TOP ＞ ポスター

ポスター E. 行動の分子・神経的基盤 1 E. Molecular and Neuronal Bases of Behavior 1 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-011 小鳥のさえずりの内発的動機づけに関わる脳領域の同定 Identification of the brain area associated with intrinsic motivation for singing in zebra finches *森 千紘(1)、Kim Yunbok(2)、青木 直哉(1)、本間 光一(1)、小島 哲(2) 1. 帝京大学、2. 韓国脳研究院 *Chihiro Mori(1), Kim Yunbok(2), Naoya Aoki(1), Koichi Homma(1), Satoshi Kojima(2) 1. Teikyo University, 2. Korea Brain Research Institute Keyword: songbird, motivation, Arc, IEGs Behaviors driven by intrinsic motivation are critical for developing and optimizing cognitive, social, and physical functions throughout life. Intrinsic motivation arises within individuals for internal satisfaction, sharply contrasting with extrinsic motivation, which involves engaging in a behavior to gain external rewards or avoid punishment. [MC1] One major obstacle in understanding the neural mechanisms of intrinsic motivation at the circuit and cellular level is the lack of a suitable animal model. Songbirds, such as zebra finches, offer a unique opportunity to study neural substrates of intrinsic motivation, because they spontaneously produce many renditions of songs with highly-quantifiable structure for vocal practice, even in the absence of apparent recipients (undirected singing). Here, we established a simple behavioral procedure to easily manipulate and quantify intrinsic motivation for undirected singing in adult male zebra finches: Temporary suppression of undirected singing by turning off the ambient light dramatically enhanced intrinsic motivation for singing, and the degree of enhancement depended on the duration of suppression. Moreover, to identify brain areas associated with the enhancement of singing motivation, we explored brain areas showing activated expression of the immediate early genes (Arc, c-fos, or Egr-1) in birds with relatively high singing motivation but not in birds with low singing motivation. We found that Arc mRNA expression in the nucleus of the hippocampal commissure (NHpC) was activated in the birds with high singing motivation, suggesting that neural activity in NHpC is associated with intrinsic singing motivation. In addition, such activation was not observed in NHpC of female birds, which do not produce songs, under the condition that increases singing motivation in male birds, further supporting the involvement of NHpC of male birds in intrinsic singing motivation. These results provide an important first step toward understanding detailed neural mechanisms underlying intrinsic motivation for complex, learned motor behaviors. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-012 報酬追求を継続するための中脳ドーパミン細胞の相反する報酬予測誤差信号 Midbrain dopamine neurons signal opposing reward prediction errors to continue reward pursuit *石野 誠也(1)、鎌田 泰輔(1)、Gideon Sarpong(1)、司 怜央(1)、向平 妃沙(1)、小林 憲太(2)、本田 直樹(3)、大石 直也(1)、小川 正晃(1) 1. 京都大学大学院 医学研究科、2. 生理学研究所 行動・代謝分子解析センター、3. 広島大学大学院 統合生命科学研究科 *Seiya Ishino(1), Taisuke Kamada(1), Sarpong Gideon(1), Reo Tsukasa(1), Hisa Mukohira(1), Kenta Kobayashi(2), Naoki Honda(3), Naoya Oishi(1), Masaaki Ogawa(1) 1. Grad Sch Med, Kyoto University, Kyoto, Japan, 2. Center for Genetic Analysis of Behavior, National Institute for Physiological Sciences, Aichi, Japan, 3. Grad Sch Integrated Sciences for Life, Hiroshima University, Hiroshima, Japan Keyword: DOPAMINE, MOTIVATION In order to obtain an uncertain reward, it is necessary to continuously pursue the reward even when it is unexpectedly omitted. Classic learning theories assume that reward prediction errors (RPE), defined as the discrepancy between actual and expected reward, are negative in that situation, which is supposed to decrease our motivation. Midbrain dopamine neurons are thought to signal RPE, and their activity is inhibited in the face of omissions of the expected reward. However, in order to continuously pursue the reward, a neural substrate that responds in the opposite way to the canonical phasic inhibition of dopamine neurons is necessary. We previously found that rats responded faster in anticipation of uncertain reward than certain reward in an instrumental task, in which different sensory cues were associated with different probabilities of reward (Ogawa et al, Neuron, 2013). This behavior could be explained by assuming that rats increased their motivation in the face of omission of expected reward (Esber & Haselgrove, 2010). To reveal the neural substrates, we developed a behavioral task in head-restrained rats which enabled us to accurately monitor reward-seeking behavior after an uncertain reward is not obtained. We recorded the electrical activity of dopamine neurons in the lateral ventral tegmental area (VTA) and measured dopamine release in the nucleus accumbens (NAc). In the face of omission of expected reward, we found dopamine neurons with increased activity (type 2) that followed decreased activity of canonical RPE-type dopamine neurons (type 1). Further, type 2 neurons also showed inverse responses to type 1 neurons at both reward receipt and reward termination. Moreover, the increased activity of type 2 neurons and the consequent increase in the dopamine release in the NAc correlated with subsequent switching to the next reward-seeking behavior. Optogenetic stimulation of dopaminergic input to the NAc during reward omission increased reward-seeking behavior. Thus, we conclude that type 1 and type 2 dopamine neurons cooperate to regulate continued reward pursuit. