives: Parkinson’s Disease (PD) is a neuro-degenerative disease, characterised by decreased dopamine-production in the midbrain. Some PD patients present impulsive compulsive behaviours (ICB) such as pathological gambling and hypersexuality. ICB deteriorate patients’ and their caregivers’ quality of life, and nucleus accumbens (NAcc) was proposed to be involved in ICB, since NAcc is important in reward-based learning and its activation is affected by dopamine (Haber et al. 2010). However, the neural correlates of these behaviours in Japanese PD patients were unexplored. Here, we show that ICB were associated with functional connectivities (FC) of the right NAcc in PD patients.
Methods: 150 PD patients from our existing cohort study were investigated retrospectively. They underwent T1 weighted and resting state functional MRI scans. They were divided into two groups (ICB and non-ICB group) by the Japanese version of Questionnaire for Impulsive-Compulsive Disorders in Parkinson’s Disease (J-QUIP). In these groups, 74 patients matched for age, gender, and disease durations were analysed. To identify the functional characteristics of ICB, the FC of NAcc were compared between these two groups. Grey matter volumes (GMV) were also compared to identify structural correlates of ICB.
Results: Between ICB and non-ICB groups, we found no significant differences in levodopa equivalent daily dose or degree of motor symptoms, cognitive dysfunctions, depressions, and sleepiness. ICB group showed decreased FC between the bilateral orbito-frontal cortex (OFC) and the right NAcc, as well as increased FC between left middle occipital gyrus (MOG) and the right NAcc, compared with non-ICB group. We found no significant difference in the GMV between ICB and non-ICB groups.
Discussion: The OFC has projections to the NAcc and plays a crucial role in action inhibition. We speculate that the decreased FC between the right NAcc and the bilateral OFC in ICB group might reflect the disrupted regulation from the bilateral OFC to the right NAcc. By contrast, the increased FC between the right NAcc and the left MOG might reflect the disrupted visuo-spatial memory, which was reported in PD patients with ICB (Weintraub et al. 2017).
Conclusion: FC of the right NAcc were altered in ICB group. These connectivities can be useful biomarkers for detecting ICB in PD and might serve as new targets for treating these symptoms. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-216
遺伝子編集ヒトiPS細胞を用いた筋萎縮性側索硬化症の病態研究
Pathophysiological study of amyotrophic lateral sclerosis using gene edit human iPS cells
*恩田 亜沙子(1,2)、長谷川 実奈美(1)、松野 博優(1,2)、坊野 恵子(1,2)、平木 宏実(1,2)、鐘ヶ江 裕美(3)、井口 保之(2)、岡野 ジェイムス 洋尚(1)
1. 東京慈恵会医科大学 総合医科学研究センター 再生医学研究部、2. 東京慈恵会医科大学 内科学講座 脳神経内科、3. 東京慈恵会医科大学 総合医科学研究センター 基盤研究施設
*Asako Onda-Ohto(1,2), Minami Hasegawa-Ogawa(1), Hiromasa Matsuno(1,2), Keiko Bono(1,2), Hiromi Hiraki(1,2), Yumi Kanegae(3), Yasuyuki Iguchi(2), Hirotaka James Okano(1)
1. 1.Division of Regenerative Medicine, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo, Japan, 2. 2.Division of Neurology, The Jikei University School of Medicine, Tokyo, Japan, 3. 3.Core Research Facilities, Research Center for Medical Sciences, The Jikei University School of Medicine, Tokyo, Japan
Keyword: amyotrophic lateral sclerosis, induced pluripotent stem cells, CRISPR/Cas9, oxidative stress
【Background】Amyotrophic lateral sclerosis (ALS) is a disease with a poor clinical prognosis in which symptoms such as limb muscle weakness, dysphagia, and respiratory impairment follow a progressive course. The cause of ALS has not been clarified, and the fundamental treatment has not been found yet. About 10% of ALS cases are familial. TARDBP gene, which is one of the causative genes of familial ALS, encodes TAR DNA-binding protein 43kDa (TDP-43) protein, an RNA-binding protein mainly localized in the nucleus and involved in RNA transcription, splicing and translation. Interestingly, TDP-43-containing neuronal inclusions are formed in many patients with sporadic or familial ALS, regardless of the presence or absence of TARDBP mutations. From these findings, the molecular mechanism of TDP-43 may be the key to understand the pathology of ALS. Although motor neurons are selectively impaired in ALS, abnormal TDP-43 deposition in sensory neurons as well as motor neurons has been reported in autopsy cases of ALS, suggesting that sensory neurons may also be affected. In this study, we generate TDP-43 mutant induced pluripotent stem (iPS) cells from healthy human iPS cells and differentiate them into motor and sensory neurons, comparing the phenotype between healthy and disease groups. With this method, it will be possible to elucidate the molecular pathogenesis of ALS caused by TDP-43 mutations by comparing disease group cells with healthy group cells harboring identical DNA sequences except for TDP-43.
【Methods】We used CRISPR/Cas9 gene editing technology to establish TDP-43 mutant cell line from healthy human iPS cells with knock-in of A382T as well as WT cell line with knock-in of wild type sequences as a control because of FRT sequence insertion. After motor neurons and sensory neurons were differentiated from the control iPS cell line without gene editing, WT and A382T cell lines, the morphology, expression of specific-markers, mitochondrial morphology and axonal transport in the neurons were compared.
【Results】There was no difference in the phenotypes among the three groups after 2 months of culture. Oxidative stress induced apoptosis in group A382T motor neurons (P = 0.0124), however no significant difference was observed in sensory neurons, suggesting a specific vulnerability of motor neurons.
【Future Prospects】Our results indicated that A382T motor neurons seemed to be normal before loading oxidative stress which may reflect the fact that ALS takes many years to onset. Our iPS cell-based cellular ALS model enables to elucidate the molecular mechanisms underlining the vulnerability to stress in diseased motor neurons. In the future, this disease-model system will be able to contribute to implementation of High-Throughput Screening in drug discovery. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-217
運動ニューロンTDP-43病理はALS、PSP、CBDの共通した病態である
Motor neuron TDP-43 pathology links pathomechanisms of ALS, PSP, and CBD
*陸 雄一(1,2)、石垣 診祐(2)、藤岡 祐介(2)、長谷川 成人(3)、赤木 明生(1)、岩﨑 靖(1)、吉田 眞理(1)、勝野 雅央(2)、祖父江 元(1)
1. 愛知医科大学、2. 名古屋大学神経内科、3. 東京都医学総合研究所
*Yuichi Riku(1,2), Shinsuke Ishigaki(2), Yusuke Fujioka(2), Masato Hasegawa(3), Akio Akagi(1), Yasushi Iwasaki(1), Mari Yoshida(1), Masahisa Katsuno(2), Gen Sobue(1)
1. Aichi Med. Univ., 2. Dept. Neurol, Nagoya Univ., 3. Tokyo Metropolitan Institute of Medical Science
Keyword: TDP-43, motor neuron disease
[Objective] To assess TDP-43 pathology in the spinal cord motor neurons of tauopathies. [Methods] We examined 106 spinal cords from consecutively autopsied cases with progressive supranuclear palsy (PSP), corticobasal degeneration (CBD), globular glial tauopathy (GGT), Alzheimer's disease (AD), or Pick disease (PiD) and controls. [Results] We found mislocalization and cytoplasmic aggregation of TDP-43 in spinal cord motor neurons in 38% of PSP cases and 58% of CBD cases. The TDP-43-aggregates were molphologically similar to those of ALS and contained insoluble C-terminal fragments. The spinal cord TDP-43 pathology was often associated with TDP-43 pathology of the primary motor cortex. The severity of TDP-43 and 4-repeat tau pathologies in the cervical cord showed positive correlation, and these aggregates synergistically facilitated microgliosis. Spinal cord TDP-43 pathology did not develop in an age-dependent manner. We also assessed splicing factor proline/glutamine rich (SFPQ) expression, a recently-identified regulator of ALS/FTLD pathogenesis, in spinal cord motor neurons. Immunofluorescent and proximity-ligation assays revealed altered interactions between SFPQ and fused-in-sarcoma (FUS) in the neuronal nuclei of PSP, CBD, and ALS-TDP cases. Moreover, SFPQ expression was depleted in neurons containing TDP-43 or 4R-tau aggregates. [Conclusion] PSP and CBD may have properties of systematic motor neuron TDP-43 proteinopathy, suggesting mechanistic links with ALS-TDP. SFPQ dysfunction, arising from altered interaction with FUS, may be a candidate of the common pathway. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-218
疾患iPS細胞を用いたミトコンドリア病ドラッグスクリーニング
Drug screening for the treatment of mitochondrial dysfunction by using mitochondria disease patient-derived induced pluripotent stem cells
*本間 耕平(1)、坪田 一男(1)、岡野 栄之(2)、小澤 洋子(1,3)
1. 慶應義塾大学医学部眼科学教室、2. 慶應義塾大学医学部生理学教室、3. 聖路加国際大学眼科学教室
*Kohei Homma(1), Kazuo Tsubota(1), Hideyuki Okano(2), Yoko Ozawa(1,3)
1. Dept of Ophthal., Keio Univ Sch of Med, 2. Dept of Physiol., Keio Univ Sch of Med,, 3. Dept of Ophthal., St. Luke's Intl Univ
Keyword: Mitochondria, iPSC, MELAS, ROS
Mitochondria play an important role in the production of intracellular energy through oxidative phosphorylation, and several studies have suggested a relationship between mitochondrial dysfunction and aging-related diseases, including age-related macular degeneration (AMD). Mitochondrial disease induces apoptosis and dysfunction of retinal pigment epithelial cells (RPE), which support photoreceptor cells in the retina, and it is known that patients with mitochondrial diseases (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes, MELAS) often present with central retinal dystrophy. In this study, we utilized induced pluripotent stem cells derived from MELAS patients (MELAS-iPSCs) with the mitochondrial DNA (mtDNA) mutation A3243G to elucidate the mechanism of retinal diseases in MELAS patients. Live/Dead imaging of MELAS-iPSCs showed that inhibition of glucose uptake by 2-deoxyglucose (2-DG) caused cell death, which was partially rescued by treatment with taurine, a type of sulfated amino acid. Metabolomic analysis showed that the reduced (GSH)/oxidized (GSSG) glutathione ratio was decreased in MELAS iPSCs, and the levels of cysteine, a substrate of GSH, and oxidative stress markers were increased. Taurine normalized these changes, suggesting that MELAS iPSCs are affected by oxidative stress and that taurine partially mitigates the effects. MELAS-iPSCs were differentiated into retinal organoids and RPE (MELAS-RPE) by retinal 3D differentiation culture. The mtDNA mutation ratio of the differentiated MELAS-RPE was confirmed by quantitative PCR. Since AMD has been suggested to induce epithelial-mesenchymal transition (EMT) in RPE, we examined MELAS-RPE to find that EMT signaling was activated in MELAS-RPE, which was suppressed by taurine treatment. To further explore the effective drug targets, we screened the validated chemical compound library in 2-DG treated MELAS-iPSCs and found several drugs that could rescue the 2-DG-induced apoptosis in MELAS-iPSCs. We will also discuss this rescue mechanism and its potential therapeutic effect on the tissues of MELAS patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-219
アトラクチン欠損はミクログリアを活性化させドーパミンニューロン変性を引き起こす
Attractin deficiency promotes microglial activation resulting in dopaminergic neurodegeneration.
*江原 鮎香(1)、上田 秀一(1)、徳田 信子(1)
1. 獨協医科大学解剖学
*Ayuka Ehara(1), Shuichi Ueda(1), Nobuko Tokuda(1)
1. Dept Anat, Dokkyo Med Univ, Tochigi, Japan
Keyword: dopaminergic neuron, microglia, Attractin, neurodegeneration
Zitter rats, which are Attractin mutant rats, exhibit progressive dopaminergic neurodegeneration in the substantia nigra-striatal pathway. The mutant rats are also known to have increased activated microglia. This study aims to review the relationship between dopaminergic neurodegeneration and activated microglia in Zitter rats and clarify whether the deletion of endogenous Attractin directly activates microglia. In the substantia nigra of the Zitter rats, many swollen microglia formed clusters and surrounded the tyrosine hydroxylase-positive dopaminergic neurons. Microglial clusters were significantly abundant in the mutants from 2 weeks to 6 months of age compared to control SD rats. Next, the anti-inflammatory agent minocycline was chronically administered to Zitter rats for 6 months from 10 postnatal days. As a result, microglia changed to a normal thin shape, the number of microglial clusters decreased, and the overexpressed IL1b and iNOS were suppressed. Subsequently, chronic administration of minocycline protected dopaminergic neurons in the mutant against neurodegeneration. Finally, primary microglial cell cultures were performed to determine whether the deletion of endogenous Attractin directly causes microglial activation. There were no differences in morphology or IL1b expression levels between microglia from Zitter rats and control SD rats in the normal glial medium. However, LPS-induced stimulation of primary microglia led to overexpression of IL1b in microglia from Zitter rats compared with control rats. The level of IL1b overexpression increased exponentially in LPS dose-dependent manner. These studies indicate that Attractin-deficient microglia are highly sensitive to inflammatory stimuli and are prone to activation. These findings suggest that in Attractin deficiency, microglia activated during brain development remain active in adults, causing dopaminergic neurodegeneration. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-220
Zn結合型メタロチオネインによるALS関連タンパク質の凝集抑制
Zinc-binding metallothionein inhibits the SOD1 aggregation via binding of core regions for aggregate growth
*徳田 栄一(1)、桂馬 茉衣子(1)、池田 知亜子(1)、小菅 康弘(1)、小野 真一(2)
1. 日本大学薬学部、2. 阿伎留医療センター・神経内科
*Eiichi Tokuda(1), Maiko Keima(1), Chiako Ikeda(1), Yasuhiro Kosuge(1), Shin-ichi Ono(2)
1. Sch Pharm, Nihon Univ, Chiba, Japan, 2. Dep. Neurol., Akiru Med. Center
Keyword: amyotrophic lateral sclerosis, protein aggregation, metallothionein
Zn ions confer conformational stability to Superoxide disumutase-1 (SOD1). Loss of Zn ions compromises the stability of SOD1, leading to aggregation in motor neurons of amyotrophic lateral sclerosis (ALS). Thus, restoring Zn availability of SOD1 or milieu surrounding SOD1 is critical to inhibit the SOD1 aggregation. We have provided in vivo evidence for metallothionein (MT), a major cellular pool of Zn, in the SOD1 aggregation. Deletion of Mt gene in an ALS mouse model increases the amounts of SOD1 aggregates, whereas overexpression of MT decreases them. Moreover, MT is colocalized with SOD1 aggregates in the mouse model, suggesting their direct interaction. In this work, we performed in vitro studies to clarify whether MT could inhibit the SOD1 aggregation at molecular level. When Zn-deficient SOD1 is agitated in a test tube, SOD1 become cloudy or form insoluble aggregates. Thus, measuring the turbidity of the protein solution allows us to understand kinetics of SOD1 aggregation. In the absence of MT, Zn-deficient SOD1 was aggregated 20 min after the start of agitation. By contrast, Zn-binding MT (Zn-MT) significantly delayed the SOD1 aggregation, which was seen 180 min after agitation was initiated. This inhibitor effect was not observed in Cu-binding or metal-deficient MT, suggesting that Zn ions coordinated to MT is essential to inhibit the aggregation. We identified regions of SOD1 aggregate which Zn-MT is bound to. Zn-MT was bound to core regions of SOD1 aggregates, which is involved in the aggregate growth. In conclusion, Zn-MT inhibits the SOD1 aggregation at molecular level via binding of the aggregate core. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-221
パーキンユビキチン様ドメインの27番リジン残基でのユビキチン化は、パーキン活性化の新しいメカニズムを示唆している
Ubiquitination at the lysine 27 residue of the Parkin ubiquitin-like domain is suggestive of a new mechanism of Parkin activation
*劉 俊逸(1)、井下 強(1)、柴 佳保里(1)、服部 信孝(1)、今居 譲(1)
1. 順天堂大学医学研究科
*Junyi Liu(1), Tsuyoshi Inoshita(1), Kahori Shiba(1), Nobutaka Hattori(1), Yuzuru Imai(1)
1. Graduate School of Medicine, Juntendo University, Tokyo, Japan
Keyword: Parkin, PINK1, Mitochondria, Drosophila
The mitochondrial kinase PTEN‐induced kinase 1 (PINK1) and cytosolic ubiquitin ligase (E3) Parkin/PRKN are involved in mitochondrial quality control responses. PINK1 phosphorylates ubiquitin and the Parkin ubiquitin-like (Ubl) domain at serine 65 and promotes Parkin activation and translocation to damaged mitochondria. Upon Parkin activation, the Ubl domain is ubiquitinated at lysine (K) 27 and K48 residues. However, contribution of K27/K48 ubiquitination towards Parkin activity remains unclear. In this study, ubiquitination of K56 (corresponding to K27 in the human), K77 (K48 in the human), or both, was blocked by generating Drosophila Parkin (dParkin) mutants to examine the effects of Parkin Ubl domain ubiquitination on Parkin activation in Drosophila. The dParkin, in which K56 was replaced with arginine (dParkin K56R), rescued pupal lethality in flies by co-expression with PINK1, whereas dParkin K77R could not. The dParkin K56R exhibited reduced abilities of mitochondrial fragmentation and motility arrest, which are mediated by degrading Parkin E3 substrates Mitofusin and Miro, respectively. Pathogenic dParkin K56N, unlike dParkin K56R, destabilized the protein, suggesting that not only was dParkin K56N non-ubiquitin-modified at K56 but also the structure of the Ubl domain for activation was largely affected. Ubiquitin attached to K27 of the Ubl domain during PINK1-mediated Parkin activation was likely to be phosphorylated because human Parkin K27R weakened Parkin self-binding and activation in trans. Therefore, our findings suggest a new mechanism of Parkin activation, where an activation complex is formed through phospho-ubiquitin attachment on the K27 residue of the Parkin Ubl domain. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-222
ヒトおよびマウスの大脳皮質におけるパーキンソン病責任分子α-シヌクレインの発現プロファイル解析
Comparative analysis of differential expression of α-synuclein in the cerebral cortex between mouse and human brain
*田口 勝敏(1)、渡邊 義久(2)、辻村 敦(2)、田中 雅樹(1)
1. 京都府立医科大学 生体構造科学、2. 京都府立医科大学 基礎老化学
*Katsutoshi Taguchi(1), Yoshihisa Watanabe(2), Atsushi Tsujimura(2), Masaki Tanaka(1)
1. Dept. of Anatomy and Neurobiology, Kyoto Prefectual University of Medicine, 2. Dept. of Basic Geriatrics, Kyoto Prefectual University of Medicine
Keyword: α-Synuclein, Inhibitory synapse, Excitatory synapse, Cerebral cortex
α-Synuclein (αSyn), one of major components of Lewy bodies (LBs) and Lewy neurites (LNs), is physiologically expressed in presynapses and is involved in synaptic function. Abnormal intracellular aggregation of αSyn is observed as LBs and LNs in neurodegenerative disorders such as Parkinson’s disease or dementia with Lewy bodies. Accumulated evidence suggests that abundant expression of αSyn is one of the risk factors for neurodegeneration. Therefore, it is important to know the endogenous expression profiles of αSyn. Our previous studies demonstrated brain region-dependent differential expression patterns of αSyn using wild-type mice. Synaptic expression of αSyn is mostly accompanied by the expression of vesicular glutamate transporter-1 (vGluT-1), an excitatory presynaptic marker protein. In contrast, expression of αSyn in GABAergic inhibitory synapses is different among brain regions. For examples, in the cerebral cortex and hippocampus, αSyn is not observed in inhibitory synapses. On the other hand, αSyn is clearly expressed in inhibitory synapses of the globus pallidus and substantia nigra pars reticulata. These results indicate that expression of αSyn is regulated in cell-type or brain-region dependent manner. Here, we studied expression profiles of αSyn in human brain. In the cerebral cortex, αSyn was mainly expressed in the excitatory synapses expressing vGluT-1, as happens in mouse brain. Interestingly, there were also a few αSyn-positive synapses co-expressing inhibitory marker proteins such as glutamic acid decarboxylase, vesicular GABA transporter, and cholecystokinin in human brain. These results suggested that expression profile of αSyn in inhibitory synapses was different between rodents and primates. Further studies of the differential expression of αSyn will provide new insights for understanding physiological function of αSyn. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-223
Phenotypic heterogeneity in the cerebellum in two lines of Inpp4a mutant mice
*Dang Minh Tran(1), Nozomu Yoshioka(1), Junko Sasaki (2), Takehiko Sasaki (2), Hirohide Takebayashi (1)
1. Grad Sch Med & Den, Univ of Niigata, Japan, 2. Med Research Institute, Tokyo Med & Den Univ, Japan
Keyword: Inpp4a, Phosphoinositides, Cerebellum
Phosphoinositides are a group of lipids that are present in the inner leaflets of plasma membranes and intracellular organelles. They regulate various intracellular signaling and membrane trafficking. Their actions play an important role in cell growth, metabolism, and cell death. Phosphoinositide metabolism are showed to be contributed to diseases such as neurological disease, cancer, diabetes, myopathy and inflammation. Our study focuses on the role of inositol polyphosphate-4-phosphatase type I (Inpp4a), one of phosphatases involving in phosphoinositide turnover. Recent studies have indicated that Inpp4a disfunction is linked to neurodegenerative disorders. For example, Inpp4a mutant Weeble mice, which carries a spontaneous loss-of-function mutation in Inpp4a gene (Inpp4awb/wb), show severe motor instability, cerebellar degeneration and ataxia. Gene targeting of exon encoding C-terminal phosphatase domain of Inpp4a (Inpp4aΔ23/Δ23) also leads to neurodegeneration in the cerebellum and striatum. However, the other Inpp4a KO mouse line harboring mutation in the N-terminus (first and second coding exons, Inpp4aΔ1,2/Δ1,2) exhibited neurodegeneration in the striatum, however, attenuated phenotype in the cerebellum. Here, we compare phenotypes between the two Inpp4a mutant lines and discuss about the underlying mechanism. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-224
腸管脂質センサーを起点とした腸脳相関によるPGD2－ミクログリア神経炎症サイクルの抑制
Brain-gut interaction originate from intestinal lipid sensor suppresses the PGD2-microglia-neuroinflammation cycle
*吉川 圭介(1)、岩佐 健介(1)、山本 梓司(1)、春田 力(1)、丸山 敬(1)
1. 埼玉医科大学医学部薬理学教室
*Keisuke Yoshikawa(1), Kensuke Iwasa(1), Shinji Yamamoto(1), Chikara Haruta(1), Kei Maruyama(1)
1. Department of Pharmacology, Faculty of Medicine, Saitama Medical University
Keyword: Prostaglandin, Microglia, GPR120, Neuroinflammation
Neuroinflammation is a key pathological component of neurodegenerative disease and is characterized by microglial activation and the secretion of proinflammatory mediators. We previously reported that a surge in prostaglandin D2 (PGD2) production and PGD2-induced microglial activation could provoke neuroinflammation. We also reported that a lipid sensor GPR120, which is expressed in intestine, could be activated by polyunsaturated fatty acids (PUFA), thereby mediating secretion of glucagon-like peptide-1 (GLP-1). To reveal the relationship between PGD2-microglia-provoked neuroinflammation and intestinal PUFA/GPR120 signaling, we investigated neuroinflammation and neuronal function in GPR120 knockout (KO) mice. In the current study, we discovered notable neuroinflammation (increased PGD2 production and microglial activation) and neurodegeneration (declines in neurogenesis, hippocampal volume, and cognitive function) in GPR120 KO mice. Our in vitro primary microglial cell cultures revealed that H-PGDS expression increased PGD2 production and microglial Iba-1 expression, which could be blocked by DP1- and DP2-antagonists. These data suggesting that microglia activated by PGD2 in an autocrine manner caused PGD2-microglia neuroinflammation cycle. We also found that inhibition of PGD2 production and potentiation of GLP-1 bioactivity reduced PGD2-microglia neuroinflammation cycle and further neurodegeneration in GPR120 KO mice. These observations raise the possibility that intestinal GLP-1 secretion, stimulated by intestinal GPR120, may remotely contributed to suppress PGD2-microglia neuroinflammation cycle in the hippocampus. These findings may suggest that insensitivity to dietary PUFA by dysfunction of GPR120 would raise the risk of hippocampal dysfunction. These observations may reveal the presence of a novel gut-brain interaction, in that the signaling of dietary PUFA is sensed by GPR120, converted into incretin bioactivity, and contributes to hippocampal homeostasis via suppression of PGD2-microglia neuroinflammation cycle. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-225
脳におけるリン脂質フリッパーゼの機能解明
To clarify the function of phospholipid flippase in the brain
*梅村 悠太(1)、大嶋 智葉(1)、中島 鼓美(1)、松田 拓与(1)、川瀬 宗之(1)、徳永 柊(1)、大石 久史(2)、築地 仁美(1)、河野 孝夫(1)、服部 光治(1)
1. 名古屋市立大学大学院薬学研究科 病態生化学分野、2. 名古屋市立大学大学院医学研究科 病態モデル医学分野
*Yuta Umemura(1), Tomoha Ohshima(1), Tsuzumi Nakajima(1), Takuto Matsuda(1), Muneyuki Kawase(1), Shu Tokunaga(1), Hisashi Oishi(2), Hitomi Tsuiji(1), Takao Kohno(1), Mitsuharu Hattori(1)
1. Department of Biomedical Science, Graduate School of Pharmaceutical Sciences, Nagoya City University, Aichi, Japan, 2. Department of Comparative and Experimental Medicine, Graduate School of Medical Sciences, Nagoya City University, Aichi, Japan
Keyword: Flippase, Glial cells, CAMRQ
There is an asymmetry in the phospholipid bilayers in biological membranes, with phosphatidylserine (PS) and phosphatidylethanolamine (PE) being found mainly on the inner leaflet, and phosphatidylcholine and sphingomyelin mainly on the outer leaflet. Phospholipid flippases, which belong to the P4-ATPases, contribute to the maintenance of the phospholipid asymmetry by translocating PS and PE from the extracellular side to the cytoplasmic one. Excluding PS from the outer leaflet is particularly important to prevent excessive phagocytosis. ATP8A2, a member of P4-ATPases, is expressed mainly in the nervous system, and mutations in ATP8A2 in humans are responsible for cerebellar ataxia, mental retardation, and disequilibrium syndrome (CAMRQ). ATP8A1, which is homologous to ATP8A2, is abundantly expressed in the brain and is reported to play a role in memory formation. These findings indicate that flippases are important for the normal development and function of the brain, but it is unknown in which cell types of the brain flippases function and how flippases work in those cell types to maintain normal brain function. We aim to elucidate the cellular functions of the flippases in the brain.
We generated ATP8A1-deficient (ATP8A1 KO) and ATP8A2-deficient (ATP8A2 KO) mice respectively, using the CRISPR/Cas9 system. We then crossed them to generate ATP8A1/ATP8A2 double-deficient (DKO) mice. We found in the cerebellum of ATP8A2 KO mice, that neurodegeneration occurred and that the number of activated microglia and astrocytes increased. These abnormalities were more significant in the cerebellum of DKO mice than in ATP8A2 KO mice. In the cerebral cortex and hippocampus of DKO mice, but neither of the single KO mice, neurodegeneration and the activated astrocytes were observed. Interestingly, the degree of glial cell activation was more prominent than that of neurodegeneration in many regions of the brain. Therefore, it was suggested that ATP8A1 and ATP8A2 work redundantly in many areas of the brain and that the primary role of the flippases was not to inhibit neurodegeneration but to suppress unnecessary activation of glial cells by inhibiting PS exposure. As activated glial cells are known to contribute to the progression of neurodegeneration, inhibition of the PS exposure or blocking PS receptors or the glial cells may ameliorate ATP8A2-related disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-226
マウス神経細胞内におけるα-シヌクレイン凝集体局在部位の経時変化解析
Temporal changes of subcellular locations of alpha-synuclein aggregates in mice
*亀田 浩司(1)、岡本 慎一郎(1)、小池 正人(1)
1. 順天堂大学大学院医学研究科
*Hiroshi Kameda(1), Shinichiro Okamoto(1), Masato Koike(1)
1. Grad Sch Med, Juntendo University, Tokyo, Japan
Keyword: Neurodegenerative disease, propagation, VGLUT1
Alpha-synuclein (α-syn) is a highly conserved 140-amino acid presynaptic protein which is thought to be involved in neuronal plasticity. This protein is also known to be the pathogenic protein in Parkinson’s disease, dementia with Lewy bodies and multiple system atrophy, which are collectively called synucleinopathy. Aggregated α-syn propagates mainly retrogradely and transsynaptically, and causes depression of cellular function. In this study, we injected sonicated α-syn preformed fibrils (PFFs) into the CPu of mice and detected the aggregated α-syn using immunohistochemistry. The PFFs are also believed to be transported retrogradely and transsynaptically and initiate aggregation of intrinsic α-syn, resulting in motor deficits in mice. One month after the PFFs injections, small punctiform signals were mainly observed in the CPu. Line-shaped and spherical signals were sparsely detected in the neocortex. Two months after the injections, the number of the small punctiform signals in the CPu and the line-shaped and spherical signals in the neocortex were increased. In addition, many line-shaped signals were detected in the corpus callosum and external capsule. On the other hand, the numbers of the small punctiform signals in the CPu and the line-shaped signals in the corpus callosum and external capsule were decreased four months after the injections. Instead, the line-shaped and spherical signals were increased in the CPu. Some small punctiform signals were immunopositive for axon markers such as vesicular glutamate transporter 1, whereas the spherical signals hardly colocalized. On the other hand, the spherical signals often existed around the NeuN-positive cell bodies. These results indicate that α-syn migrates within the neurons retrogradely and propagates transsynaptically. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-227
筋萎縮性側索硬化症モデルマウスにおける免疫疲弊分子の発現解析
Expression patterns of immune exhaustion molecules in the mouse model of amyotrophic lateral sclerosis
*森﨑 祐太(1)、青野 らん(1)、大島 基希(1)、岡部 亜美(1)、寺川 真代(1)、山中 宏二(2)、三澤 日出巳(1)
1. 慶應義塾大学薬学部 薬理学講座、2. 名古屋大学環境医学研究所 病態神経科学分野
*Yuta Morisaki(1), Ran Aono(1), Motoki Ohshima(1), Ami Okabe(1), Mayo Teragawa(1), Koji Yamanaka(2), Hidemi Misawa(1)
1. Div. Pharmacol., Fac. Pharmacy, Keio Univ., 2. Dept. Neurosci. & Pathobiol., Res. Inst. Environ. Med., Nagoya Univ.
Keyword: ALS, immune checkpoint, immune exhaustion, neuroinflammation
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by the progressive death of motor neurons. Neuroinflammation by glial cells and other immune cells infiltrating in the central nervous system has been suggested as a mechanism of motor neurodegeneration. Under chronic inflammatory conditions, immune cells express immune checkpoint molecules (immune exhaustion molecules, e.g. PD-1 ,LAG-3 and TIM-3), which regulate the reactivity of immune cells by suppressing excessive activation and autoimmune responses. Therefore, abnormalities in the expression and function of immune exhaustion molecules induce the disruption of immune tolerance and may be involved in the pathogenesis of neurodegenerative diseases.
