Poster

Poster 11 Memory, Emotion, Behavior 1 ポスター 11 記憶、情動、行動1

P11-1 The pivotal role of PACAP-induced astrocyte-neuron lactate shuttle in fear memory acquisition and retrieval PACAP誘発アストロサイト-ニューロン乳酸シャトルは恐怖記憶の獲得および想起へ関与する Kambe Yuki（神戸悠輝）¹，中島優¹，Nguyen Thanh Trung¹，新谷紀人²，橋本均^2,3，吉武尚⁴，Kehr Jan⁴，栗原崇¹，宮田篤郎¹ ¹Dep. Pharmacol., Grad. Sch. Med. Dent. Sci., Kagoshima Univ. ²Lab. Mol. Neuropharmacol., Grad. Sch. Pharmaceut. Sci., Osaka Univ. ³United Grad. Sch. Child Dev., Osaka Univ. ⁴Lab. Pharmacol. Neurochem., Dept. Physi. Pharmacol., Karolinska Institutet Previously, it reported that PACAP/PAC1 receptor signaling is important for fear memory acquisition, however the signaling mechanism still remains largely unclear. Since astrocytic glycogenolysis and lactate secretion are recently shown to be essential for fear memory, we aimed to elucidate a possible involvement of astrocyte-neuron lactate shuttle (ANLS) in the development of PACAP-induced fear memory acquisition and retrieval. While the glycogen amount of cultured forebrain astrocytes was dose-dependently decreased by PACAP, the lactate concentration in culture supernatant was significantly increased by the PACAP exposure, suggesting that PACAP induced ANLS activation in the cultured forebrain astrocytes. In vivo system, PACAP (-/-) mice is known to acquire less fear memory in passive avoidance and contexture- and cue- mediated fear conditioning test than in PACAP (+/+) mice. When fear memory was acquired, glycogen amount was significantly decreased, while lactate amount was significantly increased in the hippocampus of +/+ mice, but not in -/- mice. On the other hand, when fear memory was retrieved, glycogen contents in the amygdala and hippocampus of +/+ mice were decreased, and lactate contents in the medial prefrontal cortex and amygdala were increased in +/+ mice, but the changes in the medial prefrontal cortex and amygdala were not observed in -/- mice. In vivo microdialysis study revealed that intracerebroventricular injection of PACAP induced prolonged increase in the extracellular lactate level in the hippocampus of wild type rats. These results suggested that PACAP-mediated ANLS activation is important for fear memory acquisition and retrieval. Future study would elucidate intracellular signaling mechanisms how PACAP evokes ANLS.		P11-2 The analysis of neuropsin dependent and independent late associative memory ニューロプシン依存的・非依存的長期連合性記憶の解析 Ishikawa Yasuyuki（石川保幸），鈴木結花，依田祐也 Department of Systems and Life Engineering, Maebashi Institute of Technology, Gunma, Japan Synaptic plasticity is widely accepted to provide a cellular basis for learning and memory. Synaptic associativity could be involved in activity-dependent synaptic plasticity, because it distinguishes between local mechanisms of synaptic tags and cell-wide mechanisms that are responsible for the synthesis of plasticity-related proteins. An attractive hypothesis for synapse specificity of long-term memory (LTM) is synaptic tagging: synaptic activity generates a tag, which captures the plasticity-related proteins derived outside of synapses. Previously we have been reported that neuropsin, a plasticity-related extracellular protease, was involved in synaptic tag setting. In the present study, we tested the hypothesis that neuropsin was engaged in behavioral tagging for LTM in late associative memory. Behaviorally, weak training inhibitory passive avoidance task (IA) or spatial object recognition task (SOR), which induces short-term memory (STM) but not LTM, can be consolidated into LTM by exposing animals to novel but not familiar environment 1 h before training. We found that neuropsin deficient mouse impaired such transformation short-term into long-term memory by exposure to novelty in IA, but not SOR. These results suggest that the presence of neuropsin-dependent and -independent late associative memory.