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-013 ゼブラフィッシュ意欲行動の神経基盤 A neural substrate for motivated behavior in zebrafish *宮坂 信彦(1)、吉原 良浩(1) 1. 理研CBS システム分子行動学研究チーム *Nobuhiko Miyasaka(1), Yoshihiro Yoshihara(1) 1. Lab. for Systems Molecular Ethology, RIKEN CBS Keyword: motivation, olfactory, learning, novel environment One of the most important roles of olfaction is to learn and memorize odor cues associated with feeding. The learned odor cues allow animals to efficiently find foods and promote eating, which is critical for the survival of most species in natural habitats. However, how learning enables odor cues to evoke food-seeking motivation and the underlying circuit mechanisms remain largely unknown. Here we use a simple behavioral paradigm for appetitive olfactory conditioning in zebrafish and analyze brain regions that are activated after learning. Infusion of a synthetic odorant into a test tank does not elicit obvious behavioral changes in naive zebrafish, whereas repeated pairings of the odorant infusion with feeding result in odorant-evoked attraction to the odorant source and/or food ports prior to feeding. Following the probe trial, the number of c-fos-positive cells in a specific subnucleus of the thalamus is significantly greater in paired group than that in unpaired group, while c-fos+ cells in Dp and Dm (homologue of piriform cortex and pallial amygdala, respectively) do not differ in number between the two groups. We also find that c-fos+ cells increase in number in the thalamic subnucleus upon exposing naive zebrafish to a novel environment. Double in situhybridization analyses reveal that the thalamic c-fos+ cells in both the paired group and the novel environment group are a specific population of excitatory neurons expressing corticotropin-releasing hormone (crh) gene. These results suggest that activation of the crh+ thalamic neurons correlates with a brain state common to motivated behaviors including appetitive behavior evoked by a reinforced odorant and exploratory behavior in a novel environment. Further experiments are ongoing to clarify connectivity and function of the crh+ thalamic neurons. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-014 インポーチンα3欠損マウスの動機づけ異常とその脳内ネットワーク Motivational Abnormalities and Their Brain Networks in Importin α3 deficient Mice *青峰 良淳(1,2)、櫻井 航輝 (1,2)、マクファーソン トム(1,2)、小澤 貴明(1,2)、宮本 洋一(3)、米田 悦啓(4)、岡 正啓(3)、疋田 貴俊(1,2) 1. 大阪大学・蛋白質研究所、2. 大阪大学大学院・理学研究科生物科学専攻、3. 医薬基盤研究所、4. 国立研究開発法人医薬基盤・健康・栄養研究所 *Yoshiatsu Aomine(1,2), Koki Sakurai(1,2), Tom Macpherson(1,2), Takaaki Ozawa(1,2), Yoichi Miyamoto(3), Yoshihiro Yoneda(4), Masahiro Oka(3), Takatoshi Hikida(1,2) 1. Inst. for Protein Res, Osaka Univ, Osaka, Japan, 2. Dept. Biol., Grad. Sch. Sci., Osaka Univ, Osaka, Japan, 3. National Inst.of Biomedical Innovation, Osaka, Japan, 4. National Inst.of Biomedical Innovation, Health and Nutrition (NIBIOHN), Osaka, Japan Keyword: motivation, progressive-ratio schedule, c-Fos, network analysis Importin α3 (Gene: Kpna3, ortholog of human Importin α4) is a subtype of the importin α family, which function in nucleocytoplasmic transport by forming trimeric complexes between cargo proteins and importin β1. Each importin α subtype (6 in mice and 7 in humans) displays different, but partially overlapping, cargo substrate specificities, and subtype specific expressional regulation is known to play a critical role in neuronal differentiation. Importantly, multiple studies on humans have associated single nucleotide polymorphisms (SNP) in KPNA3 with various psychiatric disorders such as schizophrenia, major depression, and substance addiction. However, the regulatory roles of importin α3 regarding brain function and behavior have not been investigated so far. Recently, in-depth analysis on the behavioral characteristics of patients have uncovered abnormalities in reward processing which underlie the widespread behavioral deficits seen in psychiatric disorders. In this study, we evaluated the behavioral effects of Kpna3 knockout (KO) in mice on touchscreen operant chamber-based tasks evaluating reward-seeking motivation (progressive-ratio schedule). Kpna3 KO mice showed a significant increase in reward seeking motivation (break point). We also measured the number of c-Fos-positive cells, a marker of neural