In this study, we analyzed the expression patterns of immune exhaustion molecules in the mouse model of ALS (SOD1G93A mouse). In peripheral immune tissues such as spleen and lymph nodes, the mRNA expression level of immune exhaustion molecules in SOD1G93A mice was comparable to wild-type mice. In the spinal cord of SOD1G93A mice, however, we found that its expression level was increased compared with wild-type mice. We performed immunohistochemical analysis in the spinal cord of SOD1G93A mice, by multi-staining for immune exhaustion molecules and glial cell markers, and identified PD-1 and LAG-3 were expressed in microglia as the disease progressed. The increased expression of these molecules in microglia was not observed in other inflammation-induced gliosis models. Furthermore, both membranous and soluble forms of immune exhaustion molecules were upregulated in the spinal cord of SOD1G93A mice at late disease stage. These results suggest that the expression of immune exhaustion molecules in the spinal cord was increased in accordance with the disease progression, which may be involved in the pathogenesis of ALS by affecting immune tolerance. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-228
筋萎縮性側索硬化症におけるDEAD-boxヘリカーゼDDX5及びDDX17の病理学的解析
Pathological analysis of DEAD-box RNA helicases, DDX5 and DDX17 in amyotrophic lateral sclerosis.
*池田 真悟(1)、土井 宏(1)、多田 美紀子(1)、橋口 俊太(1)、高橋 慶太(1)、國井 美紗子(1)、田中 健一(1)、児矢野 繁(2)、竹内 英之(1)、田中 章景(1)
1. 横浜市立大学神経内科学・脳卒中医学、2. 横浜南共済病院神経内科
*Shingo Ikeda(1), Hiroshi Doi(1), Mikiko Tada(1), Shunta Hashiguchi(1), Keita Tkahashi(1), Misako Kunii(1), Kenichi Tanaka(1), Shigeru Koyano(2), Hideyuki Takeuchi(1), Fumiaki Tanaka(1)
1. Department of Neurology and Stroke Medicine, Yokohama City University Graduate School of Medicine, 2. Department of Neurology, Yokohama Minami Kyousai Hospital
Keyword: ALS, DDX5, DDX17
Accumulation of insoluble proteins in neurons and glial cells are common and important pathological features in neurodegenerative diseases. In this study, we investigated the pathological involvement of DEAD box RNA helicases, DDX5 and DDX17 in sporadic amyotrophic lateral sclerosis (SALS), which we previously identified as nuclear polyglutamine aggregate-interacting proteins in Huntington disease cell model. We immunohistochemically analyzed the subcellular distribution of DDX5 and DDX17 in the spinal cords of 30 SALS and 15 control autopsied cases. We also checked whether the intracellular dynamics of DDX5 and DDX17 are affected by mutant TDP-43 or cellular stress in cultured cells. We found that DDX5 formed neuronal cytoplasmic inclusions (NCIs) which were co-localized with TDP-43 in 22% (n=7/30) of SALS cases. In contrast, DDX17 formed small granular structures co-localizing with endoplasmic reticulum markers, but not with TDP-43 in all SALS cases. In HEK293T cells overexpressing mutant TDP-43, DDX5 co-localized with cytoplasmic TDP-43 aggregates, while the distribution of DDX17 was not affected. The amounts of DDX5 and DDX17 in the insoluble fractions were increased by overexpression of mutant TDP-43 compared to wild-type TDP-43. Overexpression of DDX5 caused aggregate formation in cytoplasm, which co-localized with TDP-43, while overexpression of DDX17 did not alter the distribution of TDP-43. Interestingly, the endoplasmic reticulum-like granular structure formation of DDX17 in SALS was reproduced in HEK293T cells by the administration of tunicamycin, which was known to cause the endoplasmic reticulum stress. Our results suggest that the RNA helicase DDX5 and DDX17 are differentially implicated in SALS pathology. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-229
常染色体優性遺伝性脊髄小脳変性症家系におけるPRKCG遺伝子の新規変異の同定
Novel PRKCG mutations in Japanese autosomal dominant spinocerebellar ataxia
*多田 有似(1)、久米 広大(1)、野口 颯真(2)、大澤 亮介(1)、酒井 規雄(2)、川上 秀史(1)
1. 広島大学原爆放射線医科学研究所、2. 広島大学大学院医系科学研究科
*Yui Tada(1), Kodai Kume(1), Soma Noguchi(2), Ryosuke Ohsawa(1), Norio Sakai(2), Hideshi Kawakami(1)
1. RIRBM, Hiroshima Univ, Hiroshimaa, Japan, 2. Grad Sch Biomed & Health Sci, Hiroshima Univ, Hiroshimaa, Japan
Keyword: Spinocerebellar ataxia, PRKCG
Spinocerebellar ataxia (SCA) is a neurodegenerative disease with spinal or cerebellar ataxia as its main symptom. A number of causative genes have been reported, and the disease types are distinguished by the genes. The most frequent genetic mutation of casative gene is an abnormal expansion of a repeat sequence in the coding region. In addition, point mutations in some causative genes have been also identified to develop SCA. However, because SCAs caused by missense mutations are less fluent, the details of their onset mechanism are still unknown. In order to understand the whole picture of SCA, it is significant to identify and investigate these rare mutations of the causative genes.
We performed exome analysis on autosomal dominant hereditary SCA families who had no abnormal repeat expansions of known causative genes. As a result, we identified two novel missense mutations in protein kinase C gamma (PRKCG) gene in two Japanese families: c.171G>C, p.W57C and c.400T>C, p.C134R. PRKCG gene encodes γPKC and reported as causative gene of SCA14. Both W57C and C137R mutations are present in the regulatory region of γPKC protein. In the family with the C137R mutation, only the affected individuals had the mutation, and the non-affected individual did not.
When these two mutant PRKCGs were expressed in HeLa cells and their localization of the protein was observed, both two mutant γPKC proteins aggregated in cytoplasm. Furthermore, we evaluated the solubility of these mutant proteins. As a result, the solubility of C137R γPKC protein was decreased compared to that of wild γPKC. On the other hand, there was no significant change in the solubility of W57C γPKC. This could explain why the symptoms of W57C patients were modelate. These results strongly suggest that these two mutations are new missense mutations that cause SCA14. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-230
Microtubule affinity-regulating kinases contribute differently to tau-induced neurodegeneration
*Grigorii Sultanakhmetov(1), Akiko Asada(1), Kanae Ando(1)
1. Graduate School of Science, Tokyo Metropolitan University, Tokyo, Japan
Keyword: Tauopathy, Drosophila
Abnormal phosphorylation of tau protein leads to its unbinding from microtubule, aggregation, and accumulation. That tau misbehavior directs to several neurodegenerative diseases called tauopathies. The phosphorylation of tau protein at sites Ser262 and Ser356 in microtubule-binding repeats by Par1/microtubule affinity-regulating kinase (MARK) could be an initial step of tau abnormality. Par-1/MARK are highly conserved AMPK-related kinases that form four members in mammals (MARK1-4). Although the association of tau toxicity with its phosphorylation by MARKs was established, it is not clear whether and how each member of the MARK family affects tau phosphorylation and toxicity in vivo. To address this question, we utilized a Drosophila model of tauopathy, in which human tau expression causes neurodegeneration. We established transgenic Drosophila carrying human MARK1-4 cDNA under the control of UAS promoter and co-expressed each MARK with human tau in fly eyes using a pan-retinal driver GMR-GAL4. Western blot analyses revealed that co-expression of MARK1, 2, and 4 increased phosphorylated tau both at Ser262 and Ser356, while MARK3 promoted phosphorylation only at Ser262. MARK4 caused accumulation of tau, but MARK1-3 did not affect tau protein levels. MARK4 does not cause accumulation of tau carrying alanine substitutions at Ser262/356 with alanine (S2A), suggesting that MARK4 causes accumulation of phosphorylated tau. MARK4 increased tau mRNA levels two-fold, while MARK4 also increased mRNA levels of S2A tau: thus, elevated mRNA levels do not correlate with tau protein accumulation. MARK1-4 all promoted neurodegeneration, while they did not do so equally: co-expression of human tau with MARK2 or MARK4 showed more severe neurodegeneration than that with MARK1 and MARK3. In addition, we found that the MARK2 expression was higher than MARK4, suggesting that MARK4 has a more substantial effect on neurodegeneration than MARK2. Our results revealed that mechanisms by which each member of the human MARKs affects tau-induced neurodegeneration via different mechanisms and suggest that MARK4 enhances the toxic effect of tau the most prominently. Funding sources: the Japan Foundation for Aging and Health Research Award and JSPS KAKENHI grant #19K21593 to KA. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-231
正常眼圧縁内障モデルマウスにおける視神経伸長を促す薬物の検討
Investigation of drug effects on optic nerve growth in a mouse model of normal tension glaucoma
*渋江 省吾(1)、東田 千尋(1)
1. 富山大学和漢医薬学総合研究所
*Shogo Shibue(1), Chihiro Tohda(1)
1. Section of Neuromedical Science, Institute of Natural Medicine, University of Toyama
Keyword: glaucoma
Glaucoma is a major cause of irreversible blindness worldwide characterized by degeneration and loss of retinal ganglion cells (RGC). Glaucomatous optic nerve neuropathy is caused by progressive atrophy of the vision system. All current drug treatments are focused on lowering intraocular pressure, which don’t accomplish vision recovery. Our laboratory previously found axonal regeneration activity of Drug A (name is closed due to the patent). Therefore, this study aimed to investigate effects of Drug A on optic nerve growth in an optic nerve crush model. As a normal tension glaucoma model, optic nerves were crushed in mice. Our preliminary study suggested that intravitreal injection of Drug A increased optic nerve density in the optic nerve crush model. This study aims to develop orally treatable medicines. Therefore, we investigated brain penetration of Drug A after p.o. administration. One, six and twenty-four hours after p.o. administration of Drug A to mice, the retina, the optic nerve and whole brain were isolated after complete perfusion to serve for LC-MS analysis. As a result, Drug A in those tissues was detected in those tissues. Immediately after optic nerve crushing, Drug A or vehicle solution was administered orally for 3 weeks. Five days before sacrificing, anterograde tracer, Alexa Fluor-conjugated cholera toxin B (CTB) was intravitreally injected. Intraocular pressure measurements and a behavioral vision test were conducted on the day of sacrificing. After that, the retina, optic nerves and brain were dissected and served for histochemical evaluation. The number of retinal ganglion cells and the density of optic nerve were quantified. Optic nerve termination to the lateral geniculate nucleus and superior colliculus are now under investigation. Drug A treatment to primary cultured retinal ganglion cells (RGC) for 4 days significantly enhanced axon length. This study investigates the effects of Drug A on optic nerve termination and visual function. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-232
Serum neurofilament light chain associated with outcome in Guillain-Barré syndrome across regions and disease spectrum
*Hoehn Benjamin(1)、Islam Zhahirul (2)、Sheikh Kazim(3)、Harbo Thomas (4)、Jahan Israt(2)、Hayat Shoma(2)、Kroon Henk-André(1)、Cahir-McFarland Ellen(1)、Yednock Ted(1)、Keswani Sanjay(1)、Deen Mohammad Quazi(5)
*Benjamin Hoehn(1), Zhahirul Islam(2), Kazim A. Sheikh(3), Thomas Harbo(4), Israt Jahan(2), Shoma Hayat(2), Henk-André Kroon(1), Ellen Cahir-McFarland(1), Ted Yednock(1), Sanjay Keswani(1), Quazi Deen Mohammad(5)
1. Annexon Biosciences, Inc., Brisbane, CA, USA, 2. Laboratory of Gut-Brain Signaling, Laboratory Sciences and Services Division, icddr,b, Dhaka, Bangladesh, 3. Department of Neurology, McGovern Medical School, Houston, TX, USA, 4. Department of Neurology, Aarhus University Hospital, Aarhus, Denmark, 5. National Institute of Neurosciences and Hospital, Dhaka, Bangladesh
Keyword: Guillain-Barré syndrome, neurofilament-light, prognostic biomarker, complement
Introduction: Guillain-Barré syndrome (GBS) is an acute classical complement mediated peripheral nerve disease with a diverse regional presentation. Neurofilament light chain (NfL), a biomarker of neuronal damage, is elevated in GBS. The objective of this study was to evaluate serum NfL (sNfL) as a prognostic biomarker in patients with GBS from different geographical areas. Methods: Serum samples were collected prospectively from 35 healthy controls and 97 GBS patients, 82 from Bangladesh and 15 from Europe and North America (EU/NA). Patients >=17 years of age, with onset of weakness <=2 weeks and a GBS disability score (GBS-DS) of <=3 were included. Serum NfL was assessed using single-molecule array and its association with GBS-DS was evaluated by Spearman’s coefficient, unadjusted for known prognostic factors and irrespective of treatment administered. Results: All patients had elevated sNfL, which correlated with GBS-DS (r=0.5395, p<0.0001), peaked 12.5 days after admission, declined by week 4, and normalized by week 26. Baseline mean sNfL was 932.1±1274.2 pg/mL (1-1200x above controls). As a monophasic disease, peak sNfL (psNfL) reflected GBS severity most appropriately (r=0.136, p=<0.001) and provided more informative correlations than baseline sNfL with age (r=-0.371, p<0.001), ability to improve in GBS-DS by weeks 4 (r=0.480, p<0.0001) or 8 (r=0.351, p<0.001), and GBS-DS at month 6 (r=0.55, p<0.0001). Higher psNfL (1986.0 vs. 1050.0 pg/mL; p=0.07) was seen in patients with acute motor axonal neuropathy (AMAN) vs. acute inflammatory demyelinating polyneuropathy (AIDP). At 6 months, 45.6% of patients could run (GBS-DS <=1), which was associated with a median psNfL of 362.2 vs. 1417.0 pg/mL (p=0.0025) for those patients unable to run (GBS-DS >=2). In Bangladesh, 11 out 32 patients (34.4%) could run compared to 10 out of 14 patients (71.4%) in Europe/North America (EU/NA). Consistent with a psNfLBangladesh : psNfLEU/NA ratio of 5.95, median psNfL was 766.0 vs. 2298.0 pg/mL (p=0.088) in Bangladesh and 182.2 vs. 455.1 pg/mL (p=0.142) in EU/NA comparing GBS-DS <=1 vs. GBS-DS >=2. Median sNfL levels corresponded with the 40th percentile rank in the sNfL distribution in each population. In patients able to run the AMAN:AIDP ratio was similar across the 2 groups. Conclusions: Elevated sNfL mirrored disease course and correlated with severity, axonal variants, and functional outcome in a heterogeneous population from different regions. Integration of serum NfL in a universal GBS disease model is warranted. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-234
Emodinは変異ＡＲを減らして球脊髄性筋萎縮症モデルの表現型を改善する
Emodin alleviates SBMA phenotype via down regulation of androgen receptor
*足立 弘明(1)、強 強(1)、豊田 知子(1)、勝野 雅央(2)、祖父江 元(2)
1. 産業医科大学、2. 名古屋大学大学院医学系研究科
*Hiroaki Adachi(1), Qiang Qiang(1), Tomoko Toyota(1), Masahisa Katsuno(2), Gen Sobue(2)
1. Sch Med, Univ of Occupational and Environmental Health, Kitakyushu, Japan, 2. Nagoya University Graduate School of Medicine
Keyword: Spinal and bulbar muscular atrophy, Androgen receptor, emodin, heat shock protein 90
Spinal and bulbar muscular atrophy (SBMA) is an x-linked hereditary neuromuscular disorder caused by the expansion of CAG repeats in the first exon of androgen receptor (AR) gene. The expanded CAG repeats encode a long polyglutamine tract in the N-terminal transactivation domain of AR protein. SBMA characterized clinically by limb weakness, fasciculation, and muscle atrophy. The pathologic feature of SBMA is selectively loss of motor neurons in the anterior horn of spinal cord and the bulbar region of brain stem, diffusely accumulated inclusions of mutant AR in the nuclear of the residual motor neurons. Currently there is no effective treatment available for this disease. Because the expanded CAG repeats mutation in AR gene is likely linked to toxic gain of function, decreasing the amounts of mutant AR protein is becoming an optional therapy avenue for the SBMA treatment. We examined the effects of emodin in cultured cell models of SBMA. Cells were transfected using Lipofectamine 2000 with plasmids encoding mutant AR containing normal (24 CAGs) or expanded (97 or 112 CAGs) polyQ repeats. 4 or 40 mg/kg of emodin was administered intraperitoneally every day. Control mice received DMSO. We demonstrated that emodin, a natural compound extracted from Chinese herbs, could decrease the association of mutant AR with heat shock protein 90 (Hsp90) without blocking ATP binding, which in turn destabilized mutant AR protein through the proteasome pathway. Intraperitoneal injection of emodin into AR-polyQ transgenic SBMA mice showed improved motor performance, alleviated body weight loss and enhanced survival rate in comparison with vehicle treated SBMA transgenic mice. Moreover, emodin treatment also attenuated spinal cord and muscle pathology, decreased the monomeric and aggregated mutant AR protein level in SBMA transgenic mice tissue. Collectively, these results indicates that emodin treatment might be a possible therapeutic approach for attenuates the manifestations of SBMA. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-235
新規抗Tau治療薬の探索のための、GFP標識Tauを用いたHigh Throughput Screeningの確立
Establishment of GFP tagged Tau based high throughput screening for the Tau modifiers discovery
*伊津野 舞佳(1)、前田 純宏(1)、岡野 栄之(1)
1. 慶應義塾大学大学院医学系研究科
*Maika Itsuno(1), Sumihiro Maeda(1), Hideyuki Okano(1)
1. Grad Sch Med, Keio Univ, Tokyo, Japan
Keyword: Neurodegenerative disorder, Tauopathy, Aging, High throughput screening
Neurodegenerative disorders characterized by Tau protein accumulation in brain are called Tauopathies. This study focuses on Alzheimer’s Disease (AD) and Frontotemporal Dementia (FTD), which affects millions of people every year. Though these are highly unmet need for drug development, most of Tau-targeted drugs have failed in clinical trials. In order to find the next candidate compounds, establishment of prompt and accurate screening system is essential. Therefore, we have established the High Throughput Screening (HTS) to find effective endogenous Tau modifiers in human neurons. We have first established induced pluripotent stem cells (iPSC) lines in which TagGFP2 is fused to the N-terminus of microtubule-associated protein tau (MAPT) gene. Moreover, we knocked in Tau A152T mutation, a MAPT gene mutation causing not only FTD but also AD. This was the only MAPT gene mutation that is genetically linked to AD. Next, we have differentiated control /A152T iPSC into neurons, using brain-enriched miRNAs and transcription factor Neurogenin 2 (Ngn2). By inserting RFP coding gene in the same plasmid with miRNAs, RFP was used as a differentiation marker. The A152T mutation increased neuronal hyper-excitation in the hiPSC-derived neurons that was reminiscent of the epileptic phenotype observed in hTau-A152T transgenic mouse model. Therefore, the reduction of the A152T Tau will be a therapeutic target to normalize abnormal Tau function. After neuronal induction, we added anti-Tau siRNA to knockdown Tau protein. Next, TagGFP2 fluorescence intensity, that is a marker for Tau protein expression, was detected using IN Cell Analyzer 6000; an automated confocal microscope. In this assay system, we have confirmed that anti-Tau siRNA effectively knockdown Tau protein. Then, we calculated Z prime (Z’), an index for the robustness of HTS, from the comparison of Tau knockdown and control neurons: From these results, we have achieved Z’ >0.5 that is high enough to carry out screenings. These indicated that the labeling endogenous Tau protein and the introduction of AD-linked MAPT gene mutation is suitable for HTS to target on endogenous pathological tau. For the next step, we will be using small compound library for HTS. Findings of the potential Tau modifier is expected. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-236
TBK1は液性因子を介して異常な単量体TDP-43を選択的に排除する。
TBK1 selectively eliminates abnormal monomeric TDP-43 via humoral factors.
*酒井 昭平(1)、大岩 康太郎(2,1)、渡邊 征爾(1)、小峯 起(1)、山中 宏二(1)
1. 名古屋大学環境医学研究所病態神経科学分野、2. 名古屋大学大学院医学系研究科神経内科
*Shohei Sakai(1), Kotaro Oiwa(2,1), Seiji Watanabe(1), Okiru Komine(1), Koji Yamanaka(1)
1. Dept Neurosci Pathobiol, RIEM, Nagoya Univ, Nagoya, Japan, 2. Dept Neurol, Grad Sch Med, Nagoya Univ, Nagoya, Japan
Keyword: amyotrophic lateral sclerosis (ALS), TDP-43, TBK1, proteostasis
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the selective death of motor neurons. TDP-43 pathology, which consists of cytosolic mislocalization, aggregation, and hyperphosphorylation of TDP-43 protein in motor neurons, is a common pathological feature in both sporadic and inherited cases of ALS. Despite identification of various ALS-causative genes, the detailed pathomechanisms linked to the TDP-43 pathology are poorly understood. TBK1 is one of notable ALS-causative genes to study the molecular basis of TDP-43 pathology because the function of TBK1 is closely related to proteostasis. However, the molecular link between TBK1 and TDP-43 mostly remains unknown due to the difficulty in reproducing TDP-43 pathology in experimental models. Recently, our group discovered that expression of the monomeric form of TDP-43 recapitulates TDP-43 pathology-like features in cells. Here, we examined the effect of TBK1 on TDP-43 protein level using its monomeric TDP-43 mutant cell model. Intriguingly, we found that a loss of TBK1 function in neuronal cells increased not wild-type (WT) but monomeric mutant TDP-43 protein level and its phosphorylated form. In contrast to suppression of TBK1, we also confirmed that overexpression of TBK1 predominantly reduced monomeric TDP-43 compared to WT. Collectively, these data suggested that TBK1 selectively eliminates abnormal monomeric TDP-43, which is prone to mislocalize and aggregate. We then explored humoral factors induced by TBK1 and discovered that conditioned medium derived from TBK1 overexpressing cells has a potential that selectively eliminates monomeric TDP-43. Fractionation of conditioned medium by ultra-filtration suggested that high-molecular-weight fractions including proteins were involved in monomeric TDP-43 clearance. Moreover, among known substrates phosphorylated by TBK1, we identified that IRF3, linked to innate immunity and cytokine secretion, is important for this phenomenon. Although elucidation of the detailed molecular mechanism of TBK1-mediated monomeric TDP-43 clearance is underway, our study suggests that TBK1 regulates proteostasis not only in cell-autonomous but also non-cell-autonomous manners, since humoral factors produced by TBK1-activated cells can affect their surrounding cells presumably through paracrine. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-237
VPS13はミトコンドリアの恒常性と神経活動において機能する
Drosophila VPS13 regulates neuronal synaptic activity and mitochondrial homeostasis
*福嶋 佳保里(1)、井下 強(1)、緒方 洵(1)、服部 信孝(1)
1. 順天堂大学大学院医学研究科
*Kahori Fukushima(1), Tsuyoshi Inoshita(1), Jun Ogata(1), Nobutaka Hattori(1)
1. Juntendo University Graduate School of Medicine
Keyword: VPS13, Parkinson's disease
Objectives:
The purpose of this study was to analyze the physiological and pathological roles of VPS13 in Drosophila neurons.
Methods:
The morphology and activity of synapses in the neuromuscular junction (NMJ) of third instar larvae were analyzed by electron microscopy and electrophysiology, respectively. The number of synaptic buttons and dopaminergic neurons were analyzed by fluorescence microscopy. Motor activity was assessed by climbing assay.
Results:
Drosophila has a single ortholog of VPS13A and VPS13C, which is named as VPS13. Electrophysiological analysis of NMJ in theVPS13-deficenet third instar larvae revealed a marked decrease in neuronal activity, while the number of synaptic buttons and cisternal structures were increased. In VPS13-deficenet aged flies, neuronal cell loss and abnormal mitochondrial morphology were observed in dopaminergic neurons of the posterior brain. Consistent with the degeneration of dopaminergic neurons, motor performance was significantly reduced in VPS13-deficenet flies. We observed the effect of human VPS13A or VPS13C reintroduction on the dopaminergic phenotypes. VPS13C rescued the altered mitochondrial morphology and dopaminergic neuron loss while VPS13A filed to do so. In contrast, motor activity was improved by both genes.
Conclusions:VPS13 plays an important role in the synaptic activity and maintenance of dopaminergic neurons in Drosophila, which may be relevant to the etiology of Parkinson's disease. Human VPS13C at least functions in regulation of mitochondrial morphology and viability of dopaminergic neurons in Drosophila. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-238
無重力飼育マウスとALSモデルマウスの脊髄神経節で発現変動する遺伝子の解析
Analysis of gene expression profiles in the dorsal root ganglion of microgravity exposed mice and ALS model mice
*吉川 雅朗(1)、松川 睦(1)、大島 秀規(2)、相澤 信(1)、志賀 隆(3)
1. 日本大学医学部機能形態学系生体構造医学分野、2. 日本大学医学部脳神経外科神経外科、3. 筑波大学医学医療系
*Masaaki Yoshikawa(1), Mutsumi Matsukawa(1), Hideki Oshima(2), Shin Aizawa(1), Takashi Shiga(3)
1. Div of Anat Sci, Dept of Funct Morphol, Nihon Univ Sch of Med, Tokyo, Japan, 2. Div of Neurosurg, Dept of Neurol Surg, Nihon Univ Sch of Med, Tokyo, Japan, 3. Fac of Med, Univ of Tsukuba, Tsukuba, Japan
Keyword: RNA-seq, DRG, Microgravity, ALS
Microgravity (MG) exposure and motor neuron disease like amyotrophic lateral sclerosis (ALS) lead to motor deficits, including muscle atrophy and neuronal activity loss. It seems to be relevant that microgravity to ALS in motor neuron degeneration. However, the role of sensory impairment in the degenerative process in MG exposure and ALS pathology is still unknown. Therefore, in the present study, to elucidate the mechanism of the sensory-motor system that is affected by each condition, we investigated the effects of MG exposure and mutant SOD1 on gene expression in various cell types of the mouse dorsal root ganglion (DRG). At first, using RNA-sequencing (RNA-seq), we examined gene expression changes in DRG of MG mice. RNA-seq analysis indicated that the expression of Fgf10 (Neurotrophic factor), Gad1, Htr2c, Trpc5 and Grm5 (nociceptive neuronal marker), Gfap (glial cell marker) was increased, and the expression of Nrtn (Neurotrophic factor), Egr3 and Cntn5 (proprioceptive neuronal marker), Csrp3 (nociceptive neuronal marker) was decreased in the DRG of MG mice. A DRG is composed of a variety of different cell types. To identify which cell types were more likely to be affected by MG or the pathogenesis of ALS, we analyzed differentially expressed genes (DEGs) with known cell-type markers derived from previous single-cell studies in the DRG of MG and SOD1 mice. DEGs were extensive in most cell types, including large neurons, medium/small neurons, satellite glial cells, Schwann cells, endothelial cells, pericytes, smooth muscle cells, macrophages, and T cells in the DRG of MG and SOD1 mice. Changes of putative large/medium neurons (NF200, TrkB, and parvalbumin groups) and cutaneous low-threshold mechanoreceptors (Mrgpb4 group) were identified as particular abnormalities in MG DRG. In the SOD1 DRG, putative medium neurons (TrkB group), cutaneous low-threshold mechanoreceptors (Mrgpb4 group), and nociceptors (TRPV1 group) were changed. These results suggest that the different DRG neurons were affected in MG and SOD1 mice. Thus, MG exposure and ALS pathology affect alterations in the DRG; possibly, various cell types contribute to motor neuron degeneration. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-239
FTLD/ALS関連C9orf72遺伝子異常伸長リピートにおけるRAN翻訳の調整メカニズム
Coordination of RAN translation in FTLD/ALS causing C9orf72-GGGGCC repeat.
*後藤 志帆(1)、森 康治(1)、山下 智子(1)、魚住 亮太(1)、宮本 哲慎(1)、近藤 志都子(1)、近江 翼(1)、河邉 有哉(1)、永井 義隆(2)、池田 学(1)
1. 大阪大学、2. 近畿大学
*Shiho GOTOH(1), Kohji Mori(1), Tomoko Yamashita(1), Ryota Uozumi(1), Tesshin Miyamoto(1), Shizuko Kondo(1), Tsubasa Omi(1), Yuya Kawabe(1), Yoshitaka Nagai(2), Manabu Ikeda(1)
1. Osaka University, 2. Kindai University
Keyword: RAN translation, C9orf72
A GGGGCC repeat expansion in intron or promoter region of C9orf72 gene is a frequent genetic cause of frontotemporal lobar degeneration (FTLD) and Amyotrophic Lateral Sclerosis (ALS). This expanded repeat is transcribed and translated into polymeric repeat proteins characterized with dipeptide motif (dipeptide repeat protein(s): DPR(s)). Such AUG codon-independent translation is called repeat associated non-AUG (RAN) translation. DPR accumulates in C9orf72-FTLD/ALS patient’ s brain and DPR toxicity has been proven in multiple disease models. Therefore, suppression of RAN translation could be one strategy for future treatment. However, detailed mechanism of RAN translation remains obscure. In this study, we explored whether and how TR1 (translation regulator 1, tentative name) modulates RAN translation in our cellular models of C9orf72-FTLD/ALS. Overexpression of TR1 enhanced RAN translation dependent expression level of most abundant poly-GA DPR. Oppositely, knockdown of TR1 decreased poly-GA expression level. Interestingly, these changes in poly-GA expression were not accompanied by corresponding changes in GGGGCC repeat RNA expression. To further investigate TR1 function, we inhibited its intrinsic activity with site directed mutagenesis. Indeed, mutated TR1 failed to enhance poly-GA expression level. Our current results suggest that TR1 positively stimulates poly-GA RAN translation though its intrinsic activity. Modulation of TR1 expression or its activity may have therapeutic potential for C9orf72-FTLD/ALS and other repeat related diseases. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-240
筋ジストロフィー犬の中枢神経症状におけるジストロフィンの役割
Role of dystrophin Dp427 in neurobehavior of canines with Duchenne Muscular Dystrophy
*竹内 絵理(1)、橋本 泰昌(1)、今村 道博(1)、菊水 健史(2)、関口 正幸(1,3)、青木 吉嗣(1)
1. 国立精神・神経医療研究センター 神経研究所 遺伝子疾患治療研究部、2. 麻布大学 獣医学部、3. 東京理科大学 薬学部薬学科
*Eri Takeuchi(1), Yasumasa Hashimoto(1), Michihiro Imamura(1), Takefumi Kikusui(2), Masayuki Sekiguchi(1,3), Yoshitsugu Aoki(1)
1. Department of Molecular Therapy, National Institute of Neuroscience, National Center of Neurology and Psychiatry, 2. School of Veterinary Medicine, Azabu University, 3. Faculty of Pharmaceutical Science, Tokyo University of Science
Keyword: dystrophin, Neurobehavioral dysfunction, Duchenne Muscular Dystrophy
Duchenne muscular dystrophy (DMD) is an X-linked neuromuscular disease that presents progressive muscle wasting. 30% of DMD shows psychiatric disorders, such as autism and high anxiety levels. DMD is caused by a mutation in the DMD gene, encoding dystrophin isoforms, including the full-length Dp427 and a shorter Dp140. Previously, we have reported that mdx mouse, a model of DMD, lacks Dp427 in muscle and brain, which exhibits freezing responses to threat, linked to the deficiency of Dp427 in the amygdala. Like mdx mice, the canine X-linked muscular dystrophy in Japan (CXMDJ) lacks Dp427 due to a point mutation in the DMD intron 6 splice acceptor site, leading to the out-of-frame skipping of exon 7. They show neurobehavioral comorbidities, however, the characterization of brain dystrophin absence and impact in brain phenotypes are still not well understood. Here, we examined the expression of Dp427 and Dp140 in the brain of CXMDJ and wild-type beagle dogs using dystrophin western blotting and immunohistochemistry. We confirmed that Dp427 was abolished from the CXMDJ brain. We also performed a series of behavior tests and found that CXMDJ showed anxiety-like behavior probably due to a lack of Dp427. Our results suggest that CXMDJ shows similar increased fear responses to threat to the mdx mouse. The brain symptoms might be valuable for brain-targeted dystrophin-restoration therapies. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-241
神経炎症がパーキンソン病モデルマウスのα-シヌクレイン病理を増強する。
Neuroinflammation potentiates α-synuclein pathology in a mouse model of Parkinson's disease.