P11-3 Abnormal neural activation patterns underlying working memory impairment in chronic phencyclidine-treated mice Phencyclidine慢性投与マウスが示すワーキングメモリー障害の神経基盤の探索 Arime Yosefu（有銘預世布），秋山一文 Department of Biological Psychiatry and Neuroscience, Dokkyo Medical University School of Medicine Working memory impairment is a hallmark feature of schizophrenia and the neural circuit anomalies underlying this impairment are poorly understood. The aim of this study is to assess working memory performance in the chronic phencyclidine (PCP) mouse model of schizophrenia, and to identify the neural substrates of working memory. To address this issue, we conducted the following experiments for mice after withdrawal from chronic administration (14 days) of either saline or PCP (10 mg/kg): (1) a discrete paired-trial variable-delay task to assess working memory, and (2) brain-wide c-Fos mapping to identify activated brain regions relevant to this task performance either 90 min or 0 min after the completion of the task, with each time point examined under working memory effort and basal conditions. Correct responses in the test phase of the task were significantly reduced across delays (5, 15, and 30 s) in chronic PCP-treated mice compared with chronic saline-treated controls, suggesting delay-independent impairments in working memory in the PCP group. In layer 2-3 of the prelimbic cortex, the number of working memory effort-elicited c-Fos+ cells was significantly higher in the chronic PCP group than in the chronic saline group. The main effect of working memory effort relative to basal conditions was to induce significantly increased c-Fos+ cells in the other layers of prelimbic cortex and the anterior cingulate and infralimbic cortex regardless of the different chronic regimens. Conversely, this working memory effort had a negative effect in the ventral hippocampus. These results shed light on some putative neural networks relevant to working memory impairments in mice chronically treated with PCP, and emphasize the importance of the layer 2-3 of the prelimbic cortex.		P11-4 Effect of post-reexposure administration of KNT-127 on extinction learning or reconsolidation of contextual fear memory in rats 条件づけ文脈への再曝露後のKNT-127投与が恐怖記憶の消去学習あるいは再固定化に及ぼす影響 Furuie Hiroki（古家宏樹）¹，赤木希衣^1,2，山田美佐¹，斎藤顕宜¹，長瀬博³，岡淳一郎²，山田光彦¹ ¹Dept of Neuropsychopharmacol, NCNP, Tokyo, Japan ²Lab of Pharmacol, Faculty of Pharm Sci, Tokyo Univ of Science, Chiba, Japan ³WPI-IIIS, Univ of Tsukuba, Ibaraki, Japan Recently, we demonstrated that KNT-127, a δ opioid receptor agonist, reduces fear response after extinction when it was administered before reexposure to the training context in rats. We also reported that KNT-127 may attenuate reconsolidation of fear memory if administered before the short reexposure to the context. However, whether KNT-127 truly facilitates extinction learning or attenuates reconsolidation of contextual fear memory is not known. To answer these questions, we undertook experiments on contextual fear conditioning with post-reexposure administration of KNT-127. In conditioning session, male Wistar rats were placed into a conditioning chamber for 180 sec, then three footshocks were delivered. After additional 60 sec, rats were returned to their home cage. Twenty-four-hour after conditioning, rats were reexposed to the conditioning chamber for 0, 3, or 10 min without footshock then they were administered KNT-127 (3 mg/kg) or saline subcutaneously. On the day following reexposure session, rats were placed in the conditioning chamber for 6 min and freezing time was recorded. In this study, regardless of the duration of reexposure, rats administered KNT-127 after the reexposure session showed equivalent freezing in the test session, when compared to the rats injected with saline. It is demonstrated that KNT-127 does not affect extinction learning or reconsolidation of contextual fear memory. Previously, we demonstrated that KNT-127 shows anxiolytic-like effects in behavioral tests for anxiety in rats. Together, it is suggested that robust anxiolytic-like effect of KNT-127 reinforced the new associative learning during the reexposure to the context when administered before the session, and thereby reduced fear response on the following day.