activity, in 20 regions of the brain and predicted interregional functional connectivity based on network analysis in a progressive ratio schedule. Network analysis suggests that Kpna3 deficiency increased overall interregional connections. In particular, significant increases in functional connectivity were found in the nucleus accumbens, ventral pallidum, lateral hypothalamus, and amygdala. Analysis of centrality based on graph theory of networks (for detection of hubs) indicated an overall increase in degree centrality of regions. The lateral hypothalamus showed a higher degree centrality and betweenness centrality in KO, despite lower centrality in wild type. Thus, it is suggested that Kpna3 deficiency increases the overall interregional connectivity during motivational behavior. These findings illuminate the importance of KPNA3 in the mechanisms of reward seeking motivation, and that Kpna3 KO mouse may be a potential model for investigating the motivational abnormalities seen in patients with psychiatric disorders. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-015 呼吸への注意が粘り強さと前頭葉活動に及ぼす影響 Effects of focused attention to breathing on perseverance and prefrontal activity *細川 貴之(1)、水子 学(1)、中村 有里(1)、彦坂 和雄(1) 1. 川崎医療福祉大学 *Takayuki Hosokawa(1), Manabu Mizuko(1), Yuri Nakamura(1), Kazuo Hikosaka(1) 1. Kawasaki University of Medical Welfare Keyword: mindfulness, prefrontal cortex, fNIRS Mindfulness meditation (MM) is a form of mental training with focused attention to breathing. It has many positive effects such as stress reduction, and attention improvement. However, the effects of MM on behavior and brain activity are not fully understood yet. Here, we examined the effects of focused attention to breathing on behavioral perseverance when in a difficult situation, where frequent failures could occur. We conducted a “stopwatch task” in an experimental setting where repetitive failures could occur. In this task, subjects were instructed to press a button to start a timer, count seconds in their head, and stop the timer at 10 s with an allowable range of deviation. As the subject succeeded in the trials, the allowable range of deviation was gradually decreased. Thus, the task became increasingly difficult, and the subjects tended to fail in the task. Even when the subjects failed to stop the timer within the allowable range of deviation, they could continue the task as many times as they wanted. They could also discontinue the task any time. The number of how many trials they performed before discontinuing the task was used as the index of their perseverance. We also recorded cortical hemodynamics in the prefrontal cortex (PFC) by functional near-infrared spectroscopy (fNIRS) during the stopwatch task. The subjects’ performance in the first session of the stopwatch task was considered as their baseline performance. We assigned the subjects to either the experimental (BREATH) or control (CTRL) group. After the first session, the BREATH group had one week of MM training by watching a 10-min instruction movie about MM every day at home. One week after the first session, both groups had the second session of the stopwatch task. The BREATH group was instructed to pay attention to their breathing during every intertrial interval. We found significant increases in perseverance and response accuracy (discrepancy of response time from 10 s) in the BREATH group but not in the CTRL group. We also found differential activity in the PFC between the BREATH and CTRL groups. A wide area of ventral PFC was significantly activated in the CTRL group, whereas only a limited area of dorsolateral PFC was significantly activated in the BREATH group. These results suggest that focused attention to breathing increases the perseverance and concentration in difficult situations, and these behavioral changes may be related to the differential activation of PFC. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-016 霊長類腹側線条体–腹側淡蒼球の経路選択的抑制は接近回避葛藤タスク時の意欲を向上させる Silencing of the projection from the ventral striatum to the ventral pallidum increases motivational states under approach-avoidance conflict in nonhuman primates *OH Jungmin(1,2)、雨森 智子(2)、井上 謙一(1)、木村 慧(1)、高田 昌彦(1)、雨森 賢一(2) 1. 京都大学霊長類研究所　統合脳システム分野、2. 