*早川 英規(1)、馬場 孝輔(1,2)、佐木山 裕史(1)、Choong Chi-Jing(1)、木村 康義(1)、池中 建介(1)、Aguirre Cesar (1)、奥野 龍禎(1)、長野 清一(1,3)、永井 義隆(4)、望月 秀樹(1)
1. 大阪大学大学院 医学系研究科 神経内科学、2. 富山大学 学術研究部医学系 脳神経内科、3. 大阪大学大学院 医学系研究科 神経難病認知症探索治療学、4. 近畿大学大学院 医学研究科 脳神経内科
*Hideki Hayakawa(1), Kousuke Baba(1,2), Hiroshi Sakiyama(1), Chi-Jing Choong(1), Yasuyoshi Kimura(1), Kensuke Ikenaka (1), Cesar Aguirre(1), Tatsusada Okuno(1), Seiichi Nagano(1,3), Yoshitaka Nagai(4), Hideki Mochizuki(1)
1. Department of Neurology, Osaka University Graduate School of Medicine, 2. Department of Neurology, Faculty of Medicine, University of Toyama, 3. Department of Neurotherapeutics, Osaka University Graduate School of Medicine, 4. Department of Neurology, Faculty of Medicine, Kindai University
Keyword: Parkinson's disease, α-synuclein, Neuroinflammation
Objective: α-synuclein (αsyn) is a major component of Lewy bodies, a pathological hallmark of Parkinson's disease (PD), and its genetic mutations cause familial PD. In neurodegenerative diseases such as PD and Alzheimer's disease, the presence of Tumor necrosis factor-α and Interleukin (IL)-1-positive microglia involved in neuroinflammation has long been recognized as important, but the exact role of the immune response in disease progression is not clear. Although the relationship between αsyn and microglia has been discussed from the perspective of neurodegeneration and neuronal damage, the propagation and aggregation of αsyn have not been clearly demonstrated. In this study, we analyzed the molecular mechanisms of neuroinflammation and αsyn propagation and aggregation induced by activated microglia using αsyn fibril-injected mice. Methods: αsyn fibrils were injected into the substantia nigra of both wild-type and ASC (apoptosis-associated speck-like protein containing caspase recruitment domain) KO mice. ASC forms a complex called the NLRP3 inflammasome, which induces the maturation and production of the pro-inflammatory cytokines IL-1β and IL-18 via the protease Caspase-1 (Misawa T, et al. Nat Immunol. 2013). Therefore, in ASC KO mice, microglial activation and induction of inflammatory cytokine production are suppressed. We analyzed the pathology of nigral dopaminergic neuronal loss, αsyn aggregate formation, and microglial status and distribution in the brains of αsyn fibril-injected wild-type and ASC KO mice.
Result: ASC KO mice showed reduced phosphorylated-αsyn aggregate formation and dopamine cell loss in the substantia nigra compared to wild-type mice, but no significant difference in the propagation of αsyn to the cortex or striatum. Conclusion: These results suggest that neuroinflammation is involved in the formation of αsyn aggregates and the loss of dopaminergic neurons, and hence plays an important role in the progression of synucleinopathy. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-242
FTLDモデルマーモセットを用いた認知機能評価および摂食行動評価の確立
Establishment of cognitive function assessment and eating behavior assessment using FTLD model marmosets
*金 侑璃(1)、岩田（遠藤） 邦幸(1,2)、畑 純一(3)、力武 聖月(3)、勝野 雅央(1)、祖父江 元(4)、中村 克樹(5)、石垣 診祐(1,6)
1. 名古屋大学大学院医学系研究科神経内科、2. 名古屋大学大学院医学系研究科精神科、3. 東京都立大学、4. 愛知医科大学、5. 京都大学 霊長類研究所、6. 名古屋大学 脳とこころの研究センター
*Yuri Kim(1), Kuniyuki Iwata(Endo)(1,2), Junichi Hata(3), Mitsuki Rikitake(3), Masahisa Katsuno(1), Gen Sobue(4), Katsuki Nakamura(5), Shinsuke Ishigaki(1,6)
1. Department of Neurology, Nagoya University Graduate School of Medicine., 2. Department of Psychiatry, Nagoya University Graduate School of Medicine., 3. Tokyo Metropolitan University, 4. Aichi Medical University, 5. Primate Research Institute, Kyoto University, 6. Brain and Mind Research Center, Nagoya University
Keyword: Frontotemporal Lobar Degeneration (FTLD), marmoset, Fused in Sarcoma, ALS
Objective: This study aimed to establish early biomarkers for neurological diseases such as frontotemporal lobar degeneration (FTLD) by creating model marmosets and conducting cognitive and behavioral experiments. Background: FTLD is a type of dementia with distinct behavioral abnormalities including decreased exploratory behavior and abnormal eating behavior. Altered decision-making has been observed in FTLD/ALS patients in probabilistic reversal learning (PRL) tasks. Fused in sarcoma (FUS), an RNA-binding protein, is involved in the pathogenesis of ALS as well as FTLD. In this study, we established a marmoset model of FUS silencing by stereotactic injection (FUS-KD marmoset) and investigated their behavioral alterations. Methods: We designed shRNAs against the marmoset FUS gene and generated an AAV9 virus encoding the most effective shRNA against FUS (shFUS). The AAV encoding shFUS (AAV-shFUS) was introduced into the bilateral caudate heads of young adult marmosets, whereas AAV encoding a control shRNA was injected as controls. Next, we developed a tablet-based PRL task system for marmosets to investigate decision-making abnormalities. In order to evaluate the feeding behavior of model marmosets, we first developed a method to quantify the feeding behaviors of marmosets. MRI scans of the brains of marmosets were performed to examine the changes in neural connectivity in the brain caused by FUS-KD. Results: We found that the FUS-KD marmoset mimicked FTLD-like features characterized by reduced selectivity in the PRL task 6 months after FUS silencing. MRI analysis revealed that that the neural connectivity between the PFC and caudate nucleus was reduced by FUS silencing in marmosets. Conclusion: The FUS-KD marmoset model can be considered as a suitable animal model for the development of translatable indices with humans. We will continue to analyze macroscopic network changes by MRI, functional network changes by fMRI, and probability reversal learning tasks, as well as feeding behavior experiments. In order to establish them as a disease biomarker battery, we would clarify the chronological changes in these indices in the near future. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-243
ラット小脳由来正常アストロサイトにおけるメチル水銀誘発毒性における酸化ストレスの関与
Involvement of oxidative stress in methylmercury-induced toxicity in cultured normal rat cerebellar astrocytes
*佐々木 翔斗(1)、根岸 隆之(1,2)、都築 孝允(2)、湯川 和典(1,2)
1. 名城大学大学院薬学研究科、2. 名城大学薬学部
*Shoto Sasaki(1), Takayuki Negishi (1,2), Takamasa Tsuzuki(2), Kazunori Yukawa(1,2)
1. Graduate school of Pharmacy, Meijo University, 2. Faculty of Pharmacy, Meijo University
Keyword: Methylmercury, Astrocytes, Oxidative stress
Methylmercury (MeHg) is known as the causative agent of Minamata disease, which accumulates in brain and damages nerve cells. We previously reported that exposure to 2.5 µM MeHg for 96 h caused activation of MAPK and transcription factors in cultured normal rat cerebellar astrocytes (NRA). In the present study, we investigated the effects of antioxidants (Trolox, N-acetyl-cysteine (NAC), and glutathione (GSH)) on MeHg toxicity in NRA to understand its biological mechanism and propose possible suppression methods. NRA from Wistar rat cerebellum were seeded after the last passage and cultured in FBS-containing medium for 48 h and in the serum-free medium for 48 h. Cell viability was evaluated in NRA exposed to 0, 2, or 4 µM MeHg and 0–200 µM Trolox, 0–4 mM NAC, or 0–4 mM GSH for 96 h. Protein expression was evaluated by Western blot in NRA exposed to 2 µM MeHg and 100 and 200 µM Trolox, 2 and 4 mM NAC for 96 h. In addition, intracellular ROS was measured in NRA exposed to 0–10 µM MeHg for 96 h. Cell viability was increased by exposure to 2 µM MeHg for 96 h, which was suppressed by simultaneous exposure to 100 or 200 µM Trolox or 2 or 4 mM NAC. Unexpectedly, simultaneous exposure to 2 µM MeHg and 0.5–4 mM GSH in NRA led to remarkable cell death. Exposure to 4 µM MeHg for 96 h caused cell death, and exposure to 100 and 200 µM Trolox, 0.5–4 mM NAC, or 4 mM GSH suppressed the MeHg-induced cell death. Exposure to 2 µM MeHg for 96 h increased phosphorylation of MAPK (ERK1/2, p38MAPK, SAPK/JNK), expression and phosphorylation of transcription factors (c-Jun, c-Fos), and expression of Hsp70, while it decreased expression of Nrf2, which was improved by simultaneous exposure to 100 or 200 µM Trolox or 2 mM NAC. Exposure to 2 µM MeHg for 96 h increased expression of HO-1, which was suppressed by exposure to 2 mM NAC and further increased by exposure to 100 or 200 µM Trolox. In addition, exposure to 2 µM MeHg for 96 h increased intracellular ROS. While the ROS production was increased by MeHg exposure, an abnormality occurred in the expression of Hsp70, Nrf2, and HO-1 and they were improved by Trolox or NAC mostly, which implies that MeHg in astrocytes might induce the disruption of cellular ROS balance in NRA. On the other hand, GSH exacerbated the adverse effects of 2 µM MeHg, suggesting that well-known antioxidants could not necessarily suppress the effects of MeHg and that toxicity mechanism of MeHg might differ qualitatively depending on the MeHg concentration. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-244
Neuroprotection against Parkinson’s disease through the attenuation of mitochondrial dysfunction and neuroinflammation mediated apoptosis by Chlorogenic acid
*Saumitra Singh(1), Surya Singh(1)
1. Banaras Hindu University
Keyword: Parkinson's disease, Chlorogenic acid, Mitochondrial dysfunction
Parkinson’s disease (PD) pathology is based on neuroinflammation, oxidative stress and mitochondrial dynamics, which act as major pillars in disease pathogenesis. Although the therapies work good for regressing the clinical symptoms of the disease, but unable to control the loss of the dopaminergic neurons. In this study, the effectiveness of chlorogenic acid (CGA) against neuropathological implications of Parkinson’s disease in 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) induced Parkinsonian mice has been explored. Henceforth, significant elevation in tyrosine hydroxylase (TH) expression and reoccurrence of motor coordination and antioxidant defence was observed within the nigrostriatal region of the Parkinsonian mouse model when compared to MPTP-injected mice group. Moreover, nuclear factor-kB (NF-κB) expression was also reduced in CGA treated Parkinsonian mice; indicating it’s effectiveness in reducing the neuroinflammation. Also, the expression of proinflammatory mediators such as tumor necrosis factor-α and interleukin (IL)-1β was inhibited and expression of anti-inflammatory cytokine IL-10 was also enhanced on CGA supplication in Parkinsonian mice. Furthermore, astrocyte activation attributed by GFAP expression was also found to be reduced, suggesting the neuroprotective role of CGA in PD. The compound restored the fall in activity of mitochondrial complexes I, IV, V and membrane potential by ameliorating the level of superoxide dismutase and mitochondrial glutathione in the midbrain of PD mouse. The proapoptotic markers including Bax and caspase-3 were inhibited and the anti-apoptotic protein, Bcl-2 was elevated on CGA administration, thereby preventing the MPTP-mediated apoptotic cascade. CGA has also increased the phosphorylation of Akt and GSK3β, suggesting it’s neuroprotective effects. Thus, the present study put forward CGA as a novel therapeutic avenue for possible clinical intervention in PD therapy. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-245
Longitudinal analysis of primate progressive synucleinopathies model revealed slowing in saccade reaction time.
*Wajd Amly(1), Chih-Yang Chen(1,2), Tadashi Isa(1,2,3)
1. Department of Neuroscience, Graduate school of Medicine, Kyoto University, 2. Institute for the Advanced Study of Human Biology (WPI-ASHBI), Kyoto University, 3. Human Brain Research Center, Graduate School of Medicine, Kyoto University
Keyword: saccades, α-Synuclein, oculomotor
The oculomotor system not just intergrades the visual input and the motor output, but also reflects the cognitive state and thus, is widely used to probe brain functions and neural disorders in humans. Recent studies suggested that the α-Synuclein fibrils can lead to synucleinopathies, including Parkinson’s disease (PD) and dementia with Lewy bodies. However, whether the animal models show similar oculomotor deficits to human patients were not tested. Being motivated by this, we trained three common marmosets on oculomotor tasks and observed their saccade behaviour before and after α-Synuclein fibril injection. To approach this, we trained our marmosets on tasks related to both internally and externally driven saccades, the oculomotor delayed response task (ODR) and step and gap tasks, respectively, because the neurons respond to these tasks differently in brain structures related to oculomotor control. After collecting 6 months baseline data, we injected α-Synuclein fibrils into the olfactory bulb (OB) bilaterally, following Braak’s dual hit hypothesis for PD. With a longitudinal follow-up, we found that saccade reaction time (SRT) in gap task gradually got slower with the time after α-Synuclein injection. The SRT in ODR first got slower 3 months after injection but got faster later on. We also found that SRT in step saccade task was less affected than in gap task. This suggested cortical generation of saccade may be affected by α-Synuclein. Furthermore, we found that the level of attention and arousal of the marmoset decreased with the progression of the disease in the ODR task. We further did through rough analysis on microsaccades and didn’t find any change in microsaccade frequency or direction, suggesting bottom-up saliency was not affected. In conclusion, the OB α-Synuclein disease model showed a change in SRT and attention level as synucleinopathies progressed with time. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-246
胎児期PGD2受容体アゴニスト暴露は社会性行動や神経細胞形態に影響を及ぼす
Prenatal exposure of the PGD2 receptor agonist effects on social communication and neuronal morphology in male offspring.
*早田 敦子(1,2)、菊池 泰河(2)、豊田 博紀(3)、毛利 育子(4)、橘 雅弥(1,4)、加藤 隆史(1,3)、谷池 雅子(4)、橋本 均(1,2,5,6,7)
1. 大阪大学大学院連合小児発達学研究科 附属子どものこころの分子統御機構研究センター、2. 大阪大学大学院薬学研究科 、3. 大阪大学大学院歯学研究科、4. 大阪大学大学院連合小児発達学研究科、5. 大阪大学大学院医学研究科、6. 大阪大学データビリティフロンティア機構、7. 大阪大学先導的学際研究機構
*Atsuko Hayata-Takano(1,2), Taiga Kikuchi(2), Hiroki Toyoda(3), Ikuko Mohri(4), Masaya Tachibana(1,4), Takafumi Kato(1,3), Masako Taniike(4), Hitoshi Hashimoto(1,2,5,6,7)
1. Mol Res Center for Child Mental Develop, United Grad Sch Child Development, Osaka Univ, Osaka, Japan, 2. Grad Sch Pharmaceutical Sci, Osaka Univ, Osaka, Japan, 3. Grad Sch Dent, Osaka Univ, Osaka, Japan, 4. United Grad Sch Child Development, Osaka Univ, Osaka, Japan, 5. Grad Sch Med, Osaka Univ, Osaka, Japan, 6. Inst Datability Sci, Osaka Univ, Osaka, Japan, 7. Inst Open Transdiscip Res Initiatives, Osaka Univ, Osaka, Japan
Keyword: neurodevelopmental disorders, Prostaglandin, neuronal morphology
Accumulating evidence from human and animal model studies suggests that neurodevelopmental disorders such as autism spectrum disorders and schizophrenia can be linked to early life inflammation. Prostaglandin D2 (PGD2) is an inflammatory mediator produced by the activities of cyclooxygenase (COX) and either hematopoietic PGD synthase (H-PGDS) or lipocalin-type PGD synthase (L-PGDS). In the central nervous system (CNS), PGD2 plays an important role as a neuromodulator in the regulation of the sleep-wake cycle, body temperature, hormone release, and pain responses via the G protein–coupled receptors DP1 and DP2/CRTH2. Recently, we found that the expression level of H-PGDS in the frontal lobe was higher in autistic human brains than control human brains, and the DP1 agonist increased the number of primary dendrites and dendritic spines in the mouse primary cultured cortical neurons at 21 days in vitro. These findings indicate that PGD2/DP1 signaling might be involved in neuronal development. However, the role of PGD2-DP1 signaling in neuronal development remains largely unknown. In this study, we investigated that the behavioral and cellular effects of DP1 agonist injection on day 12.5 of mouse pregnancy. A single maternal injection of BW 245C, a specific DP1 agonist, reduced the number of ultrasonic calls at postnatal day 7 and social behavior in the juvenile and adult male offspring. In addition, prenatal exposure to BW 245C caused alterations in the dendritic morphology and the number of dendritic synapses in the prefrontal cortex. These results suggest that DP1 signaling is implicated in neuronal development and the over-activation of PGD2-DP1 signaling during early development could be a risk factor for neurodevelopmental disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-247
オキシトシン受容体発現細胞を可視化・操作するための新たなゲノム編集マウス
Targeting neurons with functional oxytocin receptors: A novel set of simple knock-in mouse lines for oxytocin receptor visualization and manipulation
*井上（上野） 由紀子(1)、三輪 秀樹(2)、堀 啓(3)、金子 涼輔(4)、森本 由起(1)、小池 絵里子(1)、浅見 淳子(1)、上條 諭志(2)、山田 光彦(2)、星野 幹雄(3)、井上 高良(1)
1. 国立精神・神経医療研究センター疾病研究第６部、2. 国立精神・神経医療研究センター精神薬理研究部、3. 国立精神・神経医療研究センター病態生化学研究部、4. 大阪大学大学院生命機能研究科時空生物学
*Yukiko U. Inoue(1), Hideki Miwa(2), Kei Hori(3), Ryosuke Kaneko(4), Yuki Morimoto(1), Eriko Koike(1), Junko Asami(1), Satoshi Kamijo(2), Mitsuhiko Yamada(2), Mikio Hoshino(3), Takayoshi Inoue(1)
1. Dep of Biochem and Cellular Biol, National Institute of Neurosci, Tokyo, Japan, 2. Dep of Neuropsychopharmacology, National Institute of Mental Health, Tokyo, Japan, 3. Dep of Biochem and Cellular Biol, National Institute of Neurosci, Tokyo, Japan, 4. KOKORO-Biol, Lab for Integrated Biol, Grad Sch of Frontier Biosci, Osaka Univ, Osaka, Japan
Keyword: Oxytocin, Oxytocin receptor, genome editing, knock-in mice
The neuropeptide oxytocin (Oxt) plays important roles in modulating social behaviors. Oxytocin receptor (Oxtr) is abundantly expressed in the brain and its relationship to socio-behavioral controls has been extensively studied using mouse brains. Several genetic tools to visualize and/or manipulate Oxtr-expressing cells, such as fluorescent reporters and Cre recombinase drivers, have been generated by ES-cell based gene targeting or bacterial artificial chromosome (BAC) transgenesis. However, these mouse lines displayed some differences in their Oxtr expression profiles probably due to the complex context and integrity of their genomic configurations in each line. Here we apply our sophisticated genome-editing techniques to the Oxtr locus, systematically generating a series of knock-in mouse lines, in which its endogenous transcriptional regulations are intactly preserved and evaluate their expression profiles to ensure the reliability of our new tools. We employ the epitope tagging strategy, with which C-terminally fused tags can be detected by highly specific antibodies, to successfully visualize the Oxtr protein distribution on the neural membrane with super-resolution imaging for the first time. By utilizing T2A self-cleaving peptide sequences, we also induce proper expressions of tdTomato reporter, codon-improved Cre recombinase, and spatiotemporally inducible Cre-ERT2 in Oxtr-expressing neurons. Electrophysiological recordings from tdTomato-positive cells in the reporter mice support the validity of our tool design. Retro-orbital injections of AAV-PHP.eB vector into the Cre line further enable visualization of recombinase activities in the appropriate brain regions. Moreover, the first-time Cre-ERT2 line drives Cre-mediated recombination in a spatiotemporally controlled manner upon tamoxifen administration. These tools thus provide excellent resource for future functional studies in Oxt-responsive neurons and should prove of broad interest in the field. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-248
Rho活性化因子PLEKHG2の機能欠損は神経発達を障害する
Impaired function of PLEKHG2, a Rho-guanine nucleotide-exchange factor, disrupts corticogenesis in neurodevelopmental phenotypes
*西川 将司(1)、伊東 秀記(1)、田畑 秀典(1)、永田 浩一(1)
1. 愛知県医療療育総合センター発達障害研究所
*Masashi Nishikawa(1), Hidenori Ito(1), Hidenori Tabata(1), Koh-ichi Ngata(1)
1. Institute for Developmental Research, Aichi Developmental Disability Center
Keyword: Rho family small GTPase, RhoGEF, Brain development, Neurodevelopmental disorders
While homozygosity of p.Arg204Trp variation in PLEKHG2, encoding a Rho family-specific guanine nucleotide-exchange factor, is responsible for microcephaly with intellectual disability, the role of PLEKHG2 in the neurodevelopmental process is unknown. In this study, we analyzed the role of mouse Plekhg2 in the cortical development both in vitro and in vivo. The p.Arg200Trp variant in mouse Plekhg2 (Plekhg2-RW) corresponding to the human p.Arg204Trp showed decreased guanine nucleotide-exchange activity for Rac1, Rac3, and Cdc42. Acute knockdown of Plekhg2 with an in utero electroporation-mediated gene transfer method did not affect migration of excitatory neurons during corticogenesis. On the other hand, silencing of Plekhg2 delayed dendritic arbor formation at postnatal day 7 (P7), perhaps because of impaired Rac/Cdc42 and p21-activated kinase 1 signaling pathway. The phenotype was rescued by an RNAi-resistant version of wildtype Plekhg2, but not by Plekhg2-RW. Axon pathfinding was also impaired in vitro and in vivo in Plekhg2-deficient cortical neurons. At P14, knockdown of Plekhg2 was observed to cause defects in dendritic spine morphology. From the results that Plekhg2-deficiency caused morphological defects in axons, dendrites, and synapses during brain development, PLEKHG2 was found to be crucial for brain development. Also, its functional defects are responsible for neurodevelopmental disorders with microcephaly and ID in the patients with the p.Arg204Trp variation in PLEKHG2. As for an underlying mechanism in the clinical phenotypes, disrupted actin regulation caused by aberrant Rac/Cdc42-PAK1 signaling is most likely to be a core event; hampered actin reorganization gives rise to defective neuron morphology including abrogated neurite growth and spine formation, which are relevant to pathophysiology of ID and microcephaly in “PLEKHG2 disease”. Further intensive analysis of the molecular machinery should contribute to a better understanding of the pathophysiology of the disease. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-249
低分子量G蛋白質RAC3変異による神経発達障害の分子病態メカニズムの解明
Variant-specific changes in RAC3 function disrupt corticogenesis in neurodevelopmental phenotypes
*永田 浩一(1)、西川 将司(1)、伊東 秀記(1)
1. 愛知県医療療育総合センター 発達障害研究所
*Koh-ichi Nagata(1), Masashi Nishikawa(1), Hidenori Ito(1)
1. Institute for Developmental Research, Aichi Developmental Disability Center
Keyword: RAC3, small GTPase, microcephaly, intellectual disability
Variants in RAC3, encoding a small GTPase RAC3 which is critical for the regulation of actin cytoskeleton and intracellular signal transduction, are associated with a rare neurodevelopmental disorder with structural brain anomalies and facial dysmorphism. We investigated a cohort of 10 unrelated participants presenting with global psychomotor delay, hypotonia, behavioral disturbances, stereotyped movements, dysmorphic features, seizures, and musculoskeletal abnormalities. MRI of brain revealed a complex pattern of variable brain malformations, including callosal abnormalities, white matter thinning, gray matter heterotopia, polymicrogyria/dysgyria, brainstem anomalies, and cerebellar dysplasia. These patients harbored eight distinct de novo RAC3 variants, including six novel variants (NM_005052.3): c.34G>C p.G12R, c.179G>A p.G60D, c.186_188delGGA p.E62del, c.187G>A p.D63N, c.191A>G p.Y64C, and c.348G>C p.K116N. We then examined the pathophysiological significance of these novel and previously reported pathogenic variants p.P29L, p.P34R, p.A59G, p.Q61L, and p.E62K. In vitro analyses revealed that all tested RAC3 variants were biochemically and biologically active to variable extent, and exhibited a spectrum of different affinities to downstream effectors including p21-activated kinase 1(PAK1). We then focused on the four variants p.Q61L, p.E62del, p.D63N, and p.Y64C, in the Switch II region, which is essential for the biochemical activity of small GTPases and also a variation hot spot common to other Rho family genes, RAC1 and CDC42. Acute expression of the four variants in embryonic mouse brain using in utero electroporation caused defects in cortical neuron morphology and migration ending up with cluster formation during corticogenesis. Notably, defective migration by p.E62del, p.D63N, and p.Y64C were rescued by a dominant negative version of PAK1. Our results indicate that RAC3 variants result in morphological and functional defects in cortical neurons during brain development through variant-specific mechanisms, eventually leading to heterogeneous neurodevelopmental phenotypes. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-250
オートファジー機能低下を伴うASDモデル動物の電気生理学的解析
Electrophysiological analysis of ASD model mice with impaired autophagy
*大野 萌馨(1,2)、若月 修二(1)、荒木 敏之(1,2)
1. 国立精神・神経医療研究センター神経研究所、2. 東京農工大学工学府
*Moeka Ohno(1,2), Shuji Wakatsuki(1), Toshiyuki Araki(1,2)
1. Natl Inst Neurosci, NCNP, Tokyo, Japan, 2. Grad Sch Eng, TUAT, Tokyo, Japan
Keyword: autophagy, ASD
Autism Spectrum Disorder (ASD) is a neurodevelopmental disorder characterized by impaired social communication and repetitive behaviors. In recent years, it has become clear that selective proteolysis regulates neural functions, and that disruption of proteolytic mechanisms is involved in the pathogenesis of neurodevelopmental disorders, including ASD. However, it remains unclear how the disruption of proteolytic mechanisms affects the formation and development of neural networks. Previously, we have shown that Na+-H+exchanger 5 gene-deficient (NHE5 KO) mice exhibit ASD-like behavioral abnormalities, which can be rescued by enhancing autophagy function. In the present study, we performed electrophysiological analysis to investigate the neurological dysfunction under NHE5 KO and to clarify its relationship with autophagy dysfunction. Using the whole-cell patch clamp technique, we measured and analyzed the amplitude of mEPSC and mIPSC, the Paired-Pulse ratio (PPR), and the reversal potential of AMPA receptors in CA1 pyramidal cells of wild-type (C57BL/6J) and NHE5 KO mice (male, 4-6 weeks old). We found that the amplitude of mEPSC was reduced while that of mIPSC was increased in pyramidal cells of NHE5 KO mice. This resulted in a decrease in the mEPSC/mIPSC ratio of amplitude. On the other hand, the PPR and the reversal potential of AMPA receptors were not changed compared to that of the wild type mice, respectively. These findings suggested that the CA1 pyramidal cells of NHE5 KO mice exhibit the disruption of E-I balance. Furthermore, the decrease of mEPSC amplitude might not come from the change of release amount of transmitter or AMPA receptor reversal potential caused by the change of ion concentration. Considering our previous result that the length of PSD beneath postsynaptic membrane measured on electron microscopic images of synapses of NHE5 KO mice was shorter than that of the wild type mice, our current findings suggest that morphological changes of synapses contribute to the reduction of mEPSC amplitude. Further studies are needed to examine whether enhancement of autophagy function might rescue electrophysiological phenotype of NHE5 KO mice. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-251
結節性硬化症モデルマウスにおける、アストロサイトの活性化に起因した記憶障害メカニズム
Reactive astrocyte-mediated mechanisms of memory disorder in tubular sclerosis complex model mouse
*島田 忠之(1)、杉浦 弘子(1)、山形 要人(1,2)
1. 東京都医学総合研究所こどもの脳プロジェクト、2. 高田西城病院
*Tadayuki Shimada(1), Hiroko Sugiura(1), Kanato Yamagata(1,2)
1. Child Brain Project, Tokyo Metropolitan Institute of Medical Science, 2. Takada Nishishiro Hospital
Keyword: Social memory, Tubular sclerosis complex, Reactive astrocyte, Glia-neuron crosstalk
Tuberous sclerosis complex (TSC) patients harbor hamartomas in the brain and other organs. The neuropsychiatric symptoms of TSC patients include refractory epilepsy, autism spectral disorders and mental retardation. One of the responsible genes for TSC is Tsc1. To investigate if the brain astrocytes contribute to the neuropsychiatric symptoms of TSC patients, we developed astrocyte-specific Tsc1 knockout mice (cKO mouse) as a TSC model mouse and examined their phenotypes. Immunohistochemical analyses showed that GFAP-highly-positive cells, reactive astorsytes, were increased in the brain, especially in the hippocampus in cKO mice. In addition, the malformed dendritic spines ware observed in dentate gyrus in hippocampi, where one of the most severe gliosis was observed. Moreover, three-chamber test analysis revealed that the cKO mice showed impaired social memory, suggesting that the mutant mice show autistic behavior. Deletion of Tsc1 activates small G-protein, Rheb, and expression of syntenin protein is upregulated in the active-Rheb-dependent-manner. Therefore, we examined whether this signaling pathway is involved in the induction of abnormal social memory in the cKO mouse. Treatment with Rheb inhibitors restored gliosis, spine malformation, syntenin over-expression, and impaired social memory in cKO mouse. In addition, Tsc1/syntenin double cKO mice showed recovery of gliosis and improved social memory. These results suggested that activation of the Tsc/Rheb/syntenin signaling in the astrocytes could induce astrogliosis, affecting the spine morphology by the crosstalk between the astrocytes and neurons. Resulting neural dysfunction would develop abnormal social behaviors observed in a mouse model of TSC. Finally, we found that phosphorylation of NF-κB is regulated by the Rheb/syntenin in cKO mice astrocyte. NF-κB-dependent gene expression in the reactive astrocyte could affect neuronal morphology and function in TSC mouse. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-252
神経発達症様の行動を示す齧歯類において軸索起始部の長さが変化する
Length impairments of the axon initial segment in rodent models of attention-deficit hyperactivity disorder and autism spectrum disorder
*田 小叶(2)、臼井 紀好(1,2,3)、張替 若菜(2)、三好 耕(2)、篠田 晃(4)、田中 潤也(5)、島田 昌一(1,2,3)、片山 泰一(2)、吉村 武(2)
1. 大阪大学大学院医学系研究科、2. 大阪大学大学院連合小児発達学研究科、3. 大阪精神医療センター・こころの科学、4. 山口大学大学院医学系研究科、5. 愛媛大学大学院医学系研究科
*Xiaoye Tian(2), Noriyoshi Usui(1,2,3), Wakana Harigai(2), Ko Miyoshi(2), Koh Shinoda(4), Junya Tanaka(5), Shoichi Shimada(1,2,3), Taiichi Katayama(2), Takeshi Yoshimura(2)
1. Grad Sch Med, Osaka Univ, Suita, Japan, 2. United Grad Sch Child Development, Osaka Univ, Suita, Japan, 3. Addiction Research Unit, Osaka Psychiatric Research Center, Osaka Psychiatric Medical Center, Hirakata, Japan, 4. Grad Sch Med, Yamaguchi Univ, Ube, Japan , 5. Grad Sch Med, Ehime Univ, Toon, Japan
Keyword: Axon initial segment, Neurodevelopmental disorder, Attention-deficit hyperactivity disorder, Autism spectrum disorder
The axon initial segment (AIS) is a structural neuronal compartment of the proximal axon that plays key roles in sodium channel clustering, action potential initiation, and signal propagation of neuronal outputs. Recent studies demonstrate that the AIS geometry (i.e., length or position along the axon) can be dynamic and plastic in response to normal developmental or pathological activities. The computational model study suggests that reducing the AIS length by a few micrometers could result in increased interspike interval and decreased neuronal excitability. Hence, abnormalities in the length of AIS are expected to alter neuronal excitability. Mutations in constitutive genes of the AIS, such as ANK3, have been identified in patients with neurodevelopmental disorders. Nevertheless, morphological changes in the AIS in neurodevelopmental disorders have not been characterized. In this study, we investigated the length of the AIS in rodent models of attention-deficit hyperactivity disorder and autism spectrum disorder. We observed abnormalities in AIS length in both animal models. In mouse and rat models of attention-deficit hyperactivity disorder, we observed shorter AIS length in specific brain regions. In addition, we found shorter lengths of Pan-Nav staining in attention-deficit hyperactivity disorder model rats. Further, we observed shortening of AIS lengths in autism spectrum disorder model mice. These results suggest that impairments in AIS length are common phenomena in neurodevelopmental disorders such as attention-deficit hyperactivity disorder and autism spectrum disorder and may be conserved across species. Our findings provide novel insight into the potential contribution of the AIS to the pathophysiology and pathogenesis of neurodevelopmental disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-253
Gad1ヘテロ接合体への胎生期ストレスは神経発達及び行動関連遺伝子にエピジェネティックの制御異常をもたらす
Gad1 heterozygotes subjected to prenatal stress undergo epigenetic dysregulation of genes perturbing neurodevelopment and behavioral phenotypes.