P11-5 Effect of NAD+ supplementation on mice behavior NAD投与によるマウスの行動変化の解析 Gerasimenko Maria¹，Cherepanov Stanislav²，横山茂²，東田陽博² ¹United Graduate School of Child Development, Osaka University, Kanazawa University, Hamamatsu University School of Medicine, Chiba University and University of Fukui ²Kanazawa University Research Center for Child Mental Development NAD+ is one of the essential biomolecules, which participates in a large number of vital processes like ATP synthesis, redox homeostasis, and signal pathways. A lot of studies revealed positive effects of NAD+ precursors’ supplementation on the physical condition in a case of neurodegenerative disorders, muscular dystrophy, and senescence. In some studies was found an anxiolytic effect of elevating the NAD+ level. NAD+ as a substrate in ADP-ribosyl cyclase reactions is one of the crucial components for oxytocin release. Through this pathway elevating NAD+ contents possibly can lead to enhancing of oxytocin release and so can influence on social behavior. However, it has never been intensively studied whether elevation of NAD+ levels can affect behavior, especially in emotional behaviors. To assess this question, we used CD157 knockout mice with a genetic background of C57BL6 - a model which displayed behavioral changes: anxiety, depression and social deficit. CD157KO and C57BL6 mice were treated with an NAD+ precursor at the dose of 3 or 13 mg/mouse/day in PBS solution or placebo in equal volume through gavage. The procedure of gavage was performed daily in the same time period for 12 days. On 13th day behavioral experiments: three-chamber and light-dark transition tests were performed. Additionally, levels of oxytocin in the CSF and NAD+ in the brain tissue were measured. Precursor supplementation significantly elevates an NAD+ content in the brain tissue of both genotypes. The dose of 3 mg significantly elevate the oxytocin level in CSF of CD157 KO mice, but not in the wild-type. Both doses of the precursor attenuate pathological high anxiety and improve the social behavior in CD157KO mice. The elevated NAD+ level had no effect on C57BL6 mice behavior.		P11-6 Analysis of paternal aggression in ICR strain マウスの授乳時の父親の攻撃行動についての解析 Shabalova Anna，りゃんみんくん，東田陽博 Research Center for Child Mental Development, Kanazawa University, Kanazawa, Japan Analysis of paternal aggression in ICR strain mice.The development of healthy offspring needs both maternal and paternal cares. Lactating dams display aggression to protect their pups as a part of parental behavior. However, less information on requirement, hormones and mechanisms is available for sire’s aggression. We analyzed paternal aggressive behavior in different conditions based on the Resident-Intruder Test. After pairmates, the sires were isolated to the new individual cage for 1 week. The sires were divided to three groups according to the different responses of cues. In the first group, sires were housed alone. In the second group, the sires were exposed to their dams daily for 6 hours. In the third group, sires were exposed to their pups daily for 6 hours. Following that, sires were tested for the offensive aggression using the Resident-Intruder Test for 3 consecutive days. On the third day of the behavioral experiment sires were killed one hour after the beginning of the test for analyzing expression of c-fos, oxytocin-immunoreactive cells and the colocalization of c-fos and oxytocin in the paraventricular nucleus (PVN) of the hypothalamus. Sires, exposed to the pups demonstrated the lowest attack latency and the highest number of attacks and duration of aggression, whereas, in contrast, sires spent alone showed the highest latency to attack and the lowest number of aggression and duration of attacks. The results are associated with the number of c-fos in the PVN. Pups-exposure increased c-fos-positive cell number in the PVN and percentage of the colocalization with oxytocin neurons. These finding suggest that paternal aggression is likely regulated by oxytocin in ICR male mice.

P11-7 The new analogs of oxytocin.: Activity, selectivity, and effects on behavior オキシトシンの新規な類似体。: 活性、選択性と行動への効果 Cherepanov Stanislav¹，一ノ瀬亘²，由比光子¹，Shabalova Anna¹，横山茂¹，周東智²，東田陽博¹ ¹Research Center for Child Mental Development, Kanazawa University ²Laboratory of organic chemistry for drug development. Hokkaido University Autism spectrum disorder is a neurodevelopmental disorder characterized by impairments in social communication and repetitive behaviors. A great deal of research has focused on neuroendocrine hormone, oxytocin (OT), for the treatment of the symptom of social deficits. However, OT has a short time of half-life in blood and high affinity to vasopressin receptors. According to this, we newly designed two oxytocin analogs: X1 and X2. As in vitro experiments, we investigated release of intracellular calcium in stable transformed cell lines to express OTR, V1AR or V1BR. X1 increased calcium concentrations with higher Emax than OT and demonstrated a superagonistic profile, while X2 demonstrated partial effects. X1 and X2 had significantly lower activity to V1A and V1B-receptors. Data of ex-vivo evaluation through uterine-contraction measurements indicated ability of both compounds to induce contractions, as agonists. We investigated OT and two analogs on paternal behavior tests with CD38 knockout (CD38KO) mice. CD38KO male mice demonstrate low pup retrieving scores. In the case of OT, X1 or X2, at 30 minutes after injection, CD38KO male mice increased retrieving scores. After 3-24 hours, OT completely lost such effects, X1 and X2 demonstrated higher effects on parental scores. Data of pharmacokinetic studies indicated absence of difference in metabolism of OT and X-analogs in blood, but indicated an ability to induce a long-lasting elevation of blood OT concentrations, suggesting analog-induced OT-release. Overall data indicates that two novel OT analogs have high affinity and activity to OT-receptors, with a high selectivity and possess immediate and long-lasting effects. However, a future study to investigate mechanisms of long-term activity of new compounds is required.