京都大学　ヒト生物学高等研究拠点 *Jungmin OH(1,2), Satoko Amemori(2), Ken-ichi Inoue(1), Kei Kimura(1), Masahiko Takada(1), ken-ichi Amemori(2) 1. Sys Neurosci Sect, Primate Res Inst, Kyoto Univ, 2. Institute for the Advanced Study of Human Biology (ASHBi), Kyoto Univ Keyword: Primate, Ventral striatum, Ventral pallidum, Motivation The ventral striatum (vStr) and ventral pallidum (VP) are suggested to regulate motivation. Although it is known that motivation under potential aversive conditions is different from that with reward, the neural system responsible for each type of motivation has not yet been fully elucidated. Because the vStr has a massive projection to the VP, it is possible that the vStr-VP pathway could play a causal role in controlling a specific type of motivation. To address this issue, we trained a macaque monkey to perform Approach-Avoidance (Ap-Av) conflict and Approach-Approach (Ap-Ap) tasks. In the Ap-Ap task, the monkey chose between two rewarding options. The Ap-Av conflict task required the monkey to choose whether to accept or reject offers composed of reward and aversive air-puff to the face. Using two comparable tasks, which require different types of motivation, we investigated how potential aversion might influence the monkey’s motivation. To manipulate the vStr activity while the monkey performed the tasks, we utilized the DREADDs technique by injecting a viral vector carrying the gene of inhibitory hM4Di receptor (AAV2.1-CaMKII-hM4Di) bilaterally into the vStr. Then we infused DCZ (deschloro-clozapine) to the VP regions for selective axonal silencing of the VS neurons projecting to the VP bilaterally. The selective suppression of the vSTr-VP pathway revealed a remarkable influence on the Ap-Av conflict task. First, we found a significant increase in the motivational state to complete the Ap-Av conflict task. The manipulation also reduced the frequency of omission errors (Kolmogorov-Smirnov test, P<0.001) and the reaction time of the target selection (two sampled t-test, P<0.001), unlike the control sessions. Second, we observed an evoked pupil dilation (t-test, P<0.001), suggesting an elevation in arousal level. The suppression caused changes in the Ap-Av decision-making, such as the increased Ap choice for the rewarding offers (t-test, P<0.05) and the increased Av choice for the aversive offers (t-test, P<0.01). It could suggest reduced fluctuations of the Ap-Av decision-making. Importantly, in the Ap-Ap task, silencing the vSTr-VP pathway did not increase the motivation and arousal level. Such behavioral changes due to selective manipulation support the hypothesis that the vStr-VP pathway regulates motivation, especially in a situation with the potential aversive outcome. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-017 心拍数を皮質刺激として提示するバイオフィードバックにより、ラットは自らの心拍数をコントロールできるようになる Heartrate-feedback training enables rats to control their heartrates *吉本 愛梨(1)、松本 信圭(1,2)、池谷 裕二(1,2) 1. 東京大学大学院薬学系研究科、2. Beyond AI研究推進機構 *Airi Yoshimoto(1), Nobuyoshi Matsumoto(1,2), Yuji Ikegaya(1,2) 1. Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan, 2. Institute for AI and Beyond, The University of Tokyo, Tokyo, Japan Keyword: biofeedback, ECGs, MFB, respiration Biofeedback is a method that trains human subjects to recognize their unconscious bodily functions (e.g., heart rate, blood pressure and even brain waves) and enables the subjects to voluntarily control the functions. During the biofeedback training, sensors are attached to subjects’ bodies to detect changes in bodily parameters, and these physiological changes are transformed to a visual, auditory or haptic signal that informs subjects of the changes. The biofeedback training is now proven valid for clinical application such as cardiorespiratory therapy for the asthma and irritable bowel syndrome; however, the neural mechanism allowing for the biofeedback has not been fully clarified. Furthermore, the experimental system of biofeedback using non-verbal animals has not been well established. Herein, as a pioneer of animals’ biofeedback experiments, we established heartrate (HR)-feedback system using electrocardiograms (ECGs) of rats and provided appropriate feedback by comparing the current HR with the target HR. We extracted instantaneous HR information from ECGs and immediately presented the information to rats by electrical stimulation of the neocortex. When the current HR was higher or lower than the target HR, electrical stimulation was applied to the right or left barrel cortex, respectively. The difference between the current and target HRs was represented by the stimulation pulse number. When rats were able to maintain their HRs below the target HR for 10 s, the medial forebrain bundle, a neural pathway involved in the integration of reward and pleasure, was stimulated as a neural reward. We demonstrated that rats were able to regulate their HRs in response to the cardiac information (i.e., instantaneous HR per se). The normal resting HR of rats was about 400-500 bpm, but during feedback training, rats were able to lower the HR to about 200-300 bpm. Because HR is related to respiration and anxiety, we used this biofeedback animal model to examine the correlation with respiration, which was estimated by electrocorticograms in olfactory bulbs. We observed respiratory rhythms were changed especially while rats kept their HRs lower, suggesting that rats learned to regulate respiratory rate to control their HRs. Moreover, elevated plus maze tests were used to assess anxiety levels of the rats. On average, rats’ HRs were high in open arms before ECG-feedback training; however, the HRs became lower even in open arms after sufficient feedback training. Our data reveal the possibility that the feedback training enabled rats to decrease their HRs even in higher levels of fear and tense. Moreover, we will analyze local field potentials in the anterior cingulate cortex, whose activity is related to anxiety level, to examine neural correlates of HR-feedback-induced reduction in anxiety-like behavior. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-018 授乳期の母親を介した「だし摂取経験」が子の油に対する食欲を変化させる The exposure to dashi through the lactating mother alters appetite of offsprings for oil during their adulthood. *伏見 駿亮(1)、佐々木 努(1) 1. 京都大学大学院農学研究科食品生物科学専攻栄養化学分野 *Shunsuke Fushimi(1), Tsutomu Sasaki(1) 1. Laboratory of Nutrition Chemistry, Division of Food Science and Biotechnology Graduate School of Agriculture, Kyoto University Keyword: appetite, bonito broth [Background] Japanese diets are eaten over generations despite their low-fat content. Although the ingestion of dashi, a characteristic of the Japanese diet, has been reported to have a negative correlation with indicators of obesity, how dashi controls remains elusive. Preliminary results showed that the experience of consuming bonito broth after weaning had no effect on appetite. Therefore, in order to examine the possibility that there is an important period prior to weaning, this study hypothesized that maternal exposure to bonito broth affects children's appetite. [Methods 1] Pregnant mice (C57BL6) were exposed to 10% concentrated bonito broth for a specific period (embryonic & lactation, embryonic, lactation). The control (non-exposed) group received water ad libitum. After pups reached adulthood, a licking test was performed to assess appetite in terms of palatability and motivation. To assess the appetite for oil., a 2.5% solution of corn oil was presented in the light phase for 15 minutes to assess the appetite for oil. [Results 1] Burst size and the number of bursts, representing palatability and motivation, respectively, were significantly increased in the embryonic & lactation and the lactation groups compared to the non-exposed and the embryonic groups. [Methods 2] Glutamic acid contained in dashi has been reported to have a suppressive effect on food intake. In order to evaluate whether results 1 is due to the ingredients of dashi or simply to glutamate, we evaluated the amount of food consumed by mothers after giving birth. The food used was normal chow CE-2, which was placed on the lid of the cage and changed every other day. [Results 2] There was no significant difference in maternal food intake between the inexperienced and lactating groups. [Conclusion] The exposure to dashi through the lactating mother alters appetite of offsprings for oil during their adulthood.　And dietary intake was not involved in this mechanism. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-019 運動パフォーマンスの長期的変動に関わる神経基盤 Neural substrates of the long-term fluctuation in motor performances: a longitudinal fMRI study in a single human *菅原 翔(1)、臼田 升(1)、福山 博幸(2)、雨宮 清美(2)、西村 幸男(1) 1. 東京都医学総合研究所、2. 東京都立松沢病院 *Sho K Sugawara(1), Noboru Usuda(1), Hiroyuki Fukuyama(2), Kiyomi Amemiya(2), Yukio Nishimura(1) 1. Tokyo Metropolitan Institute of Medical Science, 2. Tokyo Metropolitan Matsuzawa Hospital Keyword: Long-term fluctuation, Motor performance, functional MRI The performance of even well-trained motor skills varies across time, even from day to day. The performance fluctuation is decomposed into short-term (i.e., within-session) and long-term (i.e., between-session) fluctuations. We previously found that premovement activity in the ventral midbrain (VM) is associated with the within-session fluctuation in peak force (PF). However, due to the difficulty of repetitive neuroimaging in the same person for months, the neural substrates of long-term performance fluctuation are largely unknown. Here, to address this issue, we repeatedly measured the task-related activity in a single human across eight months using fMRI. One male subject participated in thirty fMRI sessions for eight months. In each fMRI session, he underwent a ready-set-go task which asked him to prepare to respond at the ready cue and to grip the force device as quickly as possible at the go cue. To investigate the neural substrates of the long-term performance fluctuation, six performances were defined: mean reaction time (RT) and PF, variability in RT and PF, omission rate, and false-start rate. To avoid the multicollinearity among these performances, principal component analysis was conducted. Two principal components explained more than 80 % of the variance. The first component (PC1) was related to longer mean RT, highly variable RT, and many omissions, reflecting less task engagement. The second component (PC2) was