*王 天英(1,2)、Adya Saran Sinha(1)、武藤 弘樹(1)、柳川 右千夫(3)、河合 智子(4)、秦 健一郎(4)、福田 敦夫(1)
1. 浜松医大・神経生理学講座、2. 浜松医大・生体機能イメージング研究室、3. 群馬大院・脳神経発達統御・遺伝発達行動、4. 国立成育医療研究センター研究所・周産期病態研究部
*Tianying Wang(1,2), Adya Saran Sinha(1), Hiroki Mutoh(1), Yuchio Yanagawa(3), Tomoko Kawai(4), Kenichiro Hata(4), Atsuo Fukuda(1)
1. Dept Neurophysiol., Hamamatsu Univ. Sch. Med., Hamamatsu, Japan, 2. Dept Biofunc imaging.,Hamamatsu Univ. Sch. Med., Hamamatsu, Japan, 3. Dept. Genet Behav Neurosci., Gunma Univ. Grad. Sch. Med., Maebashi, Japan, 4. Dept. Maternal-Fetal Biol., Natl Res Inst Child Health Dev., Tokyo, Japan
Keyword: Gad1 mutation, prenatal stress, epigenetic, neurodevelopment
Exposure to prenatal stress (PS) and mutations in Gad1, which encodes the GABA synthesizing enzyme glutamate decarboxylase (GAD) 67, are both risk factors for psychiatric disorders. Using GAD67-GFP knock-in heterozygous (HT) mice subjected to PS from embryonic day 15.0 to 17.5, we previously reported disruption of GABAergic neurogenesis in the MGE. Postnatally, the density of parvalbumin (PV)-positive GABAergic interneurons was significantly decreased in the mPFC of HT-PS mice, which is a common trait shared by different psychiatric diseases. When assessing the function of interneurons in the mPFC of HT-PS, we found that the excitability of the inhibitory network was altered in HT-PS mice. These results suggested that these two key genetic and environmental susceptibility factors could specifically disturb GABAergic system. By contrast, these findings were not observed in wild type offspring. In this study using a genetic risk factor (Gad1 heterozygosity) with prenatal stress, an environmental risk factor, we evaluated effect on epigenetic programming and subsequent behavioral deficits. Differentially expressed genes (DEGs) detected by high-throughput microarray expression were functionally associated with neurogenesis and behavior in HT-PS mice. Some DEGs were strongly correlated to their altered epigenetic status as validated by real-time PCR. Among these genes, Lhx8, Isl1 and Ngfr are associated with transcriptional regulation of cell fate in MGE precursor cells and promoting cholinergic genes expression. Consistent with increase in the expression of cholinergic related genes, we found the cholinergic terminals in the mPFC of HT-PS mice were significantly increased. Further study showed increase in spontaneous GABAergic network activity in mPFC of HT-PS mice was blocked by muscarinic receptor antagonist atropine, suggesting the possibility of increments of cholinergic excitatory neuromodulation onto the GABAergic neurons. In addition, we examined the behavioral phenotypes and found that HT-PS mice demonstrated significant deficits in sensorimotor gating, cognitive processes, social engagement, without change in anxiety and exploratory behaviors. Finally, electrocorticogram recording showed reduction in power spectrum density at gamma-frequency range in mPFC of HT-PS mice, indicating GABAergic dysfunction could underlie the behavioral deficit. These findings may provide new insights into mechanisms of the pathogenesis of psychiatric disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-254
マイクロRNAによるクラッベ病モデルマウスオリゴデンドロサイトの病態改善効果
miR-219 restored oligodendrocyte development impaired in a mouse model of Krabbe disease.
*稲村 直子(1)、郷 慎司(2)、渡辺 昂(2)、高瀬 弘嗣(3)、中山 敦雄(1,4)、竹林 浩秀(5)、松田 純子(2)、榎戸 靖(1)
1. 愛知県医療療育総合センター 発達障害研究所 、2. 川崎医大・病態代謝学、3. 名市大・医学研究科、4. 名大・医学系研究科、5. 新潟大・医歯学総合研究科
*Naoko Inamura(1), Shinji Go(2), Takashi Watanabe(2), Hiroshi Takase(3), Atsuo Nakayama(1,4), Hirohide Takebayashi(5), Junko Matsuda(2), Yasushi Enokido(1)
1. Institute for Developmental Research, Aichi Developmental Disability Center, 2. Dept. of Pathophysiology and Metabolism, Kawasaki Med. Sch., 3. Grad. Sch. of Med. Sci., Nagoya City Univ., 4. Grad. Sch. of Med., Nagoya Univ., 5. Grad. Sch. of Med. and Dent. Sci., Niigata Univ.
Keyword: microRNA, demyelination, myelin, lysosomal storage disease
Krabbe disease (Globoid cell leukodystrophy) is an autosomal recessive leukodystrophy manifested in an early developmental stage. Most of the patients are characterized by early-onset cerebral demyelination with apoptotic oligodendrocyte death and die before 2 years of age. It has been shown that galactosylceramidase (GALC) deficiency causes accumulation of cytotoxic sphingolipid, psychosine in oligodendrocyte. However, the mechanisms of Krabbe disease pathogenesis remain to be elucidated. We have demonstrated that primary cultured oligodendrocytes of twitcher mouse, an authentic mouse model of Krabbe disease, show abnormal oligodendrocyte development and accumulation of endogenous psychosine (Inamura et al., Neurobiol Dis 2018). In present study we show therapeutic efficacy of micro-RNA in twitcher oligodendrocytes. Expression of miR-219, which is critical for oligodendrocyte differentiation and maturation, was reduced in twitcher mouse oligodendrocytes in vivo and in vitro. Overexpression of miR-219 rescued oligodendrocyte development impaired in twitcher oligodendrocytes and reduced accumulation of psychosine and apoptotic death. Moreover, miR-219 reduced the activity of N-acylsphingosine amidohydrolase-1 (ASAH1), recently reported to generate psychosine, in twitcher oligodendrocytes. These results suggest that the reduction of miR-219 causes abnormal oligodendrocyte development in twitcher mouse and miR-219 supplementation has therapeutic potential for treatment of Krabbe disease oligodendrocyte pathologies. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-255
TSC2変異患者iPS細胞由来小脳オルガノイドの解析
Analysis of cerebellar organoids derived from patients carrying TSC2 mutation.
*奥村 啓樹(1,2)、有岡 祐子(1,3)、林 優(1)、久島 周(1)、森 大輔(1)、尾崎 紀夫(1)
1. 名古屋大学医学系研究科精神医学分野、2. 名古屋大学医学部附属病院薬剤部、3. 名古屋大学医学部附属病院先端医療開発部
*Hiroki Okumura(1,2), Yuko Arioka(1,3), Yu Hayashi(1), Itaru Kushima(1), Daisuke Mori(1), Norio Ozaki(1)
1. Dept of Psychiatry, Grad Sch Med, Nagoya Univ , 2. Dept of Hospital Pharmacy, Nagoya Univ Hospital, 3. CAMCR, Nagoya Univ Hospital
Keyword: iPS cells, cerebellar organoid, TSC2, autism spectrum disorder
Recent studies have revealed that the cerebellum is involved in various functions such as autonomic nervous system functions cognitive and emotional functions in conjunction with the cerebral cortex, not only regulation of motor function. Furthermore, the cerebellum has been implicated in various mental disorders, including autism spectrum disorder (ASD). In particular, Tuberous sclerosis complex (TSC), caused by abnormalities in the mutation of either TSC1 or TSC2, is a genetic disorder with high rates of comorbid ASDs. It has been reported that the cerebellar volume is smaller in TSC patients. Previous reports using the mouse model of TSC have shown that Purkinje cell dysfunction can cause ASD-related symptoms. However, it has been reported that brain structures and molecular mechanisms differ in mice from in humans. Whether they accurately recapitulate the phenotype of human diseases cannot be guaranteed. Thus the human experimental model system is required. Recently, it has been reported that the differentiation into cerebellar organoids from human ES/iPS cells by 3D self-organizing culture technologies. The cerebellar organoids contain various cells that constitute the cerebellum, such as Purkinje cells and granule cells, and may be useful as an in vitro human disease model. Therefore, we generated cerebellar organoids from TSC patient-derived iPS cells and analyzed their phenotype in this study. We generated iPS cells derived from three healthy Japanese subjects (HC) and three TSC patients with TSC2 mutation (TSC2-Pt). As previously reported, the cerebellar organoids were induced and analyzed by size measurement, qPCR, and immunostaining. The gene expression levels of Purkinje cells markers (KIRREL2, SKOR2, and CALB1), granule cell markers (BARHL1 and ATOH1), and Goldi cell marker (neurogranin) were time-dependently elevated in the cerebellar organoids. Furthermore, immunofluorescence analysis confirmed that these cerebellar organoids consist of cells expressing neuron marker (βIII-tubulin), neural stem cells marker (SOX2), and Purkinje cells markers (KIRREL2, SKOR2). However, the size of TSC2-Pt cerebellar organoids was significantly smaller than the HC cerebellum organoids in all cases. This result may be consistent with the small cerebellar volume in TSC patients. The TSC Patients derived cerebellar organoids may be useful for the human experimental model reflecting the cerebellar pathology in TSC patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-256
MECP2遺伝子欠損Rett症候群モデルマーモセットにおける全脳コネクトーム解析
Whole brain neural connectivity in the Rett syndrome model marmoset with MECP2 gene deletion
*吉丸 大輔(1,2,3,4)、岸 憲幸(2,4)、畑 純一(2,1,5)、釣木澤 朋和(3,2)、羽賀 柔(5,2)、岡野 ジェイムス洋尚(1)、岡野 栄之(2,4)
1. 東京慈恵会医科大学、2. 国立研究開発法人 理化学研究所 、3. 国立研究開発法人 産業技術総合研究所、4. 慶應義塾大学、5. 東京都立大学
*Daisuke Yoshimaru(1,2,3,4), Noriyuki Kishi(2,4), Junichi Hata(2,1,5), Tomokazu Tsurugizawa(3,2), Yawara Haga(5,2), Hirotaka James Okano(1), Hideyuki Okano(2,4)
1. The Jikei University School of Medicine, 2. RIKEN, 3. National Institute of Advanced Industrial Science and Technology (AIST), 4. Keio University , 5. Tokyo Metropolitan University
Keyword: DTI, Marmoset
Purpose: Rett syndrome (RTT) is an inherited neurological disorder caused by mutations in the methyl-CpG binding protein 2 (MECP2) gene. Characteristic brain findings of RTT include brain volume changes in human RTT patients and focal neural circuit abnormalities have been reported in MECP2 mutant mice. However, the characteristic neural connections of RTT in the whole brain are still unclear. In this study, we characterized the whole-brain neural connections of the MECP2 deletion common marmoset as a model for RTT. As a non-human primate, the marmoset has many similarities to humans in terms of sociality and central nervous system development, and we expect to gain important insights. Methods: In this study, MRI (Bruker BioSpin) was performed under isoflurane anesthesia in 1-3 month old MECP2 gene-deficient marmosets and wild-type marmosets. The whole brain was classified into 90 regions, and whole brain connectome analysis was performed from DTI data to evaluate the difference in neural connectivity between wild type and RTT in each brain region. For statistical analysis, we performed an unpaired t-test with Benjamin–Hochberg false discovery rate (FDR) correction (α=0.05), calculated the effect size using Cohen's d, and extracted and evaluated the more reliable neural connections as differences. Similarly, important features were extracted by least absolute shrinkage and selection operator (LASSO). Results: As the age of the child increased, the connections between brain regions in the RTT tended to decrease compared to the wild type. The connections with the largest effect sizes were between the medial prefrontal cortex (left) and middle temporal area (right). Based on feature extract by (LASSO), connections with small effect size, such as the thalamus (left) and the caudate nucleus (left), were also selected as important features. Conclusions: Whole brain neural connectivity analysis in the Rett syndrome model marmoset could characterize specific neural connectivity to RTT. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-257
ブメタミド慢性投与はアンジェルマン症候群モデルマウスの認知機能障害を改善する
Therapeutic effects of bumetanide on neurological dysfunction in a mouse model of Angelman syndrome
*江川 潔(1)、渡辺 美穂(2)、福田 敦夫(2)
1. 北海道大学医学部小児科、2. 浜松医科大学神経生理学講座
*Kiyoshi Egawa(1), Miho Watanabe(2), Fukuda Atsuo(2)
1. Pediatric dept, Hokkaido Univ. Grad. Med., 2. Dept. of Neurophysiology, Hamamatsu Univ. Sch. of Med.
Keyword: Angelman syndrome, Bumetanide, GABA
Angelman syndrome is a neurodevelopmental disorder caused by the loss of function of the maternally expressed gene UBE3A. The main clinical features consist of cognitive dysfunction, epilepsy, motor dysfunction, and behavioral abnormalities. Clinical symptoms overlap with those of autism spectrum disorders, including speech impairment and repetitive behavior. Recently, bumetanide, a loop diuretic, has been proposed as an effective compound for treating autism spectrum disorders by inhibiting the neuronal Na+-K+-Cl- cotransporter 1 (NKCC1), which facilitates Cl- influx. To investigate the therapeutic potential of bumetanide in Angelman syndrome, we analyzed Cl- hemostasis and the effects of bumetanide administration a mouse model of Angelman syndrome that lacks the maternal copy of the Ube3a gene (Ube3am–/p+). We found increased NKCC1 expression at the protein level in the hippocampus of Ube3am–/p+ mice. The steady-state intracellular Cl- concentration ([Cl-]i) of CA1 pyramidal neurons was not significantly different on average in these animals, but it demonstrated more variance in Ube3am–/p+ mice. As a possible mechanism counterbalancing the [Cl-]i increases due to NKCC1 activation, we demonstrated that tonic GABAA receptor-mediated Cl- influx was significantly reduced in CA1 pyramidal neurons. Chronic administration of bumetanide restored cognitive dysfunction in Ube3am–/p+ mice. Seizure susceptibility was also reduced by bumetanide in both Ube3am–/p+ and littermate control mice. These results suggest that [Cl-]i homeostasis is altered by multiple mechanisms, and aberrantly activated NKCC1 transporters have a pathophysiological impact leading to cognitive dysfunction in Ube3am–/p+ mice. Bumetanide administration might be effective for improving cognitive function and epilepsy in patients with Angelman syndrome. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-258
大脳発達期における神経成長関連蛋白質GAP43の役割
Role of GAP43 in the development of mouse cerebral cortex
*野田 万理子(1)、松本 歩(2)、伊東 秀記(1)、山形 崇倫(2)、永田 浩一(1)
1. 愛知県医療療育総合センター発達障害研究所、2. 自治医科大学医学部小児科学講座
*Mariko Noda(1), Ayumi Matsumoto(2), Hidenori Ito(1), Takanori Yamagata(2), Koh-ichi Nagata(1)
1. Inst Dev Res, Aichi Dev Disability Cntr, Aichi, Japan , 2. Dept Pediatrics, Jichi medical university
Keyword: GAP43, Neurodevelopmental disorder, intellectual disability
GAP43 (growth associated protein 43) is known to be expressed predominantly in the central nervous system. GAP43 is known for a crucial component of the axon and presynaptic terminals. Therefore, GAP43 is required for the navigation of axonal growth cones and new synapse formation.
Novel heterozygous missense variation (c.436 G>A, p.Glu146Lys) in the GAP43 gene is found in a patient with intellectual disability (ID) and motor retardation. However, the precise role of GAP43 during corticogenesis remains poorly defined.
We here examined the role of novel Gap43 mutant in mouse brain development. As first, protein stability of this variant in primary neurons was examined by western blotting. Since this variation leaded lower expression of Gap43 protein, gene knockdown was used to mimic the disease conditions. Acute silencing of Gap43 by in utero electroporation had little effects on excitatory neuron positioning during corticogenesis, whereas dendritic arbor formation and synaptogenesis was severely impaired. Based on these results, Gap43 may be critical for the maturation of cortical excitatory neurons and its functional defects are likely to be related to pathophysiology of ID. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-259
Cdkl5キナーゼ活性欠損ノックインマウスのin vivo大脳皮質広域カルシウムイメージングによる、CDKL5の機能欠損に伴う大規模な皮質ネットワークの機能的結合性変化の同定
In vivo wide-field Ca(2+) imaging of the cortical spontaneous activity in the Cdkl5 kinase-dead knock-in mice reveals altered functional connectivity in large-scale cortical networks upon the loss-of-function of CDKL5
*野村 俊貴(1)、村上 知成(2)、斎藤 久美子(2,4)、大木 研一(2,3)、田中 輝幸(1)
1. 東京大学医学系研究科発達医科学教室、2. 東京大学医学系研究科統合生理学教室、3. 東京大学ニューロインテリジェンス国際研究機構、4. 沖縄科学技術大学院大学神経回路ユニット
*Toshiki Nomura(1), Tomonari Murakami(2), Kumiko Saitou(2,4), Kenichi Ohki(2,3), Teruyuki Tanaka(1)
1. Department of Developmental Medical Sciences, Graduate School of Medicine, The University of Tokyo, 2. Department of Physiology, Graduate School of Medicine, The University of Tokyo, 3. International Research Center for Neurointelligence (IRCN), The University of Tokyo, 4. Neural Circuit Unit, Okinawa Institute of Science and Technology Graduate University (OIST)
Keyword: CDKL5, In vivo wide-field Ca2+ imaging, kinase-dead KI mouse, functional connectivity
Pathogenic mutations in the Cyclin-dependent kinase-like 5 (CDKL5) gene cause a severe neurodevelopmental disorder accompanied by early-onset intractable epilepsies, i.e. CDKL5 deficiency disorder (CDD). Recent studies of CDD patients and animal models have revealed the behavioral (macroscopic) and molecular (microscopic) phenotypes caused by the loss-of-function (LOF) of CDKL5. However, corresponding cortical circuits and connectivity alteration remain unclear, and identifying mesoscopic network-level functional connectivity (FC) alteration is necessary to understand the relationship between the molecular and behavioral phenotypes of CDD. We have recently generated the Cdkl5 K42R kinase-dead knock-in (KI) mice, in which the CDKL5 kinase activity is abolished. In order to determine how the loss of CDKL5 activity has an impact on the cortical neurocircuits, we have adopted in vivo wide-field Ca2+ imaging of the cortical spontaneous activity in the adult Cdkl5 kinase-dead KI and control mice in which GCaMP6f was neuronally expressed by the Thy1 promoter, and analyzed temporal correlation of the Ca2+ signals from entire cortices. The imaging was performed under anesthesia. We set 17 regions of interest (ROIs) in each hemicortex and analyzed the resting-state FC in brain cortices using a standard seed-based correlation method. We then evaluated differences in the intra- and inter-hemispheric FC across two genotypes by Student’s t-test. Cdkl5 kinase-dead KI mice showed significantly decreased intra- and inter-hemispheric FC compared to control mice. This result suggests that the spontaneous activity propagating through the networks in the cerebral cortex is impaired upon the LOF of CDKL5. We also compared FC in each ROI across two groups to identify more detailed network-specific alteration. This comparison indicated that FC between some cortical areas, such as motor, somatosensory and parietal areas were particularly decreased in Cdkl5 kinase-dead KI mice compared to control, whereas FC inside these areas were not strongly altered. It suggests that the FC in large-scale cortical networks rather than local neurocircuits is impaired, and some dominant brain networks like DMN may be functionally altered in CDD.
Since wide-field Ca2+ imaging is not readily applicable to observe the subcortical regions, resting-state fMRI analysis of Cdkl5 kinase-dead KI mice is ongoing in order to evaluate FC in the whole brain. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-260
神経発達症モデルマウスにおける眼球運動異常のメカニズムの解明
Mechanisms of eye movement abnormalities in mouse models of neurodevelopmental disorders
*入江 浩一郎(1,2)、臼井 紀好(1,3,4)、原田 祥太郎(5)、今井 貴夫(5)、島田 昌一(1,3,4)
1. 阪大・院医・神経細胞生物学、2. 阪大・院医・共同研、3. 阪大・院連合小児、4. 大阪精神医療セ・こころの科学・依存症、5. 阪大・院医・耳鼻咽喉科・頭頸部外科学
*Koichiro Irie(1,2), Noriyoshi Usui(1,3,4), Shotaro Harada(5), Takao Imai(5), Shoichi Shimada(1,3,4)
1. Dept. Neurosci. Cell Biol., Grad. Sch. Med., Osaka Univ., Osaka, Japan. , 2. CentMeRE., Grad. Sch. Med., Osaka Univ., Osaka, Japan., 3. UGSCD., Osaka Univ., Osaka, Japan., 4. Addiction., OPRC., OPMC., Osaka, Japan., 5. Dept. Otolaryngol. Head Neck Surg., Grad. Sch. Med., Osaka Univ., Osaka, Japan.
Keyword: Autism spectrum disorder, Neurodevelopmental disorder, Eye movement
Autism spectrum disorder (ASD) is a neurodevelopmental disorder exhibited deficits in social communication and limited repetitive behavior/interest. Abnormal eye movements have been previously observed in patients with neurodevelopmental disorders including ASD and attention-deficit/hyperactivity disorder. These abnormalities are expected to be used as a novel objective diagnostic method. However, the biological mechanisms underlying the eye movement abnormalities in patients with neurodevelopmental disorders remain largely unknown. In this study, to elucidate the mechanisms how eye movement abnormalities occur in patients with neurodevelopmental disorders. To establish a novel diagnostic method with eye movement, we evaluated the eye movement abnormalities using ASD mice models, and observed a decrease in the number of eye movements in ASD mice models. Furthermore, we performed histological analysis of ASD model mice brains. As a result, the number of mature neurons is decreased in the eye movement-related brain regions of ASD mice models. Our study suggests that the measurement of eye movement can be a potential biomarker in neurodevelopmental disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-261
フェンサイクリジンを亜慢性投与されたラットにおけるセロトニン5-HT2A受容体イメージング
Imaging of 5-HT2A receptors in rats with semi-chronic administration of phencyclidine
*浄土 英一(1)、中園 智晶(1)、高橋 和巳(1)、趙 松吉(2)、右近 直之(2)、鈴木 喜明(1)、星野 研洋(3)、永福 智志(1)
1. 福島県立医科大学医学部、2. 福島県立医科大学先端臨床研究センター、3. 一陽会病院
*Eiichi Jodo(1), Tomoaki Nakazono(1), Kazumi Takahashi(1), Songji Zhao (2), Naoyuki Ukon(2), Yoshiaki Suzuki(1), Kenyo Hoshino(3), Satoshi Eifuku(1)
1. Fukushima Medical University School of Medicine, Fukushima, Japan, 2. Advanced clinical research center, Fukushima Medical University, Fukushima, Japan, 3. Ichiyokai Hospital, Fukushima, Japan
Keyword: phencyclidine, schizophrenia, 18F-altanserin, 5-HT2A receptors
Phencyclidine (PCP) is a psychotomimetic drug that induces schizophrenia-like symptoms in healthy individuals and exacerbates pre-existing symptoms in patients with schizophrenia. PCP-treated animals are now considered a reliable pharmacological model of schizophrenia. We have shown that PCP induces tonic excitation of prefrontal cortex (PFC) neurons due to glutamatergic inputs from regions outside the PFC, and that this tonic activation of PFC neurons is a critical factor for the development of PCP-induced abnormal behaviors. Because glutamatergic transmission is partly modulated by serotonin via 5-HT2A receptors, PCP-treatment may affect 5-HT2A receptors. In this study we examined effects of semi-chronic administration of PCP (10 mg/kg/day, i.p., 15 times) on the binding potential (BP) of 5-HT2A receptors in rats using PET (positron-emission tomography) with 5-HT2A receptors-selective radioactive ligand, 18F-altanserin. Rats were divided into two groups, the PCP-treated group and the saline-treated one. Behavioral tests (sociability, social interaction) were executed twice (before the first drug administration and 2 days after the last drug treatment). Following the behavioral tests, PET signals were dynamically measured for 70 min on the same days as those of behavioral tests under volatile anesthesia (isoflurane). The radio ligand (25 MBq/0.4cc) was injected into the tail vein. The statistical analyses of behavioral tests data revealed that total travel distance was significantly longer in PCP-treated rats than the saline-treated ones in the sociability test, and that in the social interaction test the social interaction time between drug-treated- and unfamiliar rats was significantly shorter in PCP-treated rats. Since PET data are now being analyzed, we will present those data (BP, correlation between BP and behavioral data) at the meeting. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-262
リーリンモデルマウスにスパインの形態変化について研究
Spine morphological changes in Reln-del mice
*朱 悠韻(1)、常浦 祐未(2)、溝口 博之(1)、澤幡 雅仁(1)、森 大輔(1)、河野 孝夫(3)、服部 光治(3)、鍋島 俊隆(4)、尾崎 紀夫(1)、山田 清文(1)
1. 名古屋大学、2. 愛知県医療療育総合センター、3. 名古屋市立大学、4. 藤田医科大学
*YOUYUN ZHU(1), Yumi Tsuneura(2), Hiroyuki Mizoguchi(1), Masahito Sawahata(1), Daisuke Mori(1), Takao Kohno(3), Mitsuharu Hattori(3), Toshitaka Nabeshima(4), Norio Ozaki(1), Kiyofumi Yamada(1)
1. Nagoya University, 2. Aichi Developmental Disability Center, 3. Nagoya City University, 4. Fujita Health University
Keyword: Reelin, Spine, Schizophrenia, Cognition
Reelin is an extracellular matrix glycoprotein secreted by Cajal-Retzius cells and plays a critical role in neuronal migration and layer formation during brain development. In humans, deficiency of Reelin causes brain malformation, leading to a number of psychiatric diseases, such as schizophrenia (SCZ), bipolar disorder (BP) and autistic spectrum disorder. Schizophrenia includes deficits in cognition mediated by the circuitry of the dorsolateral prefrontal cortex (DLPFC) in humans, the region which has a similar function in rodents is the medial prefrontal cortex (mPFC). These deficits cause molecular and morphological changes in DLPFC, possibly influencing the neuroplasticity of the brain. Recently, we have developed transgenic mice carrying the same deletion in RELN encoding Reelin (Reln-del) which was identified in a Japanese schizophrenia patient. Reln-del mice showed abnormalities in their brain structure and social behavior. However, the details of dysfunction in the brain are unknown. Here, we performed an in vitro analysis of neuronal morphology of primary cultured cortical neurons and in vivo spine morphology analysis in wild-type (WT) and Reln-del mice. In vitro primary culture, Reln-del mice showed a significant decrease in both neurite length and branch point as well as in the number of postsynaptic density protein 95 (PSD95) immunoreactive puncta compared with WT mice. In vivo, the dendrite spine density was significantly decreased in the layers Ⅱ/Ⅲ and another of mPFC in Reln-del mice. Compared with WT mice, the spine of Reln-del mice showed a significant reduction of the stubby, mushroom and filopodia spines. Our results have identified that the exonic deletion of RELN plays an important role in the pathophysiology of SCZ. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-263
Reduced operant task engagement and altered neural oscillations in a triple-hit rat model of schizophrenia.
*Michele Chan(1), Qun Li(2), Gyongyi Horvath(2), Antal Berenyi(2,3,4), Masabumi Minami(1), Yuichi Takeuchi(1,2)
1. Dept Biopharmaceutical Sci Pharmacy, Facult Pharmaceutical Sci, Hokkaido Univ, Sapporo, Japan, 2. Dept Physiol, Facult Med, Univ Szeged, Szeged, Hungary, 3. HCEMM-SZTE Magnetotherapeutics Research Group, Univ Szeged, Szeged, Hungary, 4. Neurosci Inst, New York Univ, New York, USA
Keyword: Schizophrenia, Decision-Making, Neural Mechanism
Schizophrenia is a multi-dimensional neuropsychiatric disorder that affects decision-making, reward-taking behaviors and motivation. The subcortical limbic system together with the prefrontal cortex has been suggested to play a role in decision-making and reward-taking behaviors in schizophrenic patients. To examine the neural underpinnings of decision-making behavior in schizophrenia, a triple-hit schizophrenia model rat strain (WISKET) was subjected to operant learning tasks of increasing difficulty and a novel auditory go/no-go discrimination task. Local field potentials (LFPs) in the prefrontal cortices and the limbic system were recorded both during the one hour-long homecage session before the task and the go/no-go discrimination task. Behaviorally, we found that WISKETs performed similarly to the control Wistar rats in simple nose-poke tasks. However, when timing contingencies were added to the task, the task performance of WISKETs was significantly impaired compared to that of controls. Furthermore, performance of WISKETs in the go/no-go discrimination task was significantly impaired, showing lower levels of active trial initiation and reward-taking behavior compared to that of controls. This behavior is evident across footshock intensities ranged 0.2 to 0.5 mA. Power spectral analysis of LFP recordings in the homecage revealed that WISKETs had significantly increased LFP powers in the delta, theta, and beta frequency bands in the anterior-medial prefrontal cortex, ventral pallidum and ventral-hippocampus compared to those of controls. Although the increased LFP powers in the anterior-medial prefrontal cortex, ventral pallidum and ventral-hippocampus were also observed during the go/no-go task, the delta to gamma LFP powers in the ventral tegmental area were significantly lower in WISKET rats during the task. These findings suggest a neural basis of cognitive impairment in schizophrenia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-264
統合失調症の脳内意味ネットワーク異常に関するfMRI研究
Randomization of semantic network in the brain of schizophrenia revealed by fMRI
*松本 有紀子(1)、西田 知史(3)、林 隆介(5)、孫 樹洛(2)、村上 晶郎(2)、吉川 長伸(4)、伊藤 寛祥(6)、大石 直哉(2)、増田 直紀(7)、村井 俊哉(2)、Karl Friston(8)、西本 伸志(4)、高橋 英彦(1)
1. 東京医科歯科大学、2. 京都大学、3. 情報通信研究機構、4. 大阪大学、5. 産業技術総合研究所、6. 筑波大学
*Yukiko Matsumoto(1), Satoshi Nishida(3), Hayashi Ryusuke(5), Shuraku Son(2), Akio Murakami(2), Naganobu Yoshikawa(4), Hiroyoshi Ito(6), Naoya Oishi(2), Naoki Masuda(7), Toshiya Murai(2), Karl Friston(8), Shinji Nishimoto(4), Hidehiko Takahashi(1)
1. Tokyo Medical and Dental University, Tokyo, Japan, 2. Kyoto University, Kyoto, Japan, 3. National Institute of Information and Communications Technology (NICT), Osaka, Japan, 4. Osaka University, Osaka, Japan, 5. National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki, Japan, 6. University of Tsukuba, Ibaraki, Japan, 7. University at Buffalo, State University of New York, USA, 8. UCL Queen Square Institute of Neurology, London, UK
Keyword: Schizophrenia, fMRI, Semantic network, Small-worldness
Schizophrenia is a mental illness that presents with thought disorders including delusions and disorganized speech structures. Thought disorders have been regarded as a consequence of the loosening of associations between semantic concepts since the term “schizophrenia” was first coined by Bleuler in the early 20th century. However, a mechanistic account of this cardinal disturbance in terms of functional dysconnection has been lacking. To evaluate how aberrant semantic connections are expressed through brain activity, we characterized large-scale network structures of concept representations using functional magnetic resonance imaging. We quantified various concept representations in patients’ brains from functional magnetic resonance images of brain activity evoked by movie scenes using encoding modeling. We then constructed semantic brain networks by evaluating the similarity of these semantic representations and conducted graph theory-based network analyses. Neurotypical networks had small-world properties similar to those of natural languages, suggesting small-worldness as a universal property in semantic knowledge networks. Conversely, small-worldness was significantly reduced in networks of schizophrenia patients and was correlated with psychological measures of delusions. Patients’ semantic networks were partitioned into more-distinct categories and had more-random within-category structures than those of controls. These differences in conceptual representations manifest altered semantic clustering and associative intrusions that underlie thought disorders. This is the first study to provide pathophysiological evidence for the loosening of associations as reflected in randomization of semantic networks in schizophrenia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-265
刺激間間隔が聴性定常反応の同期活動に与える影響
Effect of duration of inter-stimulus-interval on phase-locking value of auditory steady-state responses
*鴻池 菜保(1)、岩沖 晴彦(1)、三輪 美樹(1)、伊藤 浩介(2)、中村 克樹(1)
1. 京都大学霊長類研究所、2. 新潟大学脳研究所
*Naho Konoike(1), Haruhiko Iwaoki(1), Miki Miwa(1), Kosuke Itoh(2), Katsuki Nakamura(1)
1. Primate Research Institute, Kyoto University, 2. Brain Research Institute, University of Niigata
Keyword: schizophrenia, ASSR, EEG, common marmosets
Auditory steady-state responses (ASSRs) are basic neural responses used to probe the ability of auditory circuits to produce synchronous activity to repetitive external stimulation. Reduced ASSR in power and/or phase-locking activity has been observed in patients with schizophrenia, especially at 40 Hz. Therefore, ASSR can be a useful translational biomarker for schizophrenia and other psychiatric disorders. In the present study, we examined if the duration of inter-stimulus-interval (ISI) would affect the phase-locking value of ASSRs. Five young-adult healthy Japanese volunteers (1 female and 4 males, 24-27 years old) participated in the experiment. We used 1-ms click trains at 30, 40, or 80 Hz as auditory stimuli. The train duration was 500 ms, and the ISI was chosen from the possible five intervals (400, 600, 800, 1000, or 1200 ms) for each session. We found that the longer ISIs elicited the larger phase-locking values at 40 Hz-ASSR. Recently, common marmosets are thought to be useful as animal models for neurological and psychiatric disorders. However, it is still unclear whether or not the same ASSR paradigm as for humans is appropriate for evaluating animal models of schizophrenia. In the current study, we attempted to compare the brain response to auditory repetitive stimuli between humans and marmosets by electroencephalography (EEG) with a non-invasive method. Five adult common marmosets (Callithrix jacchus, 2 female and 3 male, 5-8 years old, 272-342g) participated in the experiments. The recording methods have been previously described for the marmoset (Itoh et al., 2021). We found that the duration of ISI changed the phase-locking value at 40 Hz-ASSR also in the marmosets. These results may provide us with useful information on appropriate parameters to evaluate ASSRs in humans and marmosets. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-266
樹状突起スパインの樹状突起区画特異的蓄積は、青年期における大脳皮質回路成熟の特徴である。
Dendritic compartment-specific accumulation of dendritic spines is a hallmark of cortical circuit maturation during adolescence
*江頭 諒(1)、柯 孟岑(2)、高木 豪(3,4)、石井 俊輔(4)、宮川 剛(5)、今井 猛(1)
1. 九州大学大学院 医学研究院、2. 理化学研究所 多細胞システム形成研究センター、3. 愛知県医療療育総合センター発達障害研究所、4. 理化学研究所 筑波地区、5. 藤田医科大学 総合医科学研究所 システム医科学研究部門
*Ryo Egashira(1), Meng-Tsen Ke(2), Tsuyoshi Takagi(3,4), Shunsuke Isii(4), Tsuyoshi Miyakawa(5), Takeshi Imai(1)
1. Kyushu University Graduate School of Medical Sciences, 2. RIKEN Center for Developmental Biology, Kobe, Japan, 3. Institute for Developmental Research, Aichi Human Service Center, Kasugai, Japan, 4. RIKEN Tsukuba Institute, Tsukuba, Japan, 5. Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan
Keyword: Schizophrenia, Adolescence, Spine density hotspot, Layer 5 pyramidal tract neuron
Our brain acquires higher cognitive functions during adolescence. It has been generally assumed that the number of synapses in the cerebral cortex increases during childhood but decreases during adolescence. As synaptic density is reduced in schizophrenia patients, it has been hypothesized that over-pruning of synapses is a cause of this neuropsychiatric disorder. In our previous study, however, we found that spine density increases during adolescence at specific compartments in apical dendrites of layer5 pyramidal tract (L5 PT) neurons in mice. Moreover, we obtained no clear evidence for spine density reduction in other compartments of dendrites in cortical pyramidal neurons during adolescence. These results suggest that compartment-specific spine accumulation is a major change of the cerebral cortex during adolescence. In this study, we tested a hypothesis in which defects in the compartment-specific accumulation of dendritic spines, rather than the pruning, are associated with schizophrenia that develops during adolescence. We examined Schnurri-2 (Shn2) deficient mice which are known to show various abnormal behaviors related to schizophrenia. At P14, the spine density in Shn2 knockout was similar to that of controls in both apical and basal dendrites of L5 PT neurons. In adult mice, however, the spine density was preferentially reduced in apical dendrites in the knockouts compared to the controls. We also examined the impact of Grin1 knockout in synapse development in L5 PT neurons. Grin1 encodes the essential subunits of the NMDA receptors, NR1. AAV carrying Cre was introduced to Grin1 floxed mouse at P14 and spine density was analyzed in the adults. In this case, spine density was reduced specifically apical dendrites compared to the controls. Our study suggests that regulation of spine accumulation, rather than pruning, is important for the normal development of cognitive functions during adolescence. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-267
神経発火異常を引き起こす巨大スパインの非線形コンピュテーション
Distorted neurocomputation by heavily-weighted synapses in mental disorders
*小尾（永田） 紀翔(1)、鈴木 紀光(1)、Matthew L. MacDonald(4)、Kenneth N. Fish(4)、白井 福寿(1)、沖村 宰(2)、長濱 健一郎(5,6)、田中 昌司(3)、狩野 方伸(5,6)、Robert A. Sweet(4)、林（高木） 朗子(1)
1. 理化学研究所・脳神経科学研究センター、2. 慶応義塾大学医学部、3. 上智大学理工学部、4. ピッツバーグ大学、5. 東京大学大学院医学系研究科、6. 東京大学国際高等研究所ニューロインテリジェンス国際研究機構
*Kisho Obi-Nagata(1), Norimitsu Suzuki(1), Matthew L. MacDonald(4), Kenneth N. Fish(4), Fukutoshi Shirai(1), Tsukasa Okimura(2), Kenichiro Nagahama(5,6), Shoji Tanaka(3), Masanobu Kano(5,6), Robert A. Sweet(4), Akiko Hayashi-Takagi(1)
1. RIKEN Center for Brain Science, 2. Keio University, Tokyo, Japan, 3. Sophia University, Tokyo, Japan, 4. University of Pittsburgh, Pennsylvania, US, 5. Graduate School of Medicine, The University of Tokyo, Tokyo, Japan, 6. International Research Center for Neurointelligence (WPI-IRCN), The University of Tokyo, Tokyo, Japan
Keyword: Dendritic spine, Dendritic computation, Prefrontal cortex, Postmortem brain
The spatiotemporal organization of neuronal firing is crucial for information processing, and how thousands of synaptic inputs to the dendritic spines drive the firing remains a central question in neuroscience. Despite the accumulating evidence that has suggested a change in synaptic density and strength as a possible pathophysiology of various psychiatric disorders, it is unknown whether the synaptopathy an underlying pathogenesis of the disorders or a secondary consequence. In other words, it is entirely unclear what changes in neuronal computation and subsequent dynamism of neuronal circuits would be evoked when such synaptic change occurs. To address this question, we performed multi-scale synapse analyses, in which dendrite and somatic events are simultaneously assessed during precise stimulation of identified spines by two-photon glutamate uncaging. Interestingly, mice with knockdown of SETD1A and DISC1, both well-established animal models for schizophrenia, exhibited a significant number of extra-large (XL) spines (corresponding to more than 2.5 standard deviations from the average of normal mice). We found XL spines evoke a marked supralinear synaptic amplification, and clustered inputs to a few XL spines were sufficient to drive neuronal firing. We experimentally and theoretically observed that the cluster density of XL spines negatively correlated with working memory, which can contribute to psychiatric pathophysiology. Furthermore, the cluster density of XL spines was significantly overrepresented in the postmortem brain of schizophrenia than age- and gender-matched control. The currently dominant hypothesis of schizophrenia pathophysiology is a reduction in spine density (the over-pruning hypothesis). However, our results presented here suggest that the hypothesis may need to be revised and that more in-depth investigations into the interactions between intra-spine and dendritic computations and their effect on brain (dys)function are needed. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-268
統合失調症マウスモデルにおける臨床を反映した抗精神病薬投与下での認知機能障害の新規治療戦略
A novel therapeutic strategy for cognitive dysfunctions under clinically relevant antipsychotic treatment in a mouse model of schizophrenia
*有銘 預世布(1)、斎藤 喜人(1)、内田 康雄(2)、石河 三貴子(1)、上吉原 千賀子(1)、藤井 一希(3,4)、高雄 啓三(3,4)、大川 宜昭(1)
1. 獨協医科大学先端医科学研究センター、2. 東北大学大学院薬学研究科、3. 富山大学学術研究部医学系、4. 富山大学研究推進総合支援センター
*Yosefu Arime(1), Yoshito Saitoh(1), Yasuo Uchida(2), Mikiko Ishikawa(1), Chikako Kamiyoshihara(1), Kazuki Fujii(3,4), Keizo Takao(3,4), Noriaki Ohkawa(1)
1. Res Cen Adv Med Sci, Dokkyo Med Univ, Mibu, Japan, 2. Grad Sch Pharm Sci, Univ of Tohoku, Sendai, Japan, 3. Fac Med, Univ of Toyama, Toyama, Japan, 4. Life Sci Res Cen, Univ of Toyama, Toyama, Japan
Keyword: WORKING MEMORY, PRELIMBIC CORTEX, PARVALBUMIN, CHEMOGENETICS
Background:Deficits in cognitive functions (e.g., working memory, executive functions) are core features of schizophrenia. These are more predictive for functional outcomes of schizophrenia than positive and negative symptoms, however, there are currently no approved medications. Continuous antipsychotic medication is indispensable for not only improving positive symptoms in acute phase but also relapse prevention in maintenance phase. To develop novel drug therapies to cover both of cognitive deficits and positive symptoms, it is necessary to understand the neural mechanisms closely linking with the features under clinically comparable antipsychotic treatment. However, candidate mechanisms to improve the deficits are poorly understood. Methods:Here, we used chronic phencyclidine (PCP)-treated mice as an animal model of schizophrenia and tried multi-dimensional methods, including measurement of LC-MS/MS-based in vivo D2 receptor occupancy, histological analysis of prefrontal dendritic spines and synapses, and behavioral analyses of mice with chemogenetic manipulation, to investigate neural mechanisms and treatment strategies for working memory deficits in this mouse model. Results:In the layer 2–3 of the prelimbic cortex of chronic PCP-treated mice, we found that the number of dendritic spines, VGLUT1+ puncta and parvalbumin (PV) + puncta onto the pyramidal neuron soma were decreased. Continuous olanzapine dose regimen to PCP-treated mice, which achieve a sustained therapeutic window of dopamine D2 receptor occupancy (65–80%) in the mouse striatum, partially improved VGLUT1+ synapse abnormalities, but did not ameliorate PV+ synapse and working memory impairments. These results raise a possibility that abnormal excitatory/inhibitory balance in this area is linking with cognitive deficits under clinical conditions of schizophrenia. To improve the balance, we tried selective activation of parvalbumin interneurons in the prelimbic cortex, using hM3D(Gq) DREADD and its selective ligand descloroclozapine. By this modulation, the working memory deficits in chronic PCP-treated mice were restored. Furthermore, this improvement effect was maintained even under continuous olanzapine treatment. Discussion:These results suggest that selective activation of prefrontal parvalbumin interneurons in combination with clinically comparable olanzapine treatment provides more translational approaches to facilitate novel drug discovery improving cognitive deficits in schizophrenia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-269
統合失調症における病理学的神経回路の形成メカニズム解析
Analysis of the mechanism of pathological neural circuit formation in schizophrenia mouse model
*福田 茉由(1)、向井 淳(1)
1. 筑波大学 プレシジョン・メディスン開発研究センター
*MAYU FUKUDA(1), JUN MUKAI(1)
1. PMC, Univ of Tsukuba, Tokyo, Japan
Keyword: Schizophrenia, WORKING MEMORY, Setd1a, Precision Psychiatry
Schizophrenia (SCZ) is a psychiatric disorder that affects about 1% of the population in any region of the world. SCZ has positive symptoms (hallucinations and delusions, etc.), negative symptoms (social withdrawal and lethargy, etc.), and cognitive impairment (reduced memory, decreased attention and decreased information processing ability). Cognitive dysfunction is a key element of schizophrenia and is resistant to long-term treatment, impeding the patient's social life and rehabilitation. Working memory, which forms the core of cognitive function, is a fundamental brain process used to retain and recall information on the fly. When working memory impairment occurs, it has a serious impact on reasoning, cognition, decision making, and so on.
Currently, there are no approved therapeutic agents for cognitive dysfunction, and the development of innovative therapeutic agents and treatment methods is required. The fact that long-term morbidity increases in the absence of any treatment strongly reinforces the need for early interventions before pathological defects cascade into an irreversible state. The purpose of this study is to elucidate the mechanism of pathological neural circuit formation related to cognitive function in schizophrenia, identify the developmental windows where therapeutic intervention is possible, and develop new therapeutic methods. Using a monogenic mouse model (Setd1a+/-) with a definite causal relationship with schizophrenia, we performed various omics analyzes to relate the structure and function of genome with the disease. As a result, differential expression of genes associated with schizophrenia was observed in RNA-Seq of developing mice. Our goal is offer novel conceptual advances regarding the mechanism of pathological neural circuit formation of the developing brain in schizophrenia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-270
Importin α4/KPNA4 ノックアウトマウスの行動異常の背景にある分子基盤の探索
Investigation of the molecular background behind the behavioral deficits seen in Importin α4/KPNA4 Knockout Mice
*櫻井 航輝(1)、宮本 洋一(2)、森田 真規子(1)、マクファーソン トム(1)、小澤 貴明(1)、岡 正啓(2)、疋田 貴俊(1)
1. 大阪大学 蛋白質研究所、2. 医薬基盤研究所 核輸送ダイナミクスプロジェクト
*Koki Sakurai(1), Yoichi Miyamoto(2), Makiko Morita(1), Tom Macpherson(1), Takaaki Ozawa(1), Masahiro Oka(2), Takatoshi Hikida(1)
1. Institute for Protein Research, Osaka Univ., 2. Laboratory of Nuclear Transport Dynamics, National Institute for Biomedical Innovation
Keyword: Importinα, Monoamine Metabolism
Importins are a family of proteins that assist newly synthesized polypeptide molecules to be transported from the cytoplasm into the nucleus. Importin αs (Imp αs), otherwise known as Karyopherin αs (KPNAs), are a type of Importin that bind to classical nuclear localization signals contained in the sequences of cargo proteins and form a cargo, Impα, and Importin β trimer, which allow the cargo to move through nuclear pore complexes into the nucleus. The characteristics of Imp αs related to nucleocytoplasmic transport have been extensively studied, but the physiological implications of such cellular functions are still under investigation. Several of the 7 Impα subtypes known in humans (6 in mice) have been associated to psychiatric disease and/or alterations in mouse behavior, bringing up the importance of Impαs in brain function and behavioral regulation.
In particular, the Importin α4 protein (KPNA4) is known to transport NF-κB transcription factors, and its decrease has been associated to inflammatory signals and schizophrenia. The behavioral phenotypes of Kpna4 KO mice were examined by conducting a battery of behavioral tasks. A KO mice line lacking KPNA4 showed behavioral abnormalities including increased anxiety and reduced sensorimotor gating, providing evidence that KPNA4 is associated with brain function and behavior. To further characterize the molecular and biochemical states occurring in these KO mice, we performed neurochemical characterization. HPLC quantification of monoamine neurotransmitters and its metabolites in several brain regions showed alterations in monoamine neurotransmitter metabolite levels in Kpna4 KO mice. We aim to elucidate further details in the molecular background underlying the behavioral and neurochemical abnormalities through characterization of cellular gene expression. Further insight into the mechanisms may provide new insights on the brain functions and pathology for neuropsychiatric disorders. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-271
統合失調症患者由来神経系細胞の転写制御ネットワーク再構築およびメタアナリシス
Meta-analysis of transcriptional regulatory networks for lipid metabolism in neural cells from schizophrenia patients based on an open-source intelligence approach
*柚木 克之(1,2,3)、岡本 理沙(1,2)、渡部 素世香(1,2)、出野 泉花(1,2)、聶 翔(1)、丸山 順一(1)、冨田 勝(2)、幡野 敦(1,4)
1. 理化学研究所生命医科学研究センター、2. 慶應義塾大学先端生命科学研究所、3. 東京大学理学系研究科、4. 新潟大学医歯学系
*Katsuyuki Yugi(1,2,3), Lisa Okamoto(1,2), Soyoka Watanabe(1,2), Senka Deno(1,2), Xiang Nie(1), Junichi Maruyama(1), Masaru Tomita(2), Atsushi Hatano(1,4)
1. RIKEN IMS, Yokohama, Japan, 2. Inst Adv Biosci, Keio Univ, Fujisawa, Japan, 3. Grad Sch Sci, Univ of Tokyo, Tokyo, Japan, 4. Grad Sch Med Dent Sci, Niigata Univ, Niigata, Japan
Keyword: Transcriptional regulatory network, Enhancer, Lipid metabolism, Meta-analysis
There have been a number of reports about the transcriptional regulatory networks related to schizophrenia. However, most of these studies were based on a specific transcription factor or a single dataset, an approach that is not necessarily appropriate to understand the diverse etiology and underlying common characteristics of schizophrenia. Here we reconstructed and compared the transcriptional regulatory networks for lipid metabolism in brain using 15 public transcriptome datasets of neural cells obtained from schizophrenia patients. Since many of the well-known schizophrenia-related SNPs are in enhancers, we reconstructed a network including not only promoter-dependent regulation but also enhancer-dependent regulation, and found that 53.3 % of the total number of edges (7,577 pairs) involved regulation via enhancers. By examining multiple datasets, we found common and unique subnetworks that may constitute transcriptional modes of regulation. Furthermore, enrichment analysis of trait/disease annotations of SNPs that were connected with genes in the transcriptional regulatory networks by eQTL suggested an association with hematological cell counts, eye system, and some other traits/diseases whose relationship to schizophrenia was either not or insufficiently reported in previous studies. Based on these results, we suggest that in future studies on schizophrenia, information on genotype, comorbidities and hematological cell counts should be included, along with the transcriptome, for a more detailed genetic stratification and mechanistic exploration of schizophrenia. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-272
精神疾患の発症機序と新規遺伝子発現調節系「LDB2-EGR軸」の関連性
Roles of Lim-domain binding 2 as a functional modulator of EGR transcription factors and its implication in the pathogenesis of mental disorders
*大西 哲生(1,2)、光増 真広(1)、城山 優治(5,7)、有馬-吉田 史子(9,7)、門田 満隆(12)、岩山 佳美(10)、市川 智恵(11,3)、山田 一之(4)、中谷 明弘(13)、新井 誠(3)、堀内 泰江(3)、島本-光山 知恵(2)、前川 素子(2,8)、糸川 昌成(3)、尾藤 晴彦(6)、工樂 樹洋(12)、真鍋 俊也(7)、奥野 浩行(5)、吉川 武男(2)、田中 光一(1)
1. 東京医科歯科大学難治疾患研究所、2. 理化学研究所脳神経科学研究センター、3. 東京都医学総合研究所、4. 静岡産業大学、5. 鹿児島大学大学院医歯学総合研究科、6. 東京大学大学院医学系研究科、7. 東京大学医科学研究所、8. 東北大学大学院医学系研究科、9. 慈恵医大臨床医学研究所、10. 理化学研究所、11. 明治薬科大学、12. 理化学研究所生命機能科学研究センター、13. 東京大学大学院新領域創成科学研究科
*Tetsuo ohnishi(1,2), Masahiro Mitsumasu(1), Yuji Kiyama(5,7), Fumiko Arima-Yoshida(9,7), Mistutaka Kadota(12), Yoshimi Tanaka(10), Tomoe Ichikawa(11,3), Kazuyuki Yamada(4), Akihiro Nakaya(13), Makoto Arai(3), Yasue Horiuchi(3), Chie Shimamoto-Mitsuyama(2), Motoko Maekawa(2,8), Masanari Itokawa(3), Haruhiko Bito(6), Shigehiro Kuraku(12), Toshiya Manabe(7), Hiroyuki Okuno(5), Takeo Yoshikawa(2), Kohichi Tanaka(1)
1. Inst of Med Res, Tokyo Med Dent Univ, Tokyo, Japan, 2. RIKEN CBS, Wako, Japan, 3. Tokyo Metropl Inst of Med Sci, Tokyo, Japan, 4. Shizuoka Sangyo Univ, Iwata, Japan, 5. Grad Sch of Med Dent Sci, Kagoshima Univ, Kagoshima, Japan, 6. Grad Sch of Med, the Univ of Tokyo, Tokyo, Japan, 7. IMS, the Univ of Tokyo, Tokyo, Japan, 8. Tohoku Univ Sch of Med, Sendai, Japan, 9. Inst of Clin Med Res, Jikei Univ, Tokyo, Japan, 10. RIKEN, Wako, Japan, 11. Meiji Pharmaceut Univ, Kiyose, Japan, 12. RIKEN BDR, Kobe, Japan, 13. Grad Sch of Front Sci, the Univ of Tokyo, Tokyo, Japan
Keyword: schizophrenia, EGR, LDB2, balanced chromosomal translocation
To date, studies have identified numerous ‘risk’ variants and genes for mental disorders. However, most of those variants are common in the population, and, in many cases, the effect size of those variants is minimum. Exceptional genomic defects with large effect size, even though they are minor in the population, can help elucidate unknown pathologic architecture of mental disorders. We previously reported a patient with schizophrenia and a balanced translocation between chromosomes 4 and 13 and found that the breakpoint within chromosome 4 is located near the LDB2 gene, which encodes a putative transcription regulator. In this presentation, we show several lines of evidence suggesting that functional deficits of the gene evoked by the chromosomal break was implicated in the pathogenesis of the proband patient. First, expression of LDB2 was significantly lower in neurosphere cells differentiated from the patient’s iPS cells than in those from a control subject without psychotic symptoms or similar chromosomal translocation. Second, Ldb2 knockout (KO) mice displayed multiple deficits relevant to mental disorders. In particular, Ldb2 KO mice exhibited hyperactivity in various behavioral tasks and remarkable deficits in the fear-conditioning paradigm. We next explored the molecular mechanism in which Ldb2 KO mice displayed these schizophrenia-relevant behavioral alternations. Since the amygdala is known to play a pivotal role in executing the fear-conditioning paradigm, we focused on the amygdala. Analysis of the amygdala suggested that dysregulation of synaptic activities controlled by the immediate early gene Arc is involved in the phenotypes. We also show that LDB2, which lacks known DNA binding domains, forms protein complexes with known transcription factors. Consistently, expressions of > 300 genes were found to be dysregulated in the brain from Ldb2 KO mice. Furthermore, ChIP-seq analyses indicated that the LDB2 protein binds to > 10,000 genomic sites in human neurospheres differentiated from iPS cells. We found that many of those sites, including the promoter region of ARC, are occupied by EGR transcription factors. Our previous study showed an association of the EGR family genes with schizophrenia (Yamada et al., PNAS, 2007). Taken together, the findings suggest that dysregulation in the gene expression controlled by the LDB2-EGR axis underlies a pathogenesis of subset of mental disorders. Additional data on the molecular dissection of this gene expression system will be presented. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-273
精神疾患横断的AMPA受容体分布の検討
Cross-sectional AMPA Receptor Distribution in Psychiatric Disorders
*波多野 真依(1)、宮崎 智之(1)、中島 和希(1)、永露 毅(1)、中野 晃太郎(1)、木村 裕一(2)、内田 裕之(3)、高橋 琢哉(1)
1. 横浜市立大学医学研究科生理学、2. 近畿大学生物理工学部生命情報工学科、3. 慶応義塾大学医学部精神・神経科学教室
*Mai Hatano(1), Tomoyuki Miyazaki(1), Waki Nakajima(1), Tsuyoshi Eiro(1), Kotaro Nakano(1), Yuichi Kimura(2), Hiroyuki Uchida(3), Takuya Takahashi(1)
1. Department of Physiology Yokohama City University graduate school of medicine, 2. Faculty of Biology-Oriented Science and Technology, Kindai University, Kinokawa, Japan, 3. Department of Neuropsychiatry, Keio University School of Medicine, Tokyo, Japan
Keyword: AMPA receptor, Psychiatric disorder, Schizophrenia, PET
グルタミン酸シナプスは興奮性シナプスの約9割を占める。α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptor（AMPA）受容体はグルタミン酸受容体の一つであり、グルタミン酸シナプスの機能を中核的に担っている。精神疾患の原因の一つにAMPA受容体の発現量の変化や機能異常が考えられ、げっ歯類を用いた基礎研究や死後脳用いた研究が数多く行われてきた(Toth et al., 2008; Rubio et al., 2012)。しかし、これまで生体内のAMPA受容体を可視化した研究はなかった。そこで我々はAMPA受容体に結合するPET薬剤[11C]K-2を開発したた(Miyazaki et al.,l。、 2020)。 精神疾患は、、これまで、臨床症状による診断や、疾患の分類が行われてきた。しかし、現状の治療法では、治療薬が効かない難治例も多く、日常生活に著しい制限をもたらすが多く存在する問題点が挙げられる。その理由として、生物学的メカニズムが不明であること、また、、診断における対する客観的指標に乏しいことがないことが挙げられる。解決方策として、これまでの縦割りの疾患分類ではなく、生物学的背景に基づいた疾患横断的分類が重要であると考えた。本研究では、AMPA受容体標識PET薬剤[11C]K-2を用いて、精神疾患横断的な脳内AMPA受容体分布を明らかにすることを目的とした。 統合失調症、うつ病、双極性障害、自閉症スペクトラム障害、健常者の症例を[11C]K-2 PET、MRI (T1画像、resting state functional MRI (rs-fMRI))の撮像を行った。まず、健常者と精神疾患患者におけるAMPA受容体分布を比較した。健常者と比較して精神疾患群では、前頭葉、前部帯状回、前方の島皮質ではAMPA受容体量の低下を示し、後頭葉では増加を示した。これらの領域は精神疾患に共通して変化する責任領域と考え、trait領域と規定した。次に、統合失調症患者において臨床スコア（PANSS: positive and negative symptom scale）とAMPA受容体量の相関を解析した。前部帯状回膝前部膝下部、後部帯状回、島皮質、被殻において負の相関が認められ、これらの領域は統合失調症の症状を表す領域としてstate領域と規定した。最後に、精神疾患横断的trait領域とstate領域間の関係性を検討するためにrs-fMRI解析を行った。統合失調症患者と健常者における機能的結合性を比較したところ、Frontal trait(前頭葉や前部帯状回)、Lateral trait (島皮質)、Occipital trait (後頭葉)の領域間に機能的結合性の変化を認めた。また、Lateral traitとstate領域の機能的結合性が統合失調症群において低下していることが明らかになった。 以上の結果から、[11C]K-2は症状や状態を反映する客観的バイオマーカーになる得ること、また、生物学的背景に基づいた疾患横断的な分類の新たな可能性が広がることが示唆された。 | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-274
主要精神疾患および神経発達障害のポリジェニックリスクスコアは幼少時の睡眠障害に寄与する: Adolescent Brain Cognitive Development (ABCD) 研究
Polygenic risk scores for major psychiatric and neurodevelopmental disorders contribute to sleep disturbance in childhood: Adolescent Brain Cognitive Development (ABCD) Study
*大井 一高(1)、Ryo Ochi(2)、Yoshihiro Noda(2)、Masataka Wada(2)、Shunsuke Sugiyama(1)、Akira Nishi(2)、Toshiki Shioiri(1)、Masaru Mimura(2)、Shinichiro Nakajima(2)
1. 岐阜大学大学院医学系研究科精神医学、2. 慶應義塾大学医学部精神神経科学
*Kazutaka Ohi(1), Ryo Ochi(2), Yoshihiro Noda(2), Masataka Wada(2), Shunsuke Sugiyama(1), Akira Nishi(2), Toshiki Shioiri(1), Masaru Mimura(2), Shinichiro Nakajima(2)
1. Department of Psychiatry, Gifu University Graduate School of Medicine, 2. Department of Neuropsychiatry, Keio University School of Medicine
Keyword: Attention-deficit/hyperactivity disorder, major depressive disorder, polygenic risk score, sleep disturbance
Sleep disturbance is a common symptom of psychiatric and neurodevelopmental disorders and, especially in childhood, can be a precursor to various mental disorders. However, the genetic etiology of mental illness that contributes to sleep disturbance during childhood is poorly understood. We investigated whether the polygenic features of psychiatric and neurodevelopmental disorders are associated with sleep disturbance during childhood. We conducted polygenic risk score (PRS) analyses by utilizing large-scale genome-wide association studies (GWASs) (n = 46,350-500,199) of five major psychiatric and neurodevelopmental disorders (autism spectrum disorder, schizophrenia, attention-deficit/hyperactivity disorder (ADHD), major depressive disorder (MDD), and bipolar disorder) and, additionally, anxiety disorders as base datasets. We used the data of 9- to 10-year-olds from the Adolescent Brain Cognitive Development study (n = 9683) as a target dataset. Sleep disturbance was assessed based on the Sleep Disturbance Scale for Children (SDSC) scores. The effects of PRSs for these psychiatric and neurodevelopmental disorders on the total scores and six subscale scores of the SDSC were investigated. Of the PRSs for the five psychiatric and neurodevelopmental disorders, the PRSs for ADHD and MDD positively correlated with sleep disturbance in children (ADHD: R2 = 0.0033, p = 6.19 × 10-5, MDD: R2 = 0.0042, p = 5.69 × 10-6). Regarding the six subscale scores of the SDSC, the PRSs for ADHD positively correlated with both disorders of initiating and maintaining sleep (R2 = 0.0028, p = 2.31 × 10-4) and excessive somnolence (R2 = 0.0023, p = 8.44 × 10-4). Furthermore, the PRSs for MDD primarily positively correlated with disorders of initiating and maintaining sleep (R2 = 0.0048, p = 1.26 × 10-6), followed by excessive somnolence (R2 = 0.0023, p = 7.74 × 10-4) and sleep hyperhidrosis (R2 = 0.0014, p = 9.55 × 10-3). Despite high genetic overlap between MDD and anxiety disorders, PRSs for anxiety disorders correlated with different types of sleep disturbances such as disorders of arousal or nightmares (R2 = 0.0013, p = 0.011). These findings suggest that greater genetic susceptibility to specific psychiatric and neurodevelopmental disorders, as represented by ADHD, MDD, and anxiety disorders, may contribute to greater sleep problems among children. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-275
N-アセチル基転移酵素Shati/Nat8lによって制御される背側線条体-背側縫線核回路が線条体のセロトニン制御を介してストレス感受性を決定する
The dorsal striatum-dorsal raphe nucleus circuit regulated by N-acetyltransferase, Shati/Nat8l, determine the sensitivity to stress through striatal 5-HT control
*宮西 肇(1)、村松 慎一(2,3)、三枝 理博(4)、新田 淳美(1)
1. 富山大学大学院医学薬学教育部薬物治療学、2. 自治医科大学オープンイノベーションセンター神経遺伝子治療、3. 東京大学医科学研究所遺伝子・細胞治療センター、4. 金沢大学医薬保健研究域医学系統合神経生理学
*Hajime Miyanishi(1), Shin-ichi Muramatsu(2,3), Michihiro Mieda(4), Atsumi Nitta(1)
1. Dept Pharm Therap and Neuropharmacol, Grad Sch Med and Pharm, Univ of Toyama, 2. Div Neuro Gene Therap, Open Innov Ctr, Univ of Jichi Med, 3. Ctr Gene and Cell Therap, Ins Med Sci, Univ of Tokyo, 4. Dept Integr Neurophysiol, Grad Sch Med Sci, Univ of Kanazawa
Keyword: depression, stress , dorsal stritum, dorsal raphe nucleus
[Background] The number of patients with depression have been increasing in over the world. Stress is closely related to the onset of depression. However, chronic stress does not always trigger depression, as some individuals are resilient. Revealing the underlying regulatory mechanisms of stress sensitivity could offer novel insights for understanding the pathogenesis of depression. We found that Shati/Nat8l, N-acetyltransferase, levels increased in the dorsal striatum of stress-susceptible, not resilient, mice exposed repeated social defeat stress (RSDS). In the present study, we demonstrated that the regulation mechanism of stress sensitivity by Shati/Nat8l in the dorsal striatum. [Methods] C57BL/6J male mice were exposed RSDS using ICR mice, and the stress susceptible or resilient group were classified by social interaction test. We generated dorsal striatal Shati/Nat8l overexpression (dSTR-Shati OE) mice. Depression-like behaviors were assessed in these mice after RSDS or sub-threshold social stress (microdefeat stress), and 5-HT levels in the dorsal striatum were measured using in vivo microdialysis. The effects of 5-HT in the dorsal striatum to stress sensitivity regulated by striatal Shati/Nat8l were investigated by microinjection of selective serotonin reuptake inhibitors (SSRI) into the dorsal striatum of dSTR-Shati OE mice. We also investigated the contribution of projected 5-HTergic neuron from dorsal raphe nucleus to dorsal striatum to stress sensitivity using DREADD system. [Results] We found that 5-HT levels in the dorsal striatum decreased in stress susceptible, but not resilient, mice, and these levels correlates with social interaction behaviors. dSTR-Shati OE mice showed the vulnerability to even microdefeat stress and the reduction of striatal 5-HT levels. The vulnerability to stress in Shati OE mice recovered by microinjection of SSRI into the dorsal stratum or activation of dorsal raphe nucleus-dorsal striatum 5-HTergic neuron by DREADD system. [Conclusions] Striatal Shati/Nat8l controls stress sensitivity via regulation of dorsal striatum-dorsal raphe nucleus circuit resulting in control of 5-HT levels in the dorsal striatum. Our study suggests the novel mechanism underlying stress sensitivity in the depression pathogenesis, and striatal Shati/Nat8l could be a novel target for medical tools for depression. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-276
C57BL/6J系マウスにおけるデキストラン硫酸ナトリウム誘発性大腸炎は慢性予測不可軽度ストレスに対する感受性を増大させる
Dextran sulfate sodium-induced colitis in C57BL/6J mice increases their susceptibility to chronic unpredictable mild stress that induces depressive-like behavior
*長澤 一樹(1)、河本 実季(1)、浅田 あゆみ(1)、大嶋 廉之(1)、宮永 佳代(1)、森本 博俊(2)、安川 岳志(2)、森戸 克弥(1)、魚住 嘉伸(2)、髙山 健太郎(1)
1. 京都薬科大学、2. 赤穂化成
*Kazuki Nagasawa(1), Miki Komoto(1), Ayumi Asada(1), Yasuyuki Ohshima(1), Kayo Miyanaga(1), Hirotoshi Morimoto(2), Takeshi Yasukawa(2), Katsuya Morito(1), Yoshinobu Uozumi(2), Kentaro Takayama(1)
1. Kyoto Pharmaceutical University, 2. Ako Kasei, Co., Ltd
Keyword: depressive-like behavior, chronic unpredictable mild stress, colitis
Objective: In patients with colitis, the high comorbidity of depressive disorders is well-known, but the detailed mechanisms remain unresolved. In this study, we examined whether colitis induced by dextran sulfate sodium (DSS) increased the susceptibility to chronic unpredictable mild stress (CUMS) in C57BL/6J mice with resilience to CUMS.