mainly contributed from greater mean PF and less false-start, reflecting the controllability. Premovement activities in task-irrelevant regions including the left ventral premotor area (PMv) and bilateral middle temporal gyrus varied with PC1 score reflecting less task engagement. Contrary, higher engagement (i.e., less PC1 score) was related to higher premovement activities in task-relevant cortical regions including the contralateral primary motor cortex, bilateral supplementary motor area, bilateral dorsal premotor area (PMd) as well as right putamen and bilateral VM. Although we did not find the brain region related to higher controllability (i.e., higher PC2 score), premovement activities in the contralateral sensorimotor cortex, bilateral PMd, and right cerebellum cortex. These results suggest that pre-movement activities in the task-relevant motor cortex are necessary for appropriate task engagement, but too much activation could disrupt motor control. Repetitive fMRI for a single human shed light on the neural mechanisms of the long-term performance fluctuation. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-020 目標指向的に行動するための意欲の制御におけるVTAドーパミンニューロンの多面的な役割 Multifaceted role of ventral tegmental area dopamine neurons in motivation of goal-directed behavior: An ionotropic chemogenetic study *井口 善生(1)、加藤 成樹(1)、小林 和人(1) 1. 福島県立医科大学 *Yoshio Iguchi(1), Shigeki Kato(1), Kazuto Kobayashi(1) 1. Fukushima Medical University Keyword: dopaminergic system, motivation, behavioral economics, ionotropic chemogenetics Neuromodulation of various brain regions by dopamine (DA) originated from the ventral tegmental area (VTA) is thought to play a critical role in learning and motivation in humans and animals, and thus has been the subject of much research to date. In particular, understanding the detailed role of the dopamine system in regulating the motivation for goal-directed behavior acquired through learning will lead to the elucidation of the pathogenesis of anhedonia and amotivation shared by many psychiatric disorders and the development of the treatments for them. To achieve an experimental system of high translational value, we have attempted to combine the ligand-dependent manipulations of the VTA DA neuron activity based on genetic modification techniques with assessing the effects of this intervention on the motivation-related parameters estimated using a behavioral economics demand-supply model. Our original Drd2-Cre rat line induces Cre expression not only in the indirect pathway projection neurons in the striatum, but also in authentic VTA DA neurons that express dopamine D2R as autoreceptors. We injected AAV vectors that induce Cre-dependent expression of C. elegans chloride channels (GluCl) into the Drd2-Cre rats' VTA and reversibly inhibited the activity of VTA DA neurons expressing GluCl in a ligand (ivermectin)-dependent manner. Inhibition of the neuronal activity increased the demand elasticity in food-reinforced lever pressing when the price of the reward (the number of lever presses required to obtain a certain amount of food reward) was systematically increased within a session, and also increased reward intake when the price was theoretically zero, revealing the multifaceted role of VTA DA neurons in the motivational control of goal-directed behavior. To understand the detailed circuit mechanism underlying the behavioral function, we examined the functional projection of D2R-positive VTA DA neurons to the striatum. We injected AAV vectors that induce Cre-dependent expression of Drosophila melanogaster cation channels (IR84a/IR8a complex) into the VTA of Drd2-Cre rats, and reversibly excited the activity of VTA DA neurons expressing IR84a/IR8a in a ligand (phenylacetic acid)-dependent manner. Then, the changes in the extracellular dopamine level in the nucleus accumbens (NAc) shell/core and dorsomedial striatum (DMS) were examined by a microdialysis method. Since the results of the experiment suggested that the NAc core and DMS are candidates for the functional terminals of the D2R+ VTA DA neurons, we will examine the effects of selective inhibition of these circuits by the GluCl system on the motivational parameters. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-021 イソフラボンによる閉経モデルマウスの高脂肪食に対する嗜好性の抑制効果 Preventive effect of isoflavone on developing the tendency towards high-fat diet in ovariectomized mice: a menopausal model for women *小宇田 智子(1)、牧野 千恵(1)、手塚 雄太(1)、高橋 司(2)、高成 準(2)、明石 眞言(1) 1. 東京医療保健大学、2. 