Materials & Methods: To induce experimental colitis and depressive-like behaviors, male 7-weeks old C57BL/6J mice were administered ad libitum 1% DSS solution for 11 days, and subjected to various mild stressors in a chronic, inevitable and unpredictable way according to a random schedule for 21 days, respectively.
Results & Discussion: In naïve mice exposed to CUMS, their immobility times in a forced swim (FS) test were almost equal to those in control mice. The DSS administration to naïve mice induced colitis without depressive-like behavior, and at 18 days after termination of the DSS administration, the colitis had recovered to control levels, while altered diversity and composition of bacterial genera such as Bacteroides spp., Alistipes spp., etc., were found in the gut microbiota. Exposure of mice with DSS-induced colitis to CUMS (DSS+CUMS) significantly increased the immobility times in the FS test. In the gut microbiota of DSS+CUMS mice, the alteration profile of the relative abundance of bacterial genera differed from in the DSS ones. These findings indicate that mice with colitis exhibit increased susceptibility to psychological stress, resulting in induction of depressive-like behavior, and this might be due, at least in part, to altered characteristics of the gut microbiota. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-277
身体的ストレスを軽減させたプロトコルによる社会的敗北ストレスモデルマウスの確立とその行動解析
Development and behavioral characterization of micro social defeat-models using male BALB/c mice.
*西澤 秀朗(1)、谷島 優平(1)、豊田 淳(1,2)
1. 茨城大学農学部、2. 東京農工大学大学院連合農学研究科
*Hideaki Nishizawa(1), Yuhei Yajima(1), Atsushi Toyoda(1,2)
1. College of Agriculture, Ibaraki University, 2. United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology
Keyword: BALB/c , Depression, Nest building, Social defeat
[Introduction]
Socially defeated mice are widely used for preclinical studies of depression. The social defeat paradigm involves both physical and psychological stresses that occurred between aggressors and subordinates, and unfortunately, excessive physical attacks by aggressors cause severe injuries to subordinates. These physical damages make inconsistency in the preclinical models of depression; therefore, reduction of physical contacts is needed for an appropriate paradigm to produce socially defeated mice. This study aimed to develop two types of socially defeated male BALB/c mice subjected to mild physical stress compared with the standard protocol (Golden et al., Nat Protoc, 2011), and characterized several behaviors in these models.
[Methods] BALB/c mice (male, 7 weeks) were purchased and habituated in our animal facility for 1 week. The social defeat paradigm included 10-days sessions with physical and psychological stresses using male ICR mice as aggressors. BALB/c mice were divided into 3 groups, namely the group subjected to subchronic and micro social defeat stress (sMSDS), the group subjected to subchronic and mild social defeat stress (sCSDS) (Goto et al., 2014), and the group without stress (Control). In sMSDS, the duration of physical contacts between BALB/c and ICR was set at 1 min after the first attack bite by ICR. After physical contact, BALB/c were moved into the neighboring compartment to the ICR and housed for 24 hours. To evaluate physical contacts, the number of bites was counted manually. For evaluation of depressive- and anxiety-like behaviors in mice, various behavior tests were performed, namely nest building, social interaction, elevated-plus maze (EPM), forced-swimming (FS), and tail suspension (TS).
[Results and discussion] Total feed intake and body weight gain were not significantly different between the three groups. Defeat-induced polydipsia was not observed in BALB/c mice subjected to sMSDS and sCSDS, while sCSDS induced polydipsia in C57BL/6J male mice (Goto et al., Behav Brain Res, 2014). sMSDS and sCSDS increased weights of several tissues including the spleen and cecum compared with control. As our previous study showed the deficits of nest building in B6 mice subjected to sCSDS and acute social defeat stress (Otabi et al., Behav Proc, 2016; 2017), BALB/c mice subjected to sMSDS and sCSDS showed deficits of nest building. The social interaction test with ICR in the open field arena revealed that the rates of susceptible mice were 17, 63, and 67% of control and sMSDS, sCSDS mice respectively. Other behavioral analyses such as EPM, FS, and TS revealed that there is no significant difference between the three groups. In conclusion, sMSDS could be a useful stress paradigm to produce a mild depression model with less physical damage. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-278
前頭前皮質のIDO1は慢性社会的敗北ストレスに誘発されるうつ様行動を制御する
IDO1 in the prefrontal cortex regulates depression-like behavior following chronic social defeat stress
*國澤 和生(1)、小菅 愛加(1)、飯田 翼(1)、河合 智貴(1)、田辺 萌夏(1)、齋藤 邦明(2,3,4)、鍋島 俊隆(3,4)、毛利 彰宏(1,4)
1. 藤田医科大学大学院保健学研究科・レギュラトリーサイエンス分野、2. 藤田医科大学大学院保健学研究科・先進診断システム開発分野、3. 藤田医科大学大学院保健学研究科・先進診断システム探索部門、4. NPO法人医薬品適正使用推進機構
*Kazuo Kunisawa(1), Aika Kosuge(1), Tsubasa Iida(1), Tomoki Kawai(1), Moeka Tanabe(1), Kuniaki Saito(2,3,4), Toshitaka Nabeshima(3,4), Akihiro Mouri(1,4)
1. Dept. Regulatory Sci., Grad. Sch. Health Sci., Fujita Health Univ., Aichi, Japan, 2. Dept. Adv Diagnostic Syst Dev., Grad. Sch. Health Sci., Fujita Health Univ., Aichi, Japan, 3. Adv Diagnostic Syst Res Lab., Grad. Sch. Health Sci., Fujita Health Univ., Aichi, Japan, 4. NPO. J-DO., Aichi, Japan
Keyword: Depression, Stress, IDO1, Tryptophan pathway
Indoleamine 2,3-dioxygenase 1 (IDO1) metabolizes the essential amino acid tryptophan into kynurenine (KYN), which is involved in neural activity via the KYN pathway. It has been proposed that IDO1 implicates in the pathogenesis of major depressive disorder (MDD). However, the underlying mechanisms remain poorly understood. In the present study, we investigated an involvement of KYN synthesis in chronic social defeat stress (CSDS), a mouse model of depression. In the CSDS, C57BL/J mice were exposed to aggressor ICR mouse for 10 consecutive days. CSDS persistently reduced the duration of time spent at the interaction zone in the social interaction test 4 weeks after the last stress exposure. We found reduced expression of IDO1 and KYN concentration in the prefrontal cortex of mice 4 weeks, but not 1 day after the last stress exposure. Double immunostaining showed the IDO1 immunoreactivity was mainly observed in neurons of the prefrontal cortex of control mice, and which was reduced in the mice exposed to CSDS. To explore whether IDO1-mediated KYN reduction in mice exposed to the stress is involved in persistent social impairments, KYN (50 mg/kg; i.p.) was administrated to the stressed mice from 1 day after the last stress exposure for 4 weeks. KYN attenuated the impairment of social behaviors. Furthermore, injection of adeno-associated virus (AAV) expressing shRNA against IDO1 into the prefrontal cortex increased susceptibility to persistent social impairments induced by a subthreshold CSDS for 3 days. These results suggest that IDO1 may be a critical regulator of stress vulnerability and KYN could be a potential target for the development of antidepressants. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-279
マウス海馬における過成熟現象
Transcriptomic evidence of hyper-maturation in the mouse hippocampus induced by genetic and other factors
*萩原 英雄(1)、小清水 久嗣(1)、服部 聡子(1)、田中 美歩(2)、池田 和隆(2)、宮川 剛(1)
1. 藤田医科大学 総合医科学研究所 システム医科学研究部門、2. 東京都医学総合研究所 精神行動医学研究分野 依存性物質プロジェクト
*Hideo Hagihara(1), Hisatsugu Koshimizu(1), Satoko Hattori(1), Miho Tanaka(2), Kazutaka Ikeda(2), Tsuyoshi Miyakawa(1)
1. Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Aichi, Japan, 2. Addictive Substance Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan
Keyword: Transcriptome, Hyper-maturation, Hippocampus
Adequate maturation of neurons and the relevant synaptic organization in the hippocampus are indispensable for normal cognitive function and emotional behavior. Accumulating evidence has suggested that maturation abnormalities of hippocampal principal neurons and other brain regions/cell types have been implicated in multiple neuropsychiatric disorders and the model animals. To date, most studies on maturation abnormality phenomena have involved immaturity, in which maturation status is arrested at or reversed to pseudo-immature state in adulthood due to various genetic and acquired factors. Knowledge on opposite phenomena, or hyper-maturity in the brain, is limited. In this study, by screening the database containing more than 128,000 RNA expression datasets, we provide transcriptomic evidence of hyper-maturation of the hippocampus in 14 different mouse strains and mice treated with experimental manipulations. Hyper-maturation in those strains/conditions of mice (mutant/experimental mice vs. wild-type/control mice) was defined to show a high negative correlation with typically developing infant mice (infants vs. adults) in the comparison of genome-wide gene expression patterns. The strains/conditions of mice with hyper-maturation included such as serotonin transporter knockout mice, glucocorticoid receptor over-expression transgenic mice, mouse models of depressive/anxiety disorders, Df(16)A/+ mice, 22q11.2 deletion mouse model of schizophrenia, mice chronically treated with the PDE9A inhibitor, and hypoxia-induced mice. Meta-analysis of the 14 datasets highlighted that feature genes involving hyper-maturation were predominantly enriched for synapse-associated genes. This study suggests that hyper-maturation of the hippocampus can be induced by various factors, especially accompanied by the altered transcriptional organization of synapse-related genes. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-280
ケタミンの抗うつ作用には内側前頭前皮質の下流の異なる投射標的が関与する
Distinct downstream targets of the medial prefrontal cortex underlie discrete antidepressant responses to ketamine
*篠原 亮太(1,2)、Brendan D Hare(2)、Rong-Jian Liu(2)、Jin Hua Li(2)、Xiao-Yuan Li(2)、Catharine H Duman(2)、Ralph J DiLeone(2)、Ronald S Duman(2)
1. 神戸大学大学院医学研究科薬理学分野、2. Yale大学医学部分子精神医学部門
*Ryota Shinohara(1,2), Brendan D Hare(2), Rong-Jian Liu(2), Jin Hua Li(2), Xiao-Yuan Li(2), Catharine H Duman(2), Ralph J DiLeone(2), Ronald S Duman(2)
1. Div Pharmacol, Grad Sch Med, Kobe Univ, Kobe, Japan, 2. Dept Psychiatry, Yale Sch Med, New Haven, USA
Keyword: KETAMINE, MAJOR DEPRESSIVE DISORDER, ANTIDEPRESSANTS, NEURAL CIRCUITS
Ketamine, an N-methyl-D-aspartate (NMDA) receptor antagonist, is the prototype for a potential new generation of glutamate-based antidepressants that rapidly relieve symptoms of depression within hours of treatment. Studies in rodents have demonstrated that neuroplasticity in the medial prefrontal cortex (mPFC) is critical for the antidepressant actions of ketamine. However, effector circuits downstream of the mPFC underlying the rapid antidepressant responses remain unknown. To address this issue, we used optogenetic and chemogenetic circuit mapping in rodent models for studying depression, demonstrating the role of the basolateral amygdala (BLA) and bed nucleus of stria terminalis (BNST) as downstream targets of the mPFC mediating distinct behavioral effects of ketamine. Activation of BLA principal neurons by mPFC inputs is critical for a subset of ketamine’s antidepressant-like effects. An antidepressant dose of ketamine selectively activates BLA neurons projecting to the ventral hippocampus and increases dendritic spines and excitatory transmission in these neurons. Activation of BLA projections to the ventral hippocampus is sufficient to produce persistent antidepressant-like effects on passive coping behavior but not on anxiety-like and reward-seeking behaviors. In contrast, mPFC projections to the BNST are necessary and sufficient to produce persistent antidepressant-like effects on anxiety-like and reward-seeking behaviors but not on passive coping behavior. Therefore, our data support a model where distinct downstream circuits of the mPFC contribute to producing separate antidepressant-like behavioral responses. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-281
社会的敗北ストレスモデルマウスの糞便、血漿からストレスバイオマーカー探索の試み
Identification of stress biomarkers in feces and blood plasma of the social defeat model of mouse
*谷島 優平(1)、中村 勇(2)、川瀬 貴博(3)、塚原 隆充(3)、豊田 淳(1,4)
1. 茨城大学農学部、2. 茨城県立医療大学保健医療学部、3. 株式会社栄養・病理学研究所、4. 東京農工大学大学院連合農学研究科
*Yuhei Yajima(1), Isamu Nakamura(2), Takahiro Kawase(3), Takamitsu Tsukahara(3), Atsushi Toyoda(1,4)
1. Col of Agr Ibaraki Univ, Ibaraki, Japan, 2. Ctr for Med Sci, Ibaraki Pref Univ of Health Sci, Ibaraki, Japan, 3. Kyoto Inst of Nutr & Pathol Inc, Kyoto, Japan, 4. United Grad Sch of Agr Sci, Tokyo Univ of Agri and Tech, Tokyo, Japan
Keyword: Biomarker, Metabolite, Microbiome, Social defeat stress
[Introduction]
Discoveries of stress biomarkers, especially early and noninvasive biomarkers for depressive disorders are essential for establishing clinical strategies to prevent depression. Unfortunately, it would be difficult to identify these early biomarkers in the clinical samples, thus intensive research using animal models of depression should be conducted. Previously, we discovered psychosocial stress influences peripheral metabolome and gut microbiota in a mild model of depression, namely a subchronic and mild social defeat stress (sCSDS) model of mice (Goto et al., J Proteome Res., 2015; Aoki et al., J Proteome Res., 2016). In this study, we aimed to identify critical biomarkers for stress susceptibility or resilience from noninvasive samples such as feces and blood plasma in sCSDS mice using microbiome and metabolome analyses.
[Methods]
sCSDS paradigm was described in our previous paper (Goto et al., Behav Brain Res., 2014). C57BL/6J mice (B6, male, 8 weeks) were divided into 2 groups subjected to no stress (control) or sCSDS. After 10-days of the sCSDS period, we conducted social interaction (SI) test to confirm stress vulnerability in B6 and succeeded to obtain susceptible and resilient mice based on their SI scores. Data from three groups (control, susceptible, and resilient) were characterized. Feces from pre-sCSDS and post-sCSDS were characterized for microbial metagenome and metabolome using next-generation sequencing and GC-MS, respectively. Moreover, blood plasma from post-sCSDS was analyzed for metabolome using GC-MS/MS. All mice were fed AIN-93G diets ad libitum. Data analyses were conducted by R, QIME, and MetaboAnalyst.
[Results and discussion] Metagenome analysis revealed that sCSDS increases α diversity and affect the composition of gut microbiota, and some specific gut microbes were enriched in each group. Metabolome analysis for feces revealed that sCSDS affects levels of several metabolites linked to purine and glucose metabolism. Collectively, control, susceptible, and resilient mice had unique gastrointestinal ecosystems. Furthermore, sCSDS affected plasma metabolome and changed levels of 49 metabolites, and most metabolites were downregulated by sCSDS. On the other hand, the level of plasma 4-hydroxyproline was increased by sCSDS, which is consistent with our previous study (Goto et al., J Proteome Res., 2017). Precise information about potential biomarkers for stress vulnerability will be discussed in the meeting. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-282
うつ病と感情経験時における内受容感覚知覚の関係―心拍誘発電位を用いた検討
Interoceptive awareness in Depression during emotional experiences: An examination using heartbeat evoked potentials
*荻島 大凱(1)、嶋田 洋徳(2)
1. 高知工科大学フューチャー・デザイン研究所、2. 早稲田大学人間科学学術院
*HIROYOSHI OGISHIMA(1), Hironori Shimada(2)
1. Research Institute of Future Design, Kochi University of Technology, Kochi, Japan, 2. 2)Faculty of Human Sciences, Waseda University, Saitama, Japan
Keyword: DEPRESSION, INTEROCEPTION, HEARTBEAT EVOKED POTENTIAL
Background. Depression is a mental disorder which associated with difficulties in emotional regulation. Recently, these features have been understood to be associated with interoception, which refers to the internal senses of the body. According to this view, depressed patients are prone to lower interoceptive awareness. This leads to insensitivity to changes in their biological state, including their emotional responses. Consequently, difficulties in easing unpleasant moods and in experiencing pleasant moods are exhibited. Previous studies have, however, not examined interoceptive awareness in depression when a specific emotional state is induced. Therefore, this hypothesis has not been sufficiently examined. In this study, we measured heartbeat evoked potentials and event-related potentials of heartbeats during mood inductions to investigate interoceptive awareness in depression, taking emotional valence into account.
Methods. A total of 62 undergraduate and graduate students participated in the study. After they were screened for depressive symptoms by means of the CES-D, they were experimentally induced to experience pleasant, unpleasant, and aroused states. Their heart-rate evoked potentials were measured during these inductions. For the induction of pleasant and unpleasant feelings, an induction task was performed in which image and sound stimuli were presented in pairs. For the induction of an aroused feeling, a modified version of the Montreal Imaging Stress Task, a time-limited mental calculation task, was used. Discriminant analysis was performed on each individual's increase/decrease in values of heart-rate evoked potentials during pleasant, unpleasant, and aroused mood inductions, and plotted on a three-dimensional Euclidean space.
Results. The results revealed that the heart-rate evoked potentials of the individuals who were depressed (22 females and 9 males, mean age = 22.44 ± 2.90 years) were attenuated during the induction of the pleasant mood, compared to those who were not depressed (20 females and 11 males, mean age = 22.41 ± 3.28 years) (F (1, 61) = 0.33, P = .039). Discriminant analysis indicated that decreased heart-rate evoked potentials during a pleasant mood induction could predict depression in participants.
Discussion. These results indicate that participants with severe depressive symptoms have difficulty perceiving their internal bodily states, especially during pleasant mood experiences. In other words, depression may be a state in which it is difficult to integrate bodily experiences into the processes of emotional experience, only when the specific emotion of pleasure is experienced. Therefore, a depressive mood may be understood to be associated not only with the presence of cognitive distortions, but also with a feature of emotional processing, namely the inability to integrate bodily experiences with emotional experiences. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-283
ニュース聴取時の脳波に基づく抑うつ気分の推定
Detection of depressive mood states based on EEG data during news listening
*渡部 宏樹(1)、伏田 幸平(1)、齊藤 隼平(1)、成瀬 康(1)、井原 綾(1)
1. 国立研究開発法人情報通信研究機構
*Hiroki Watanabe(1), Kohei Fuseda(1), Jyunpei Saito(1), Yasushi Naruse(1), Aya S Ihara(1)
1. National Institute of Information and Communications Technology
Keyword: Depressive mood, EEG, speech, language
In a recent information-overloaded society, there are more opportunities to receive negative information in our daily lives, increasing the risk of mental health problems such as depression. Although it has been found that depressed mood can be detected using resting-state electroencephalogram (EEG), there is little research on whether it can be detected from daily activities. In this study, we focused on news listening as one of the daily activities and aimed to clarify whether EEG features during listening to auditory news with negative, neutral, and positive contents are useful for detecting depressed mood in a non-clinical population. Since depressed patients show attentional bias toward negative stimuli, attention to negative news and its semantic processing would be expected to be modulated by the mood state. Thus, we evaluated the classification performance of individuals with depressed mood using the mean amplitude and peak latency of N1, P2 (auditory processing; modulated by attention), and N400 (semantic processing) time-locked to word onset in each news.
The healthy 135 native Japanese speakers (69 females, age range = 20–49 years old) listened to 30 radio news clips (about 1 minute, 15 clips were used for analysis) with negative, neutral, and positive content. Their depressed mood was assessed with the Beck Depression Inventory-II (depressed mood group = 32 participants). A temporal response function was used to estimate each participant’s event-related potentials time-locked to the onset of each word in the news. The mean amplitudes and peak latencies of N1, P2, and N400 were calculated for each electrode (Fpz, Cz, and Pz) and news content. Based on the statistical analysis, we prepared 12 features (peak latencies of N1 and P2 at Fpz and mean amplitudes of N1 and N400 at Pz for each news content type). Six features were selected from them using the recursive feature elimination method. A linear support vector machine was trained and evaluated by the leave-one-out cross-validation.