株式会社アミノアップ *Tomoko Koda(1), Chie Makino(1), Yuta Tezuka(1), Tsukasa Takahashi(2), Jun TYakanari(2), Makoto Akashi(1) 1. Tokyo Healthcare University, 2. Amino Up Co., Ltd Keyword: conditioned place preference , high-fat diet, menopausal women, isoflavones The hyper-palatable foods can activate our brain reward neurocircuitry and create a highly rewarding experience; these foods enable us to stop eating even when we feel full. Previously, we showed that the ovariectomized mice eating high-fat diet are prone to high-fat diet using behavior test. Genistein is the major isoflavone in soybeans and its estrogenic activities have been reported in many articles. Since isoflavones in soybeans are generally present in their glucoside form, these cannot be absorbed in our body because of binding sugar. In this study, we aimed to examine the effects of a novel aglycone isoflavone-rich extract, genistein combined polysaccharide (GCP) on high-fat diet tendency in experimental menopausal women model mice. GCP was provided from Amino Up Company, Sapporo, Japan. Ovariectomized (OVX) female C57BL/6J mice were divided randomly into 4 groups: low-fat diet group (control), high-fat diet group, high-fat diet + 2 % GCP group (2 % group), and high-fat diet + 4 % GCP group (4 % group). These mice were bred for 139 days. At days 132, 134 and 136, mice were confined to light chamber with high-fat diet for 20 min. At days 133, 135 and 137, mice were confined to dark chamber with low-fat diet for 20 min. At day 138, conditioned place preference (CPP) was evaluated the difference in the time spent, moving distance and rearing times between the light and dark chambers. At day 139, the brain tissues were collected under anesthesia for histological assay to assess the expressions of dopamine receptor d1 (D1) and dopamine transporter (DAT) in ventral tegmental (VTA) and nucleus accumbens (ACB) area. In the CPP test, the high-fat diet and the 2 % groups stayed in the light zone more than the control group. In the 4 % group, the time spent in the light zone was reduced compared to the high-fat diet group. In addition, the 4 % group moved longer distance in light zone than high-fat group. Our results suggest that there may be a tendency towards an increased fat-diet in menopausal woman and GCP may prevent the tendency for high-fat diet. To study the effects on neurologic function, histological and molecular-biological studies are now in progress. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-022 ビタミンB1誘導体は青斑核-皮質カテコラミン系を介して身体活動を促進する Vitamin B1 derivative enhance physical activity via coeruleo-cortical catecholaminergic pathways *秦 俊陽(1,2)、平賀 大一(1,2)、グレニエ フランソワ(1)、岡本 正洋(1,3)、松井 崇(1,3)、征矢 英昭(1,3) 1. 筑波大学運動生化学研究室、2. 日本学術振興会、3. 筑波大学体育系ヒューマン・ハイ・パフォーマンス先端研究センター *Toshiaki Hata(1,2), Taichi Hiraga(1,2), François Grenier(1), Masahiro Okamoto(1,3), Takashi Matsui(1,3), Hideaki Soya(1,3) 1. Exercise Biochem & Neuroendocrinol, Univ of Tsukuba, Ibaraki, Japan., 2. Japan Society for the Promotion of Science, Tokyo, Japan., 3. Sport Neuroscience Division, Advanced Research Initiative for Human High Performance (ARIHHP), Univ of Tsukuba, Ibaraki, Japan. Keyword: Vitamin B1 derivative, physical activity, coeruleo-cortical pathways Physical activity is beneficial for maintaining healthy life, yet current pandemic makes it impossible to maintain such the active lifestyle. There are various brain regions involved in physical activity, for example the limbic system, frontal cortex, and motor cortex, etc. Thiamine tetrahydrofurfuryl disulfide (TTFD), a vitamin B1 derivative, is known to promote voluntary physical activity by increasing dopamine release in the medial prefrontal cortex (mPFC) (Saiki et al., 2018) but other regions have not been fully investigated. Here, we aimed to clarify brain regions where TTFD activate and the monoaminergic connections to these regions. Male Wistar rats were acclimated for a week in a cage, where animals are then allowed to monitor voluntary physical activity with either i.p. injection of TTFD (50mg/kg) or saline. The brains were removed 100 minutes later, and brain sections were prepared for immunohistochemical analysis. The neural activity was evaluated through the number of double positive cells of c-Fos/NeuN. We found the TTFD activate the motor cortex (MC), anterior cingulate cortex (ACC), medial prefrontal cortex (mPFC). Furthermore, the activity of monoaminergic neurons was confirmed by the number of double positive cells of c-Fos/TH or c-Fos/5-HT. As a result, TTFD activated ventral tegmental area (VTA), locus coeruleus (LC), dorsal raphe nucleus (DRN). Finally, TTFD increases the number of c-Fos expression of neurons projecting from the LC to the mPFC labeled with retrograde tracers. These results lead us suggest that the coeruleo-cortical catecholaminergic pathways is predominantly involved in TTFD-enhanced voluntary physical activity. In the next step, we will need to solid evidence by specific inhibition of this pathway (e.g., by DREADD or optogenetics). 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-023 5-HTTLPRのs/s遺伝子型を持つ薬物未使用初発うつ病患者における扁桃体サブ領域体積と血中物質の関係: 探索的研究 Relationship between Volume of Amygdala Subregions and Serum levels of catecholamine metabolites, BDNF, or cortisol in Patients with First-Episode, Drug-Naive Major Depression in Japanese with the s/s genotype of 5-HTTLPR: an Exploratory Research *岡本 直通(1)、渡邊 啓太(2)、池ノ内 篤子(1)、井形 亮平(1)、小西 勇輝(1)、手銭 宏文(1)、夏山 知也(1)、藤井 倫太郎(3)、岸 太郎(3)、岩田 仲生(3)、吉村 玲児(1) 1. 