As a result, the area under the receiver operating characteristic curve achieved 0.73. The correct classification rates for the depressed and non-depressed mood group were 0.656 (21/32 participants) and 0.660 (68/103 participants), respectively, suggesting that EEG features during news listening are effective in detecting depressed mood and that measurement can lead to early detection of depressive tendencies in daily life. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-284
深層学習に基づいた動物姿勢解析ソフトウェアのtail-suspension testへの応用
Application of a deep learning-based animal-pose analysis software for tail-suspension test
*古瀬 民生(1)、串田 知子(1)、西村 静佳(1)、山田 郁子(1)、田村 勝(1)
1. 理化学研究所 バイオリソース研究センター
*Tamio FURUSE Furuse(1), Tomoko Kushida(1), Shizuka Nishimura(1), Ikuko Yamada(1), Masaru Tamura(1)
1. RIKEN BioResource Research Center
Keyword: Tail suspension test, Deep learning, Mouse phenotyping
In the last 10-20 years, quantification of animal behavior in laboratory has been automated by computer software. The software are available from commercial suppliers, and they have advantage in terms of usability such as graphical user interface (GUI). In addition, well-used commercial software have advantages in the terms of the objectivity of measured parameters by comparing data collected in different laboratories. On the other hand, potential of scalability of test parameters in each behavioral test is limited in the commercial software. Recently, artificial intelligence-based software for analyzing animal behavior are freely available in research community. A recent study (Sturman et al. 2020) revealed that an accuracy of behavioral detection by a deep learning-based software, DeepLabCut (Mathis et al. 2018) reaches human accuracy in elevated plus maze test. Coordination of animals’ body parts in video files are automatically detected by the DeepLabCut. Behavioral parameters in the elevated plus maze were calculated by using the coordination of the body parts in the previous study (Sturman et al. 2022). In order to carry out high throughput mouse phenotyping, the commercial behavioral-quantification software is used also in the Japan Mouse Clinic (JMC) at the RIKEN BioResource Research Center. The JMC provides research communities in Japan with a comprehensive-behavioral phenotyping system. The comprehensive-behavioral phenotyping system is named Pipeline 2. On the other hand, we perform behavioral tests not included in the Pipeline 2 in order to carry out deep phenotyping of mutant mice. In the Pipeline 2, the DeepLabCut is used in several tests such as tail-suspension test. Prior to phenotyping of mutants, we carried out the tail suspension test by using 5 mouse strains, C57BL/6N, C57BL/6N, DBA/2J, BALB/cA, and C3H/HeJ. We will report difference of a depression-like behavioral parameter, immobility time, measured by using the DeepLabCut in mouse strains above. References Sturman et al. Deep learning-based behavioral analysis reaches human accuracy and is capable of outperforming commercial solutions. Neuropsychopharmacology. 2020 45(11):1942-1952. Mathis et al. DeepLabCut: markerless pose estimation of user-defined body parts with deep learning. Nat Neurosci. 2018 21(9):1281-1289. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-285
脳-代謝連関：糖尿病モデルマウスにおけるうつ病様行動増悪の分子メカニズム
Brain-Metabolic axis: Molecular mechanism of exacerbated depression-like behaviors in a diabetes mellitus model
*天間 雄祐(1)、三宅 隆平(1)、小尾（永田） 紀翔(1)、片山 雄太(2)、萩原 英雄(3)、宮川 剛(3)、中山 敬一(2)、林（高木） 朗子(1)
1. 理研CBS・多階層精神疾患研究チーム、2. 九州大学・ 生体防御医学研究所・分子医科学分野、3. 藤田医科大学・総合医科学研究所・システム医科学研究部門
*Yusuke Temma(1), Ryuhei Miyake(1), Kisho Obi-Nagata(1), Yuta Katayama(2), Hideo Hagihara(3), Tsuyoshi Miyakawa(3), Keiichi I Nakayama(2), Akiko Hayashi-Takagi(1)
1. Lab for Multi-scale Biological Psychiatry, RIKEN CBS, Saitama, Japan, 2. Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan, 3. Division of Systems Medical Science, Institute for Comprehensive Medical Science, Fujita Health University, Aichi, Japan
Keyword: Anhednia, Diabetes mellitus, Chronic restraint stress
Lifestyle disturbances often accompany psychological stress, and the comorbidity of depression and lifestyle-related diseases, such as diabetes mellitus (DM), is currently a serious problem. Epidemiological evidence has suggested that the comorbidity of DM and depression has a greater health impact than the simple sum of the individual conditions and that these diseases mutually exacerbate their pathophysiology and prognosis. However, the underlying molecular mechanism of the interaction of each disorder remains unclear. To address this unmet medical question, we generated four animal models: no treatment (control group), chronic restraint stress as a depression model (CRS group), streptozotocin administration as a diabetes model (DM group), and a combination of CRS and streptozotocin administration as a comorbid of two conditions (comorbid group). A series of behavioral analyses revealed that the comorbid group exhibited a significantly severer anhedonia-like behavior, as shown by the female encounter test. To elucidate the molecular mechanism of this interaction, we performed gene set enrichment analysis (GSEA) of the brain from four groups and found that pathways involved in pH homeostasis and inflammation were significantly altered in the comorbid group with a significant CRS x DM interaction effect. Interestingly, we found the brain pH significantly correlated with depression-like behavior, suggesting that lower brain pH and inflammation can contribute to the exacerbating depression-like behavior in the comorbid model. Taken together, the pH homeostasis and inflammation can be involved in the susceptibility of DM to depression, and ameliorating abnormal pH and inflammation of the diabetic brain may be novel treatment strategies for depressive-related conditions. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-286
繰返し社会的敗北ストレス負荷マウスの抑うつ様行動と帯状皮質Arc陽性細胞の活動に対するケタミンの効果
Effects of ketamine on chronic social defeat stress-induced depressive-like behaviors and Arc-positive cell activity of the cingulate cortex in mice
*二階堂 義和(1,2)、緑川 陽子(2)、櫛方 哲也(2)、廣田 和美(2)
1. 弘前大学大学院医学研究フレイル予防学研究講座、2. 弘前大学大学院医学研究科麻酔科学講座
*Yoshikazu Nikaido(1,2), Yoko Midorikawa(2), Tetsuya Kushikata(2), Kazuyoshi Hirota(2)
1. Dept Frail Res Prevent, Hirosaki Univ, Hirosaki, Japan, 2. Dept Anesth, Hirosaki Univ, Hirosaki, Japan
Keyword: Ketamine , Depression, Social defeat stress, Activity-regulated cytoskeleton-associated protein (Arc)
Ketamine, a non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonist, is known as an intravenous anesthetic. Major depression is associated with widespread neural and behavioral dysfunctions. Recent clinical research showed that sub-anesthetic dose ketamine has a rapid antidepressant effect in patients with major depression. The antidepressant effects of ketamine might be partly attributed to the neural activity of the prefrontal cortex; however, it remains unknown how the antidepressant ketamine influences cortical neural activity. In this study, we examined the effect of low-dose ketamine (10 mg/kg i.p.) on immediate-early gene product activity-regulated cytoskeleton-associated protein (Arc)-positive cells in the cingulate cortex of C57BL/6J mice following chronic social defeat stress (CSDS). After CSDS, depression-susceptible mice showed social avoidance, reduction of sucrose preference, and decreases in the number of Arc-positive cells in the cingulate cortex (Arc-positive cells count/mm2, mean ± SD: control, 186 ± 40.0; resilient, 206 ± 70.2; susceptible, 26.1 ± 20.3; P < 0.05, vs. control and resilient). Low-dose ketamine administration improved these behavioral and neuronal alterations of depression-susceptible mice (susceptible+ketamine, 196 ± 73.1; P < 0.05, vs. susceptible). To confirm the effects of ketamine on the Arc-positive cingulate neurons, we performed in vivo Ca2+ imaging of GCaMP6s with Arc-CreERT2 mice and adeno-associated virus containing a flex-GCaMP6s cassette. Low-dose ketamine increased Ca2+ responses in Arc-positive neurons in the cingulate cortex (Hz: control, 0.008 ± 0.01; ketamine, 0.04 ± 0.02; P < 0.05). These results suggest that Arc-positive neuronal activity in the cingulate cortex might play an essential role in the antidepressant effect of ketamine. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-287
超音波暴露は嗅球摘出ラットのうつ様行動を改善する
Ultrasound exposure improves depressive-like behavior in olfactory bulbectomized rats
*山内 つぐみ(1,2)、吉岡 寿倫(1)、山田 大輔(1)、濱野 匠(1)、市川 寛子(2)、池田 麻維花(6)、亀井 雅人(6)、大槻 貴也(6)、佐藤 康夫(6)、新居 恭子(6)、鈴木 将史(6)、入山 聖史(4)、吉澤 一巳(5)、西野 彰一(6)、宮崎 智(3)、斎藤 顕宜(1)
1. 東京理科大学 薬学部 薬学科 薬理学研究室、2. 東京理科大学 教養教育研究院 野田キャンパス教養部 心理学研究室、3. 東京理科大学 薬学部 薬学科 生命情報科学研究室、4. 東京理科大学 理工学部 情報科学科 量子情報力学研究室、5. 東京理科大学 薬学部 薬学科 疾患薬理学研究室、6. 株式会社フジミック
*Tsugumi Yamauchi(1,2), Toshinori Yoshioka(1), Daisuke Yamada(1), Takumi Hamano(1), Hiroko Ichikawa(2), Maika Ikeda(6), Masato Kamei(6), Takaya Otsuki(6), Yasuo Sato(6), Kyoko Nii(6), Masashi Suzuki(6), Satoshi Iriyama(4), Kazumi Yoshizawa(5), Syoichi Nishino(6), Satoru Miyazaki(3), Akiyoshi Saitoh(1)
1. Lab Pharmacol, Fac Pharm Sci, Tokyo Univ of Science, 2. Lab Psychol., Inst of Arts and Sci, Tokyo Univ of Science, 3. Lab Bioinfo, Fac Pharm Sci, Tokyo Univ of Science, 4. Lab Quant info dynamics., Fac Sci and Tech, Tokyo Univ of Science, 5. Lab Disease Pharmcol., Fac Pharm Sci, Tokyo Univ of Science, 6. FUJIMIC, inc. Tokyo
Keyword: Ultrasonic vocalization, Depression, OBx rat, Animal model
[Introduction] In recent years, the effects of ultrasound on emotions have been attracting attention. Some experiments have shown that exposure to ultrasound on the human body surface activates brain regions of the reward system, known as “hypersonic effect”, which is expected to be applied to the Cognitive behavioral therapy depression. However, the details of the mechanism are still unclear. Therefore, in the present study, we investigated the effects of ultrasound exposure to elucidate its mechanism using olfactory bulbectomized (OB) rats, the animal models of depression. [Methods] Male Wistar/ST rats (9-10 weeks) were used. OB rat model was created by removing the olfactory bulb from the rats. Depression-related behavior in OB rats were evaluated according to the hyperemotionality (HE) by scoring. We collected the ultrasonic vocalizations (USV) associated with pleasant emotions (50 kHz-USV) by handling of the male rats. Artificial sounds were generated using Avisoft-SASLob Pro (Avisoft). Rats were exposed to 50 kHz USV or artificial 50 kHz/100 kHz ultrasounds for 24 h in a soundproof box. The HE scores in rats were evaluated before and after exposure to ultrasound. The control group was kept in a soundproof box for 24 hours without being exposed to ultrasound. [Results] The HE scores in OB rats were significantly increased, as compared to those in sham rats. Interestingly, the exposure to 50 kHz-USV significantly reduced HE scores in OB rats. Furthermore, artificial 50 kHz ultrasounds significantly decreased the HE scores in OB rats. On the other hand, both artificial 100 kHz ultrasounds and white noise (35-100 kHz) produce no effects on HE scores in OB rats. Neither 50 kHz USV nor artificial sounds exposure produced no effects on the HE scores in sham rats. [Discussion] These results indicated that exposure of 50 kHz USV and artificial 50 kHz ultrasound produced the antidepressant-like effects in OB rats, suggesting that 50 kHz frequency-range sounds could play an important role in hypersonic effects. Moreover, we propose that the OB rats might be a useful animal model to elucidating hypersonic effects. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-288
大うつ病における皮質下体積と磁化率の解析
Subcortical morphometry and magnetic susceptibility analysis in major depression disorder
*渋川 周平(1,3,4)、菅 博人(2)、舞草 伯秀(1)、小池 進介(1)
1. 東京大学総合文化研究科 進化認知科学研究センター、2. 名古屋大学、3. 順天堂大学、4. 東京医科大学
*Shuhei Shibukawa(1,3,4), Kan Hirohito(2), Norihide Maikusa(1), Koike Shinsuke(1)
1. The University of Tokyo, 2. Nagoya University, 3. Juntendo University, 4. Tokyo Medical University
Keyword: magnetic susceptibility mapping (QSM), major depression disorder (MDD), subcortical volume, susceptibility
Objective
Abnormalities in brain structures including cortical thickness and subcortical volumes are observed in patients with major depression disorder (MDD). Quantitative magnetic susceptibility mapping (QSM) allows detecting subtle changes in magnetic susceptibility. Since the susceptibility value is affected by iron and myelin concentration in the brain, QSM analysis may be considered as a sensitive imaging for understanding subcortical structures. This study aimed to evaluate the differences in subcortical volume and susceptibility distribution between MDD patients and healthy controls.
Methods
We investigated 45 patients with MDD (mean age = 39.1 years, 21 females) and 20 healthy age- and gender-matched controls (mean age = 48.6 years, 10 females). All participants underwent 3.0 Tesla brain magnetic resonance imaging (MRI) including QSM 3D T1-weighted image. We subdivided the subcortical structures and measured the volumes and the susceptibility in the red nucleus, substantia nigra, caudate, putamen, thalamus, and globus pallidus externa and interna. The statistical significance between the groups was assessed using paired t-tests with false discovery rate correction.
Results and Discussion
Subcortical volume in any brain region did not differ significantly between the groups (P > 0.05). Similar increase of the susceptibility values was found in the red nucleus, substantia nigra, globus pallidus externa, and globus pallidus interna compared with other subcortical structures, but no significant group difference in the susceptibility value was observed (P > 0.05). In MDD groups, there were no structural changes that altered subcortical volume or susceptibility compared with healthy controls. We will validate other psychiatric disorders in the future study.
Conclusions
This study found no significant differences in subcortical volume and susceptibility distribution were detected between MDD patients and healthy controls. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-289
逆トランスレーショナルアプローチによる双極性障害の新規動物モデル
A novel animal model of bipolar disorder using a reverse translational approach
*太田 航(1)、實木 亨(2)、山ノ上 友美(1)、宮﨑 智之(1)、中島 和希(1)、波多野 真依(1)、佐野 亜加根(1)、谷 英明(3)、⻑井 信弘(3)、小泉 輝樹(3)、中島 振⼀郎(3)、三村 將(3)、内田 裕之(3)、高橋 琢哉(1)
1. 横浜市⽴⼤学医学部 ⽣理学教室、2. 三重大学医学部 生化学教室、3. 慶應義塾⼤学医学部 精神・神経科学教室
*Wataru Ota(1), Susumu Jitsuki(2), Tomomi Yamanoue(1), Tomoyuki Miyazaki(1), Waki Nakajima(1), Mai Hatano(1), Akane Sano(1), Hideaki Tani(3), Nobuhiro Nagai(3), Teruki Koizumi(3), Shinichiro Nakajima(3), Masaru Mimura(3), Hiroyuki Uchida(3), Takuya Takahashi(1)
1. Dept. Physiol., Yokohama City Univ. Sch. of Med., Kanagawa, Japan, 2. Dept. Biochem., Mie Univ. Fac. of Med., Mie, Japan, 3. Dept. Neuropsychiat., Keio Univ. Sch. of Med., Tokyo, Japan
Keyword: AMPA receptors, Cerebellum, Bipolar disorder, mania-like behavior
Bipolar disorder is a life-threatening mood disorder defined by the presence of one or more manic episodes (e.g., abnormally elevated mood) with/without depressive episodes. However, the biological basis of bipolar disorder is poorly understood. Moreover, the effective biological marker to diagnose bipolar disorder has not been established. Recently, we have developed a novel positron emission tomography (PET) tracer for AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptors (Miyazaki et al., Nat. Med., 2020). This tracer exhibited the decrease of AMPA receptors in the cerebellum of bipolar patients was significantly correlated with the symptomatology score of manic state (Young mania rating scale; YMRS). Based on this clinical data, we generated a novel mouse model in which AMPA receptor expression is reduced specifically in the cerebellum. In the last annual meeting of the Japan Neuroscience Society, we have reported that this mice model showed significantly lower immobility time in forced swim test and tail suspension test (i.e., bipolar mania-like behavior). In this presentation, we will show some more data that reinforce the validity of this novel animal model. The mice showed increased sucrose preference, which likely reflect the “hedonic state” usually observed in the manic state of bipolar disorder. It is well known that many circadian functions are altered in people suffering from bipolar disorder. We also examined activity rhythm of the mice model and observed disrupted pattern of locomotor activities. These results suggest a high validity of this novel animal model and are expected to lead to further understanding of the neurobiology of bipolar disorder. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-290
異なるストレスが脳内転写因子活性に与える影響の解析
Comparison of the activity profiles of transcription factors in the brains of mice experienced different stressors
*山本 創(1)、原 龍一郎(1)、安部 健太郎(1)
1. 東北大学大学院 生命科学研究科
*Hajime Yamamoto(1), Ryoichiro Hara(1), Kentaro Abe(1)
1. Tohoku University, Graduate School of Life Science
Keyword: social defeat stress, exercise, transcription factor
Repeated stressful events alter the gene expression pattern of the brain, resulting in adverse symptoms such as those observed in depression. Interestingly, however, repeated stress is also known to improve brain functions, depending on the context and the type of stressor. The mechanisms of how different stressors lead to the distinct phenotype and what determines the good or bad effects of stress on brain function remain unclear. To clear the effect of different stressors on the brain, we obtained the activity profiles of transcription factors, which directly influence the gene expression pattern in the brain. We compared the profile of transcription-factor activity in the brains of mice (Mus musculus) who experienced two types of repeated stress and explored the pivotal transcription factor whose activity relates to the behavioral outcomes of the stress. In this study, we analyzed the chronically defeated or forced-exercised model; mice were given a 10-day repeated social defeat stress, or a 10-day forced rotarod exercise. We collected the brain samples of both mice to analyze the activities of transcription factors by using viral-vector-based transcription-factor activity. The activities of transcription factors are measured by quantitative RT-PCR analysis of the reporter genes. We found that many transcription factors showed chronic changes in their activities for both stressors. From the comparison between the defeated and forced-exercised mice, we found that some transcription factors reflect the effect of stress reversely in their activity, indicating that genes regulated by those transcription factors may change their expression according to the stressed experience. We speculate that such transcription factors showing the experience-dependent change in their activity play essential roles in brain function to converse the behavioral outcome after the stressed experience. This study will help obtain a transcriptional landscape of the effect of stress on the brain and understand its mechanism. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-291
Social defeat-induced cognitive impairment : The effect of dietary restriction
*HSIAO JOU TUNG(1), PEI YU WANG(1)
1. Grad Inst of Brain and Mind Sci, Natl Taiwan Univ, Coll of Med, Taipei, Taiwan
Keyword: dietary restriction, gut microbiome, learning and memory, chronic social defeat stress
Depression is a thorny psychiatric disorder caused by chronic stress. Negative emotion and cognitive impairment are major symptoms of depression that strongly affect more than 300 million people worldwide. Current strategies for treating depression, such as selective serotonin reuptake inhibitors and ketamine-based antidepressant can only improve the emotional response but less effective on the other associated cognitive impairment. Developing a promising intervention that can treat different aspects of disease is therefore urgently needed.
Our previous study showed that dietary restriction (DR, defined as 60% of daily food intake of ad libitum, AL) can significantly improve behavioral outcomes and enhance memory performance of mice through the microbiota-gut-brain axis. However, the impact of DR on depression and associated cognitive impairments are largely unknown. Here, we used chronic social defeat stress model (CSDS) to induce depressive-like behavior and memory dysfunction of mice, and found that 1-month of DR can dramatically improve these behavioral deficits. However, the beneficial effects of DR were abolished when antibiotics were added to the drinking water of CSDS mice. Fecal transplantation of DR-derived microbiota (FMT-DR) to AL CSDS mice was sufficient to mimic DR-induced benefits, further suggesting an important of gut microbiota.
In our future work, we will perform metagenomic sequencing and metabolomics analysis to understand the mechanism underlying DR-induced pro-cognitive effects. We believe this study will make a great impact in the future treatment of mental dysfunction associated with depression. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-292
双極性障害家系iPSニューロンの表現型と原因変異の探索
Cellular phenotypes and causal candidate genes in iPSC-derived neurons of a bipolar disorder family
*高松 岳矢(1,2)、馬目 陽子(4)、小金渕 佳江(7,6)、ディミトロフ ディミタ(8)、當山 奏子(1)、李 俊錫(1,7)、柳 久美子(5)、長谷川 実奈美(4)、早川 朋子(9,1)、近藤 毅(2)、高橋 智幸(8)、要 匡(5)、岡野 ジェイムス洋尚(4)、木村 亮介(3)、松下 正之(1)
1. 大学院医学研究科 分子・細胞生理学講座、2. 琉球大学大学院医学研究科 精神病態医学講座、3. 琉球大学大学院医学研究科 人体解剖学講座、4. 東京慈恵会医科大学 再生医学研究部、5. 国立成育医療研究センター ゲノム医療研究部、6. 東京大学大学院理学系研究科 生物科学専攻、7. 琉球大学医学部 先端医学研究センター、8. 沖縄科学技術大学院大学 細胞分子シナプスユニット、9. 自治医科大学臨床薬理学講座
*Gakuya Takamatsu(1,2), Yoko Manome(4), Kae Koganebuchi(7,6), Dimitar Dimitrov(8), Kanako Toyama(1), Jun-Seok Lee(1,7), Kumiko Yanagi(5), Minami Hasegawa(4), Tomoko Hayakawa(9,1), Tsuyoshi Kondo(2), Takahashi Tomoyuki(8), Tadashi Kaname(5), Hirotaka James Okano(4), Ryosuke Kimura(3), Masayuki Matsushita(1)
1. Dept of Mol Cell Physiol, Grad Sch Med, Univ of the Ryukyus, 2. Dept of Neuropsy, Grad Sch Med, Univ of the Ryukyus, Okinawa, Japan, 3. Dept of Hum Biol Anat, Grad Sch Med, Univ of the Ryukyus, Okinawa, Japan, 4. Div of Regen Med, Jikei Univ Sch of Med, Tokyo, Japan, 5. Dept of Gen Med, Natl Ctr for Chd Hlth Dev, Tokyo, Japan, 6. Dept of Biol Sci, Grad Sch of Sci, Univ of Tokyo, Tokyo, Japan, 7. Adv Med Res Ctr, Fac of Med, Univ of the Ryukyus, Okinawa, Japan, 8. Cel and Mol Syn Func Unit, Okinawa Inst of Sci Tech Grad Univ, Okinawa, Japan, 9. Dept of Phamc, Jichi Med Univ, Tochigi, Japan
Keyword: bipolar disorder, patient-derived iPSC, mitochondria, psychiatry
Bipolar disorder (BD) is a major psychiatric disorder characterized by dysregulation of mood and activity, ranging between depressive and manic states. The pathogenesis is little understood. BD is inheritable with estimated heritability for BD of 70–80%; however, identifying genomic variants that strongly contribute to BD is still unprecedented, and its molecular pathophysiology is almost unknown. To elucidate the pathogenesis of BD, we focused on rare familial patients with potential high-risk genetic factors, and we performed comprehensive genetic analysis and cellular phenotype analysis using patient-derived induced pluripotent stem cells (iPSCs). First, we conducted a pedigree survey in Okinawa, and we found a three-generation multiplex family with BD. We performed parametric linkage analysis, and we detected a significant linkage at a chromosome, and the locus has been reported as a suggestive or significant locus in several previous linkage studies. Then, we generated patient-derived iPSCs from affected individuals of the family. Interestingly, iPSC-derived excitatory neurons from the affected members showed a higher calcium transient frequency than neurons from healthy controls in Fluo4-AM calcium imaging. Moreover, we analyzed allelic imbalances of transcripts in the linkage haplotype by integrating RNA sequencing and whole-genome sequencing. We found haplotype-specific decreased expression of a key mitochondrial regulator gene in the affected individuals of the family. It might contribute to mitochondrial dysfunction and the development of the disease in the family. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-293
アデノ随伴ウイルスを用いたCbln1補充療法はCbln1欠損マウスの小脳失調症状を回復させる
Development of a gene therapy using adeno-associated virus to rescue cerebellar ataxia in Cbln1-defficient mice
*山崎 世和(1)、掛川 渉(1)、柚﨑 通介(1)
1. 慶應義塾大学
*Tokiwa Yamasaki(1), Wataru Kakegawa(1), Michisuke Yuzaki(1)
1. Keio University
Keyword: Cbln1, Synapse, Ataxia, Adeno-associated virus
Cbln1 is expressed and secreted from cerebellar granule cells and serves as a critical synapse organizer for parallel fiber (PF)-Purkinje cell (PC) synapses. Cbln1 forms a tripartite protein complex with its pre- and postsynaptic receptors, neurexin and GluD2, respectively. Deficiency of Cbln1 or GluD2 has been reported to result in severe cerebellar ataxia accompanied by massive loss of PF-PC synapses in both mice and humans. Intriguingly, injection of exogenous Cbln1 proteins into the Cbln1-null cerebellum could rescue the ataxic phenotype by rapidly inducing PF-PC synapses even in the adulthood, indicating Cbln1’s potent synaptogenic activity in vivo. However, the effect of the single injection of Cbln1 proteins was transient, lasting only approximately 1 week. In the present study, we aimed to develop adeno-associated virus (AAV)-based gene therapy to achieve a long-term restoration of Cbln1-deficient mice by continuously supplementing Cbln1. We generated AAV vectors to express Cbln1 under synapsin 1 (Syn), L7 and GFAP promoters in molecular-layer interneurons (MLIs), PCs, astrocytes, respectively. Immunoblot analyses revealed that expression levels of Cbln1 in the Cbln1-null cerebellum infected with AAV-Syn-Cbln1, AAV-L7-Cbln1 or AAV-GFAP-Cbln1 were comparable with those in wild-type mice. Immunohistochemical analyses showed that Cbln1 expressed by AAV-GFAP-Cbln1 was highly localized at PF-PC synapses compared with Cbln1 driven by other promoters. Consistent with these findings, the ataxic phenotype of Cbln1-null mice was most effectively rescued by AAV-GFAP-Cbln1. The effect of GFAP-driven Cbln1 expression was long lasting and could be observed at 6 months after injection. These results suggest that glia cells in the cerebellum is one of the promising targets for an AAV-based gene therapy to restore cerebellar functions in Cbln1-defficient patients. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-294
アンチセンス核酸のin vivo中枢神経毒性の予測を可能とするin vitroアッセイ法の創生
in vitro cytotoxicity assay of antisense oligonucleotides to predict CNS-toxicity in vivo.
*勝山 真帆(1)、吉岡 耕太郎(1)、賈 春燕(1)、Su Su Lei Mon(1)、松林 泰毅(1)、田中 規恵(1)、永田 哲也(1)、横田 隆徳(1)
1. 東京医科歯科大学
*Maho Katsuyama(1), Kotaro Yoshioka(1), Chunyan Jia(1), Su Su Lei Mon(1), Taiki Matsubayashi(1), Kie Tanaka(1), Tetsuya Nagata(1), Takanori Yokota(1)
1. Tokyo Medical and Dental University
Keyword: Antisence oligonucleotide, Nueurotoxicity, Cytotoxicity assay, Lactate dehydrogenase
【Objective】Antisense oligonucleotide (ASO) has attracted attention as a new modality for treatment of intractable neurodegenerative diseases. In 2016, Nusinersen was approved by FDA as the first drug therapy for treatment spinal muscular atrophy by intrathecal administration. However, no ASO targeting CNS diseases has been approved after nusinersen. For clinical application of the ASOs, neurotoxicity induced by ASOs is one of most critical issues. However, the details of the toxicity mechanism have not been clarified and the few ways to predict are known. Ｗe here performed in vivo and in vitro analysis of neuronal toxicity with various ASOs to elucidate the toxicity mechanism and correlation in in vivo and in vitro toxicity.
【Methods】After injections of ASOs which have various modifications, lengths, and target genes into mice via intracerebroventricular (ICV) route, acute CNS toxicity was assessed by behavioral assessment using toxicity scoring (ATSS: Acute Toxicity Scoring System) and open field testing of mice. In vitro assay of cytotoxicity after transfection of those ASOs into neuronal and non-neuronal cell lines, we measured lactate dehydrogenase (LDH) activity released from cells and dehydrogenase activity in viable cells. We analyzed correlation between in vivo neurotoxicity and in vivo cytotoxicity in neuronal cell.
【Results】We confirmed behavioral characteristic of the neurotoxicity induced by several ASOs using toxicity-score based on ATSS and open field test. On ATSS, the neurotoxicity showed decreased arousal and disfunction of motor system. On open field test, the ASOs reduced locomotor activity, resulting in decrease in total distance, maximum speed and mobile time. in vitro assays, in vivo toxic ASOs showed an increase in LDH and a decrease in the number of viable cells in human and mouse neuronal cells, did not in non-neural cells. On the other hand, non-toxic ASO in vivo did not cause cytotoxicity in any cell line. In addition, the in vitro study of chemical modification in the toxic ASO which silence target RNA showed that efficacy of silencing has no correlation neurotoxicity increased of LDH, indicating different structure-activity correlation between the antisense activity and cytotoxicity.
【Conclusion】in vitro evaluation of neuron-dependent cytotoxicity is promising analysis to predict CNS toxicity of ASOs in vivo and to elucidate the mechanism of CNS toxicity. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-295
JigSAP（jigsaw-shaped self-assembling peptide）を用いた亜急性期脳梗塞の再生治療展開
JigSAP (jigsaw-shaped self-assembling peptide) for regenerative medicine of the subacute phase of ischemic stroke
*味岡 逸樹(1,2)、村岡 貴博(2,3)、渡辺 豪(2,4)
1. 東京医科歯科大学、2. 神奈川県立産業技術総合研究所、3. 東京農工大学、4. 北里大学
*Itsuki Ajioka(1,2), Takahiro Muraoka(2,3), Go Watanabe(2,4)
1. Tokyo Medical and Dental U (TMDU), Tokyo, Japan, 2. KISTEC, Kanagawa, Japan, 3. Tokyo University of Agriculture and Technology, Tokyo, Japan, 4. Kitasato University, Kanagawa, Japan
Keyword: Regenerative medicine, Stroke, Artificial scaffolds
Cell-free regenerative therapy requires efficient binding and release of desired growth factors. Although chemically-crosslinked materials can regulate slow release, these materials degrade into chemically-unidentified products in vivo, making them problematic for clinical applications. In contrast, self-assembling peptides that form supramolecular hydrogels are widely used as artificial extracellular matrices (ECMs) in clinical applications because of their cell adhesion properties, scaffold formation, and biodegradability into chemically-identified non-toxic peptides. However, peptide materials that both bind and release growth factors for injured tissue regeneration are critically lacking.
Herein, we combined computational physics, supramolecular chemistry, and molecular biology to create a novel supramolecular peptide material as an artificial ECM capable of efficient growth factor binding and release, and demonstrated its therapeutic effects on subacute-chronic phase brain stroke, a major unmet medical need. We developed a self-assembling peptide with a bioinspired jigsaw-shaped hydrophobic surface that forms a dovetail-packed superstructure. The jigsaw-peptide formed several-micrometer-long supramolecular nanofibers through an helix-to-strand conformational transition to afford a hydrogel under physiological conditions. The supramolecular hydrogel showed the anomalous rheological properties of an unusually large storage modulus and no weak strain overshoot, indicating robust fiber formation. Furthermore, the nanofibers were three-dimensionally homogeneously dispersed in the hydrogel. Due to these supramolecular properties at the molecular- and macro-scale, the jigsaw-peptide hydrogel could incorporate and release full-length vascular endothelial growth factor (VEGF) efficiently. Single injection of hydrogel incorporating VEGF into injured mouse brain promoted angiogenesis, suppressed neuron death, and improved motor function of subacute-chronic phase brain stroke model mice. This research highlights a therapeutic strategy for injured tissue regeneration using JigSAP supramolecular hydrogel. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-296
Development of 3D culture scaffolds for directional neuronal growth
*Terenzio Marco(1)、Agrawal Lokesh(1)、Guillaud Laurent (1)、Saidani Menouer(1)
*Marco Terenzio(1), Lokesh Agrawal(1), Laurent Guillaud(1), Menouer Saidani(1)
1. Okinawa Institute of Science and Technology Graduate University
Keyword: Neural tissue engineering, Aligned scaffold, Spinal cord and peripheral nerve injury, Laser lithography
Clinical treatment of severe traumatic injuries of the nervous system, a compelling medical urgency, is limited by the complexity of restoring function to the damaged tissue. We study the underlying mechanisms behind peripheral nerve injury and develop systems to promote neuronal regeneration after traumatic injury. In particular, we aim to deploy fibrous 3D scaffold/implants for the directed growth of axons, to restore and repair lost neuronal connections. . However, the fabrication of ideal scaffolds with precisely controlled thickness, texture, porosity, alignment, and with the required mechanical strength,features needed for effective clinical applications, remains technically challenging. We took advantage of state-of-the-art 2-photon photolithography to fabricate highly ordered and biocompatible 3D nanogrid structures to enhance neuronal directional growth. We characterized the physical and chemical properties and proved the biocompatibility of said scaffolds by successfully culturing primary sensory and motor neurons on their surface. Interestingly, axons could grow in 3D between different layers of the scaffold. In addition, axons extended along the fibers with a high degree of alignment to the pattern of the nanogrid, as opposed to the lack of directionality observed on flat glass or polymeric surfaces. Thus, our findings provide a proof of concept and explore the possibility of deploying aligned fibrous 3D scaffold/implants for the directed growth of axons, and could be used in the design of scaffolds targeted towards the restoration and repair of damaged nervous tissue. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-297
Metformin enhances the efficacy of epidermal neural crest stem cells in neuropathic pain
*Afsaneh Asgari Taei(1), Pariya Khodabakhsh(2), Hamed Shafaroodi(3), Leila Dargahi(4)
1. Shahid Beheshti University of Medical Sciences, Tehran, Iran, 2. Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran, 3. Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran, 4. Neurobiology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Keyword: Cognitive deficit, EPI-NCSCs, Metformin, Neuropathic pain
The chemotherapy-induced peripheral neuropathy (CIPN) is a common and challenging complication of cancer treatment which has no efficacious drugs. In recent years, the protective potential of metformin against CIPN has been confirmed in several studies. On the other hand, epidermal neural crest stem cells (EPI-NCSCs) based on autologous accessibility, neural differentiation potency, and high capacity in secreting neurotrophic factors present an exciting potential therapy for neurological disorders. Here, we aimed at evaluating the effect of EPI-NCSCs-treated with metformin on experimental CIPN. Neuropathic pain was induced in male Wistar rats by intraperitoneal injection of paclitaxel eight times every other day. The EPI-NCSCs were treated with the appropriate dose of metformin for 7 days and the treated cells were administered nasally at 48h and 96h after the last dose of paclitaxel. The pain behaviors were determined by von Frey and hot plate four times weekly. Furthermore, cognitive function was assessed by Morris water maze on fifth week. Our findings demonstrate that intranasal injection of metformin-treated EPI-NCSCs leads to relief of mechanical allodynia. Although metformin-treated EPI-NCSCs could not reverse spatial learning performance, but they improved memory deficit compared to control groups. Our data suggest that metformin might enhance the efficacy of EPI-NCSCs in cell transplantation strategies to improve the quality of life for patients suffering from neurotoxicity of cancer treatment. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-298
シポニモドのアストロサイトに対する抗炎症作用
Anti-inflammatory effects of siponimod on astrocyte
*小笠原 陽大(1)、竹内 英之(1)、古宮 裕泰(1)、小川 有紀(1)、高橋 慶太(1)、土井 宏(1)、田中 章景(1)
1. 横浜市立大学 神経内科学・脳卒中医学
*Akihiro Ogasawara(1), Hideyuki Takeuchi(1), Hiroyasu Komiya(1), Yuki Ogawa(1), Keita Takahashi(1), Hiroshi Doi(1), Fumiaki Tanaka(1)
1. Department of Neurology and Stroke Medicine, Yokohama City University
Keyword: siponimod, astrocyte, sphingosine-1-phosphate receptor 1, nuclear factor-kappa B
Objectives: Siponimod is the first oral disease-modifying drug approved for active secondary progressive multiple sclerosis. It acts as a functional antagonist of sphingosine-1-phosphate (S1P) receptor 1 (S1P1) via S1P1 internalization and an agonist of S1P receptor 5 (S1P5). Although S1P1-expressing lymphocyte is the primary target of siponimod in terms of prevention of lymphocytic infiltration into the central nervous system, it is uncertain whether siponimod directly affects astrocytes which express S1P receptors. Here, we investigated the effects of siponimod on astrocytes using mouse primary culture. Methods: Mouse primary astrocyte-enriched cultures were isolated from mixed glial cell cultures prepared from newborn C57BL/6 mouse brains. Astrocytes were activated by stimulation with 1 µg/ml lipopolysaccharide (LPS) or 20 µM hydrogen peroxide for 24 h following pretreatment with 0–1,000 nM siponimod 1 h prior. Astrocytes were pretreated with 1 µM W146 (S1P1 antagonist) or 10 µM suramin (S1P5 antagonist) before siponimod administrations. mRNA expression levels of cytokines, neurotrophic factors, and antioxidants were examined by quantitative PCR. Protein production levels of cytokines were evaluated by ELISA. Activation of nuclear factor-kappa B (NF-κB) was assessed by nuclear translocation of NF-κB using immunostaining. Also, histone deacetylase (HDAC) activity was measured by a cell-based assay. Results: Siponimod significantly suppressed mRNA and protein levels of pro-inflammatory cytokines such as interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) in activated astrocytes. By contrast, siponimod did not affect the expression levels of neurotrophic factors and antioxidants in activated astrocytes. W146 interfered with the suppressive effects of siponimod on astroglial production of IL-6 and TNF-α, but not IL-1β. In addition, treatment with W146 per se reduced activated astroglial production of IL-1β and IL-6, but not TNF-α. Treatment with suramin did not alter astroglial production of these pro-inflammatory cytokines. Both siponimod and W146 suppressed NF-κB activation, whereas only siponimod inhibited HDAC activity. Conclusions: Our findings indicated that siponimod shows broader anti-inflammatory effects on astrocytes than S1P1 antagonists by inhibiting not only NF-κB activation but also HDAC activity. Siponimod might suppress disease progression of multiple sclerosis in part by direct inhibition of astroglial neuroinflammation. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-299
Targeted orexin 1 receptors within the vlPAG for modulation migraine-induced trigeminal nociception, and migraine-related cognitive deficiency in rats
*Kooshki Razieh(1)、Abbasnejad Mehdi (2)
*Razieh Kooshki(1), Mehdi Abbasnejad(2)
1. Department of Biology, Faculty of Sciences, Lorestan University, Khorramabad, Iran, 2. Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran
Keyword: Migraine, Cognition, Learning and memory, vlPAG
Introduction: This study explored whether orexin 1 receptors (Orx1R) in the ventrolateral periaqueductal gray matter (vlPAG) play a role in the modulation of migraine pain in adult male Wistar rats. In addition, migraine-induced subsequent alteration in learning and memory competence was evaluated in migraineurs rats. Methods: To model chronic migraine-associated headache, nitroglycerin (NTG) (5 mg/kg/IP) was administered to test subjects every second day for 9 days. After the last NTG administration, different experimental groups were received intra-vlPAG administration of orexin A (Orx A) or Orx1R antagonist SB334867. Migraine-associated behavioral symptoms were valued in all animals for up to 90 min. Calcitonin gene-related peptide (CGRP)-positive cells were detected in the trigeminal nucleus caudalis (Vc) by immunofluorescence staining. Passive avoidance adeptness and spatial learning and memory performances were evaluated using shuttle box and Morris water maze (MWM) tests, respectively. Further, in vivo electrophysiological field recordings were measured in the CA1 region. Results: NTG caused significant freezing behavior, which was prevented by all OrxA doses. Moreover, OrxA (100 pM) could block NTG-induced increases in facial rubbing and decreases in climbing and body grooming. However, the effects of OrxA were significantly blocked by SB-334867 (20 μM). Besides, OrxA (100 pM) decreased NTG-induced CGRP upregulation. Administration of OrxA decreased NTG-induced learning and memory deficits in MWM and shuttle box. Moreover, OrxA attenuated long-term potentiation deficiency in NTG-injected rats. Conclusions: The data indicated that the activation of Orx1Rs in the vlPAG is effective in relieving NTG-induced migraine symptoms mainly by downregulation of CGRP in the Vc of rats. In addition, the data provided an emerging role for the Orx1R within vlPAG in
the modulation of cognitive decline comorbid with migraine in rats | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-300
低アミロイド原性E61Kトランスサイレチン変異で家族性アミロイドポリニューロパチーが起こる：神経変性機序について
A low amyloidogenic E61K transthyretin mutation causes familial amyloid polyneuropathy: the mechanism of neurodegeneration
*村上 龍文(1)、水口 峯之(2)、三五 一憲(3)、刀祢 重信(4)、砂田 芳秀(1)
1. 川崎医科大学、2. 富山大学、3. 東京都医学総合研究所、4. 東京電機大学
*Tatsufumi Murakami(1), Mineykuki Mizuguchi(2), Kazunori Sango(3), Shigenobu Tone(4), Yoshihide Sunada(1)
1. Kawasaki Medical School, 2. Toyama Medical and Pharmaceutical University, 3. Tokyo Metropolitan Institute of Medical Science, 4. Tokyo Denki University
Keyword: AMYLOID, DRG, NEUROPATHY, TRANSTHYRETIN
Patients with transthyretin (TTR)-type familial amyloid polyneuropathy (FAP) typically show sensory dominant polyneuropathy and autonomic neuropathy. However, the molecular pathogenesis of the neuropathy remains uncertain. In the present study, we characterize the features of FAP TTR E61K. This FAP was late-onset, with sensory dominant polyneuropathy, autonomic neuropathy, and cardiac amyloidosis. Interestingly, no amyloid deposits were found in the endoneurium of the four nerve specimens examined. Therefore, we examined the amyloidogenic properties of E61K TTR in vitro. Recombinant wild-type (WT) TTR, V30M TTR, and E61K TTR proteins were incubated at 37°C for 72 h, and amyloid fibril formation was assessed using the thioflavin-T binding assay. Amyloid fibril formation by E61K TTR was less than that by V30M TTR, and similar to that by WT TTR. E61K TTR did not have an inhibitory effect on neurite outgrowth from adult rat DRG neurons, but V30M TTR did. Furthermore, we studied the sural nerve of our patient by immunohistochemistry, TUNEL and electron microscopy. TTR aggregates were not seen in the extracellular spaces of the nerve by immunohistochemistry. A number of apoptotic cells were observed in the endoneurium of the nerve by TUNEL. Chromatin condensation was confirmed in the nucleus of non-myelinating Schwann cells by electron microscopy. Amyloid fibrils were not seen in the nerve. These findings suggest that E61K TTR is low amyloidogenic, in vitro and in vivo. However, TTR aggregates and amyloid fibrils in the DRG may cause sensory impairments in FAP because the DRG has no blood–nerve barrier. Moreover, Schwann cell apoptosis may contribute to the neurodegeneration. Recently, we demonstrated that DRGs are enlarged on lumbar MRI in an asymptomatic TTR E61K carrier. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-301
The associations between affects and eating behavior in young Korean adults
*Jae-Won Jung(1), Kyung-A Lee(1), Na-Hyun Kim(1), Yukiori Goto(2), Young-A Lee(1)
1. Dept Food Sci & Nutr, Daegu Catholic Univ, Gyeongsan, South Korea, 2. Primate Res Inst, Kyoto Univ, Aichi, Japan
Keyword: EATING BEHAVIOR, MOOD FUNCTION, NEURAL MECHANISM, PSYCHIATRIC DISORDER
Abnormal eating behavior is also quite often associated with several psychiatric disorder, such as drug addiction and mood disorder, suggesting that brain function may have tight relationships with eating behavior. However, such relationships have yet remained largely unclear. In this study, we investigated whether eating behavior is associated with affects in young Korean adults. Fifty (25 female and 25 male) healthy adults aged between 20 and 30 years old participated in the study. A series of questionnaire surveys were administered to assess affects and eating behavior of the participants. Trait and state conditions of affects were evaluated with the Buss-Perry Aggression Questionnaire, Autism-Spectrum Quotient test, 21-item version of the Depression Anxiety Stress Scale, and Ten Item Personality Inventory. Eating behavior was evaluated with the Adult Eating Behavior questionnaire, which addressed 8 dimensions of eating behavior; Enjoyment of Food (EF), Emotional Over-Eating (EOE), Emotional Under-Eating (EUE), Food Fussiness (FF), Food Responsiveness (FR), Hunger (H), Slowness in Eating (SE) and Satiety Responsiveness (SR). Negative affects, such as depression, anxiety, and stress were significantly associated with FR, H, EUE and SR. Affective traits, such as neurotic personality was associated with FR and H, whereas openness was associated with EOE and SR. Moreover, verbal aggression was associated with EUE and SR. These results suggest that some aspects of mood function are associated with eating behavior, and therefore the neural mechanisms linking between moods and eating behavior may be promising candidates for understanding psychiatric disorder. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-302
脊髄損傷後マウスの損傷部位ミクログリアの変化と錐体路神経活動及び運動機能回復過程との関係
Microglia – pyramidal tract interaction in spinal cord injury and their contribution on neural activity and motor function recovery
*大石 遼太郎(1)、山内 一平(1)、中島 宏彰(1)、加藤 大輔(1)、今釜 史郎(1)、和氣 弘明(1)
1. 名古屋大学大学院医学系研究科
*Ryotaro Oishi(1), Ippei Yamauchi(1), Hiroaki Nakashima(1), Daisuke Kato(1), Shiro Imagama(1), Hiroaki Wake(1)
1. Grad Sch Med, Univ of Nagoya, Nagoya, Japan
Keyword: Spinal Cord Injury, Microglia, Neural Actibity
Spinal cord injury (SCI) is a severe pathology that leads to impairment in motor or sensory function. Currently, few effective treatments can be applied for SCI patients and much of the research effort is required to improve neurological outcome. The central area of SCI was consisted by dead cell that surrounded by glial scar that inhibit axonal regeneration. Microglia, one of the glial cell and sole immune cell in Central Nervous System, contribute the immune response to form glial scar after SCI with astrocyte to separate injured lesion and intact lesion. However, the detail of the interaction between microglia and pyramidal tract in the lesion and neural activity change in pyramidal neuron, their relationship with motor functions is remained unclear. To study this issue, we visualized the relation of microglia and pyramidal tract close to the lesion in SCI mice in vivo that may contribute on neural activity change in primary motor cortex using two photon microscope. To visualize microglia and pyramidal tract, we injected an adeno-associated virus (AAV) carrying the tdTomato gene into the layer 5 of left primary motor cortex in CX3CR1-EGFP mice that express GFP specifically in microglia and macrophage. Four weeks after AAV injection, spinal cord was exposed at the apex vertebra with laminectomy, and in SCI mice group, contusion injury was induced on the exposed spinal cord using the IH Impactor. 2,4,7 days after sham/SCI operation, the mice was fixed their spine with fixation device and we observed the interactions between axon and microglia using in vivo two photon microscope.In addition, we injected an AAV carrying the GCaMP gene into the layer 5 of left primary motor cortex in C57BL/6 mice to visualize neural activity in pyramidal tract. Four weeks after AAV injection, we made a sham/SCI operation. And 2,4,7 days after operation, we observed the Ca2+ activity in neuron using two photon microscope. We also applied transcranial direct current stimulation to promote neuronal activity to both sham and SCI group. BMS scores and BBB scores were used for motor function assessment.As previously reported, near the lesion zone after SCI, we observed the number of microglia increased and cell body area extended, which might reflect activation of microglia. We also observed microglia changing their form to rod-shaped ones, which were similar to the microglial change in the brain cortex after brain injury. The process of microglia got polarity along the direction of axon after SCI and it seemed surveying damaged axonal condition moving their processes. Now we are trying to assess whether neural activity affects the microglial features and whether it defines the recovery of motor function. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-303
C6半切マウスにおける、Ia線維の神経活動抑制による痙縮軽減および脊髄神経回路の可塑的変化について
Reduction of Spasticity and Plastic Changes in Spinal Neural Circuits by Inhibition of Ia Fiber activity in C6 Hemi-section Mice
*花﨑 拓人(1)、李 佐知子(1)
1. 名古屋大学医学系研究科
*Takuto Hanasaki(1), Sachiko Lee(1)
1. Grad Sch Med, Univ of Nagoya, Aichi, Japan
Keyword: Spasticity, PLD-mGluR
Introduction: Spasticity is one of the motor dysfunctions that arising from central nerve injury, such as spinal cord injury (SCI). It is recognized as a factor that prevent motor functional recovery. The current treatments are aimed at reduction of muscle tone that causes pain and joint contracture. However, these treatments are not expected to improve radically spasticity and functional recovery. Therefore, it is required to establish new treatment method which is based on the viewpoint of rehabilitation therapy aiming at functional recovery. So, we focused on the afferent type Ia fibers, which are sensory nerves in the spinal stretch reflex arch that caused spasticity. After spinal cord injury, it has been reported that axonal sprouting Ia fibers projecting to a motor neuron increases in excess during hyper stretch reflex. The afferent type Ia fiber neuron is one of proprioceptive sensory neurons that project to the muscle spindle sense changes in muscle length and transmit its action potential afferently to neurons in the spinal cord. There is some stretch-activated channels and voltage-dependent channels in the sensory-acceptor of the muscle spindle. Recently, the metabotropic glutamate receptors (PLD-mGluRs) revealed to exist at this sensory-acceptor and this receptor has functions to enhance and maintain the Ia fiber activity. Dihydroxyphenylglycine (DHPG) has been reported to act as a competitive antagonist for PLD-mGluRs in ex vivo and in vitro of previous studies. Therefore, we hypothesized that moderate suppression of the neural activity of Ia fibers by blocking the PLD-mGluRs with DHPG lead to reduce spasticity. In this study, we investigated to confirm whether DHPG injected to spastic muscle normalize the rate dependent depressions (RDDs) of Hoffman’s reflex (H reflex), that is one of measurement of spasticity, in a mouse model of SCI. Method: C57B/6J female mice were made at semi-transection of 6th cervical spinal cord under deep anesthetize. To test whether an affected muscle exhibited spastic symptoms after SCI, H-reflex Rate Dependent Depression (RDD) was confirmed 1, 2, 3, 4 weeks after SCI. An osmotic pump was implanted in the back of mice, and 10 mM DHPG or PBS as vehicle were administrated to right abductor digiti minimi muscle by tube. Result: One week after surgery, the mice, that were confirmed spasticity with RDDs, were administrated with PBS or DHPG. In the DHPG group, the H reflex was significantly attenuated in a frequency-dependent manner compared with the pre-treatment and PBS groups at 1 week after treatment. Dissection: The results of this experiment tended that the mice of DHPG groups may be normalized in RDDs of H-reflex. Now, we investigate also other time schedule and check the neural plasticity confirming the changes in the number of connections between Ia fiber endings (anti-vGluT1 positive) and a motor neuron (anti-ChAT positive) | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-304
Potentiation of anti-glioblastoma effect with combined interferon-gamma and temozolomide: Role of inflammatory and redox system
*Bashiri Hamideh(1)、Moslemizadeh Amirhossein (2)、Kheirandish Reza(2)、Mohammad Hadi Nematollahi Mohammad Hadi (1)
*Hamideh Bashiri(1), Amirhossein Moslemizadeh(2), Reza Kheirandish(2), Mohammad Hadi Mohammad Hadi Nematollahi(1)
1. Kerman University of Medical Sciences, 2. Faculty of Veterinary Medicine, Shahid Bahonar University of Kerman, Kerman, Iran
Keyword: Glioblastoma, Temozolomide, Interferon Gamma, Toll like Receptor 4
Background: This study aimed to assess the effects of single or combined administration of temozolomide (TMZ) and interferon-gamma (IFN-g) on anxiety-like behaviors, balance disorders, learning and memory, TNF-a, IL-10, some oxidant and antioxidants factors with investigating the toll-like receptor-4 (TLR4) and p-CREB signaling pathway in C6-induced glioblastoma of rats. Methods: In the current study, forty male sprague-dawley rats bearing intra-caudate nucleus (CN) C6 inoculation were randomly divided into five groups as follows: Sham, Tumor, TMZ, IFN-g and a TMZ+IFN-g combination. The open field test (OFT), elevated plus maze (EPM), rotarod and passive avoidance test (PAT) were done on days of 14-17. On day 17 after tumor implantation, rats were sacrificed, brain tissues were extracted for histopathological evaluation. TNF-a, IL-10, SOD, GPX, TAC, MDA were measured by ELISA assay in brain tissue and serum. The expression of the protein level of TLR4 and p-CREB in the brain was also examined by western blot. Results: We found that combination therapy dramatically inhibited the growth of tumor. TMZ, IFN-g and TMZ+IFN-g alleviated the decline of %OAT and %OAE induced by C6. Grooming increased and time spent in center decreased by glioblastoma. IFN-g and TMZ+IFN-g groups amended some C6-induced disturbances in the OFT. The rotarod and PAT demonstrated that treatment groups significantly improved imbalance and memory impairment affected by C6 glioma. SOD, GPX and TAC significantly decreased in tumor group in both serum and brain. TMZ, IFN-g and TMZ+IFN-g groups markedly increased GPX and TAC. The combination group could augment GPX and TAC. In addition, MDA in serum and brain decreased in treatment groups. Our results indicated that TNF-a and IL-10 levels increased significantly in tumor group. Administration of TMZ, IFN-g could reduce TNF-a and IL-10 levels. The combination group declined TNF-a level in serum and IL-10 level in serum and brain. We also found that glioblastoma induced significant upregulation of TLR4 and p-CREB in the brain and combination therapy significantly inhibited the upregulation of TLR4 and p-CREB. Conclusion: The beneficial effects of single and combination therapy on neurocognitive functioning of rats with C6-induced glioblastoma may be mediated via modulating oxidative stress, reduced cytokines, and the downregulation of expression of TLR4 and p-CREB. Combination treatment appears to be more effective than treatment alone. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-305
LINE-1転移モニターマウスを利用した神経系における新規転移解析
Analysis of LINE-1 retrotransposition in the brain with newly created retrotransposition-monitor mice
*藤飯 愼也(1)、文東 美紀(1)、仲地 ゆたか(1)、荒木 喜美(2)、岩本 和也(1)
1. 熊本大学大学院生命科学研究部 分子脳科学講座、2. 熊本大学 生命資源研究・支援センター 疾患モデル分野
*Shinya Fujii(1), Miki Bundo(1), Yutaka Nakachi(1), Kimi Araki(2), Kazuya Iwamoto(1)
1. Department of Molecular Brain Science, Graduate School of Medical Sciences, Kumamoto University, 2. Division of Developmental Genetics, Institute of Resource Development and Analysis, Kumamoto University
Keyword: LINE-1, retrotransposon, L1EGFP, psychiatric disorder
The non-LTR retrotransposon, LINE-1 (long interspersed nuclear element-1, L1) constitutes about 20 % of mammalian genomes. Among many copies of L1, some are still active and can amplify their copies in the host genome by the process called retrotransposition. L1 is the only transposon which can retrotranspose itself using its own proteins. Although retrotransposition is strictly repressed by the host-defense mechanism, it has been reported the somatic L1 retrotransposition in the early neural development and aged brains. We previously reported the increased L1 copy number in the brains of patients with psychiatric disorders and animal models of schizophrenia (Bundo et al., Neuron 2014). To investigate the roles of L1 retrotransposition in the brain in detail, we developed a transgenic mouse line (L1EGFP) in which the retrotransposition can be monitored by the EGFP expression, according to the previous report (Muotri et al., Nature 2010) with some modifications. To avoid the irreversible silencing of L1EGFP construct during passage, we inserted a single copy of the construct at the ROSA26 locus. Currently, we did not see the irreversible silencing of the construct in up to 3rd generation of the new transgenic line. For another improvement, we used EF1α promoter to drive EGFP expression instead of conventional CMV promoter for stable labeling of cells. Interestingly, in contrast to the previous study, the EGFP-positive cells were observed not only in the neuronal cells but also glial cells such as astrocytes, suggesting that retrotransposition affects functions of a wide-range of brain cells. Our mouse will be a valuable tool for studying the physiological and pathophysiological roles of L1 retrotransposition, enabling cell type-specific mapping of retrotransposition in the brain. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-306
日本マウスクリニックにおける網羅的表現型解析
Introduction of comprehensive phenotyping analysis at Japan Mouse Clinic
*山田 郁子(1)、三浦 郁生(1)、澁谷 仁寿(1)、古瀬 民生(1)、田村 勝(1)
1. 理化学研究所バイオリソース研究センター
*Ikuko Yamada(1), Ikuo Miura(1), Hirotoshi Shibuya(1), Tamio Furuse(1), Masaru Tamura(1)
1. RIKEN BioResource Research Center
Keyword: mouse phenotyping, comprehensive analysis, behavioral analysis
In RIKEN BioResource Research Center (BRC), we conduct a comprehensive phenotype analysis project for mutant mice with the aim of better understanding human disease and elucidating gene functions. For this mission, we constructed a systematic phenotyping platform that analyzes more than 700 items, and conducted phenotype analysis of mouse resources such as the genetically modified mice deposited at the BRC. Moreover, we provide the phenotyping platforms to the domestic research community. That is the Japan Mouse Clinic (JMC).
The phenotyping platform of JMC have two pipelines, Pipeline 1 and Pipeline 2. The Pipeline 1 is constructed with comprehensive test items, which consisted visible phenotypic analysis called as RIKEN modified-SHIRPA (8w), hematological analysis (9w), urinalysis (10w), blood chemistry analysis (11w), intraperitoneal glucose tolerance test (14w), adipocytokine analysis (18w), fundoscopic analysis (19w), bone morphology analysis using X-ray (22w), body composition analysis by dual-energy X-ray absorptiometry (22w), electrocardiogram analysis (25w), immunophenotyping by fluorescence-activated cell sorting (26w) and autopsy and histological analysis (26w). These tests are compliant with international standardized phenotyping protocols, which are used by International Mouse Phenotyping Consortium (IMPC) project, and are accompanied by metadata.
The Pipeline 2 specialized in behavioral analysis, which can detect psychiatry-relevant behavioral abnormalities, especially autistic spectrum disorder and emotional abnormality. This behavioral pipeline is conducted in order of following, light/dark transition test (anxiety: 6w), open-field test (spontaneous activity: 7w), Crawley’s social interaction test (social behavior: 9w), home-cage activity test (activity pattern: 10-11w), Y-maze test (persistence and spatial memory: 12w), fear conditioning test (learning and memory: 13w), and pre-pulse inhibition test (sensory gating: 14w).
In this presentation, we will introduce JMC phenotyping pipelines while showing the actual data of mice. The JMC found new phenotypes in genetically modified mice and we expect it to be re-evaluated as a new human disease model. Furthermore, we will correlate clinical data of human disease with mouse phenotype information to further enhance the value of mouse resources in Japan and contribute to its development and intellectual infrastructure.
JMC: http://mouseclinic.brc.riken.jp/ | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-307
マウス新生仔の鼻腔炎症による腸内細菌叢の変動と脳組織損傷
Perturbation of gut microbiota and brain tissue damages caused by neonatal nasal inflammation
*石井 さなえ(1)、浅野 妃南(2)、三島 由祐子(1)、大﨑 敬子(3)
1. 杏林大学保健学部、2. 杏林大学大学院保健学研究科、3. 杏林大学医学部感染症学
*Sanae Hasegawa-Ishii(1), Hinami Asano(2), Yuko Mishima(1), Takako Osaki(3)
1. Fac Health Sci, Kyorin Univ, 2. Grad Sch Health Sci, Kyorin Univ, 3. Dept Infect Dis, Sch Med, Kyorin Univ
Keyword: Nasal inflammation, Microbiota, Gut-Brain axis
Maternal infection and resulting immune activation are risk factors for neuropsychiatric disorders in offspring. But the mechanisms underlying sustained impairment of the brain function after birth remain to be elucidated. We hypothesized that inflammation during fetal and neonatal periods may disturb the gut microbiota and disrupt the gut-brain communication, leading to sustained brain dysfunction. To examine this hypothesis and the sex differences in the response to the inflammation, neonatal male and female mice received bilateral nasal administration of 10 μL/nostril of lipopolysaccharide (LPS) as a nasal inflammation model, or saline as a control, from postnatal day 7 (P7) twice a week for 3 weeks (at P7, P10, P14, P17, P21 and P24). After P24, mice were weaned and housed in groups for male or female separately. Body weight was measured once a week. At P24 and 10 weeks (10wks) of age, feces were collected for 16SrRNA metagenome analyses. At the same time points, mice were fixed and frozen sections were made for histological analysis. Results indicated that the body weight was similar in saline- and LPS-treated mice until P24 in both male and female, but that it was significantly lower in LPS-treated male mice than in saline-treated male mice after P24. The gut microbiota was perturbed in the phylum, family and genus levels in LPS-treated male mice at P24, while it did not change in LPS-treated female mice compared to saline-treated control. At 10wks, the abundance of a few bacteria was altered in LPS-treated male mice, but not in LPS-treated female mice compared to saline-treated control. Histological analyses were now ongoing. These results suggest that neonatal nasal inflammation causes long-term perturbation of gut microbiota and that male mice seemed to be more affected. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-308
ジフェニルアルシン酸による培養ラット小脳由来アストロサイトにおけるNF-κBの活性化と異常活性化
Diphenylarsinic acid-induced activation of NF-κB signaling pathways in cultured rat cerebellar astrocytes
*根岸 隆之(1,2)、梶田 直博(1)、佐々木 翔斗(2)、都築 孝允(1)、湯川 和典(1,2)
1. 名城大学薬学部、2. 名城大学大学院薬学研究科
*Takayuki Negishi(1,2), Naohiro Kajita(1), Shoto Sasaki(2), Takamasa Tsuzuki(1), Kazunori Yukawa(1,2)
1. Faculty of Pharmacy, Meijo University, 2. Graduate School of Pharmacy, Meijo University
Keyword: Astrocytes, Arsenic, MAP kinase, Oxidative stress
Diphenylarsinic acid (DPAA) was one of non-natural pentavalent arsenic compounds found in the well water in Kamisu, Ibaraki, Japan in 2003, which induced neurological symptoms including cerebellar symptom in people having used this well water for domestic use. Our previous studies revealed that DPAA could induced the aberrant activation of cultured rat cerebellar astrocytes, which included promotion of cell growth, increases in phosphorylation of major mitogen-activated protein kinases (MAPKs) (ERK1/2, p38MAPK, and SAPK/JNK), activation of downstream signaling pathways such as transcription factors (CREB, c-Jun, and c-Fos) and antioxidant proteins (Nrf2, heme oxygenase-1 (HO-1), and heatshock protein 70 (Hsp70)), and induction of glutathione secretion. NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) was known to play a key role in oxidative stress-related neurodegenerative diseases. Here, we investigated a possible role of NF-κB in DPAA-induced aberrant astrocyte activation using an inhibitor of NF-κB activation, sulfasalazine (SSZ). Cultured rat cerebellar astrocytes were prepared from P2 Wister rats, maintained in DMEM/F-12/10%FBS, and exposed to DPAA (0 or 10 µM) and SSZ (0, 20, or 40 µM) for 96 h in serum-free condition (DMEM/F-12/ITS-X). DPAA exposure increased phosphorylation of NF-κB p65 subunit but not its expression and stimulated its nuclear translocation. SSZ counteracted DPAA-induced expression of Nrf2, HO-1, and Hsp70. SSZ suppressed DPAA-induced increases in phosphorylation of p38MAPK and SAPK/JNK but not ERK1/2. SSZ ameliorated DPAA-induced expression of c-Jun and c-Fos and little affected DPAA-induced CREB phosphorylation. Furthermore, SSZ suppressed DPAA-induced cell growth and glutathione secretion. These results suggested that DPAA could induce NF-κB activation (phosphorylation and nuclear translocation), which adversely contributed to DPAA-induced aberrant astrocyte activation partially. NF-κB suppression by SSZ might be a possible therapeutic strategy for DPAA poisoning, although the potential effect of SSZ as the cystine-glutamate antiporter, system xc-, should be considered additionally in the future study. | | 2022年6月30日 13:00～14:00 宜野湾市民体育館 ポスター会場2
1P-309
多施設データにおける安静時機能的結合を用いた摂食障害分類
Resting-state functional connectivity MRI classification of eating disorder in multi-site data
*高村 恒人(1)、佐藤 康弘(2)、平野 好幸(3)、磯部 昌憲(4)、戸瀨 景茉(4)、野田 智美(4)、兒玉 直樹(5)、吉原 一文(6)、守口 善也(1)、関口 敦(1)
1. 国立精神・神経医療研究センター, 精神保健研究所, 行動医学研究部、2. 東北大学病院心療内科、3. 千葉大学 子どものこころの発達教育研究センター、4. 京都大学大学院 医学研究科 脳病態生理学講座 精神医学教室、5. 産業医科大学医学部 神経内科学講座、6. 九州大学大学院医学研究院 心身医学
*Tsunehiko Takamura(1), Sato Yasuhiro(2), Yoshiyuki Hirano(3), Masanori Isobe(4), Keima Tose(4), Tomomi Noda(4), Naoki Kodama(5), Kazufumi Yoshihara(6), Yoshiya Moriguchi(1), Astushi Sekiguchi(1)
1. Department of Behavioral Medicine, National Institute of Mental Health, National Center of Neurology and Psychiatry, Tokyo, Japan, 2. Department of Psychosomatic Medicine, Tohoku University Hospital, Miyagi, Japan, 3. Research Center for Child Mental Development, Chiba University, Chiba, Japan, 4. Department of Psychiatry, Graduate School of Medicine, Kyoto University, Kyoto, Japan, 5. Division of Psychosomatic Medicine, Department of Neurology, University of Occupational and Environmental Health, Kitakyushu, Japan., 6. Department of Psychosomatic Medicine, Kyushu University, Fukuoka, Japan
Keyword: Resting-state functional connectivity, Eating disorder, multi-site data
The resting-state functional connectivity MRI (rsfcMRI) has attracted attention as a tool to provide biological indicator helping with diagnosis of neuropsychiatric disorders such as eating disorder (ED). RsfcMRI in combination with machine learning is suggested to be able to discriminate between a healthy participant and a patient with ED. To reliably apply machine learning to thousands of connectivity from rsfcMRI, we need to have access to big data, which can be most efficiently acquired from multiple clinical centers. A problem in a multiple center study, however, is that it involves different MRI scanners often from different vendors. This poses substantial and systematic differences in quality in the dataset for machine learning, but no established method is available to handle with differences in image quality across MRI scanners. To solve this issue, we applied a method to reduce differences in MRI data quality across study sites, using a combat. We used rsfcMRI data from 120 healthy volunteers and 115 patients with ED who were scanned at 5 different sites with 3 types of MRI scanner. After preprocessing, we retrieved functional connectivity (FC) from rsfcMRI, using Automated Anatomical Labeling (AAL). After applying ComBat to the generated multidimensional vectors, dimensionality reduction was performed. We then used a linear support vector machine classifier, which was trained to find a hyper-plane for segregating the patterns of FCs in the patients with ED from those of the healthy volunteers. The linear support vector machine classifier successfully classified the two groups, with accuracy of about 70%. The present findings support the utility of ComBat as a tool for dimensional reduction and for reducing site differences for multi-center rsfcMRI to before feeding rsfcMRI data into a machine learning based-classifier as a subsidiary diagnostic tool of ED. | |
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