産業医科大学精神医学教室、2. 京都大学オープンイノベーション機構、3. 藤田医科大学　精神神経科学講座 *Naomichi Okamoto(1), Keita Watanabe(2), Atsuko Ikenouchi(1), Ryohei Igata(1), Yuki Konishi(1), Hirofumi Tesen(1), Tomoya Natsuyama(1), Rintaro Fujii (3), Taro Kishi(3), Nakao Iwata(3), Reiji Yoshimura(1) 1. Department of Psychiatry, University of Occupational and Environmental Health, Japan, 2. Kyoto University Open Innovation Institute, 3. Department of Psychiatry, Fujita Health University. Keyword: amygdala, major depression, cortisol, 5-HTTLPR Individuals with the s/s genotype of the serotonin transporter gene-linked promoter region (5-HTTLPR), which occurs with high frequency in the Japanese population, show diagnosable depression in association with stressful life events compared with those with the s/l or l/l genotypes. The amygdala is active when we feel emotions such as anxiety or fear, however in patients with major depression, the activity and volume of the amygdala is known to be different to that of healthy people. We prospectively investigated differences in total amygdala area and subvolume in 25 Japanese patients with s/s genotype and drug-naïve first-episode depression and 46 healthy subjects. We also examined correlations between serum levels of HVA, MHPG, BDNF, and Cortisol and amygdala total volume and subvolume in patients with depression and healthy control. Whole-brain volume and amygdala subvolume tended to be lower in patients with depression than in healthy subjects, but the difference was not statistically significant. There was a negative correlation between serum HVA and left central nucleus and left paralaminar nucleus volumes (β=-0.387, p=0.042) (β=-0.255, p=0.049). Serum cortisol was a positive correlation between the left medial nucleus (β=0.548, p=0.0082) and right central nucleus volume (β=0.347, p=0.043). There was a significant difference effect on the left medial nucleus and serum levels of cortisol in HC and MD groups (p = 0.013 for interaction). In Japanese patients with depression with the s/s genotype of 5-HTTLPR, amygdala subregional volume and serum levels of catecholamine metabolites and cortisol could be associated. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-024 急性ストレス経験後の安静時機能結合にレジリエンスの個人差が表現されている Individual differences in psychological resilience are revealed by the dynamics of the resting-state functional network driven by an acute stress experience *渡邊 言也(1)、Ruedeerat Keerativittayayut(2)、中原 潔(1)、竹田 真己(1) 1. 高知工科大学、2. HRHプリンセスチュラブホーン医科⼤学 *Noriya Watanabe(1), Ruedeerat Keerativittayayut(2), Kiyoshi Nakahara(1), Masaki Takeda(1) 1. Kochi University of Technology, 2. HRH Princess Chulabhorn College of Medical Science, Chulabhorn Royal Academy Keyword: FMRI, STRESS, RESILIENCE, TEMPORAL DYNAMICS Psychological resilience refers to a person’s ability to adapt successfully to acute stress, trauma, or more chronic forms of adversity (Feder et al., 2009). This ability is important to keep a mentally healthy life, and lower resilience increases the risk of mental diseases (Southwick & Charney, 2012). Although there is a non-negligible individual difference in resilience, its neurophysiological mechanism has not been clarified, especially in human brains. The current study evaluated the individual differences in resilience by resting-state functional magnetic resonance imaging (rsfMRI) with one acute stress experience. In the experiment, subjects (n = 102, male : female = 77 : 25, Age = 20.10 ± 1.65 years) engaged in five successive sessions within 2 hours. Individual resilience level was evaluated by the Connor-Davidson resilience scale (CD-RISC: Connor and Davidson, 2003). After collecting the baseline session, all subjects experienced the cold-pressor-test (CPT: Schwabe et al., 2008) as the acute stress. Then, additional four sessions were run intermittently within 90 min. ROI-to-ROI functional connectivity (FC) analysis identified two functional networks correlated with the CD-RISC scores (cluster-level correction, PFDR < 0.05) at a specific time point; 60-75 min from CPT. The first network involves the bilateral insula, bilateral dorsal anterior cingulate cortex, right orbitofrontal cortex, and right dorsomedial/lateral prefrontal cortex. FC strength within this network was negatively correlated with the individual CD-RISC scores. The second network involves the right inferior frontal cortex, bilateral precuneus, and posterior cingulate cortex. FC strength within this network was positively correlated with the scores. Next, to understand the role of the subcortex system on psychological resilience, we evaluated ROI-based BOLD signal fluctuations by the fractional amplitude of low-frequency fluctuations (fALFF) analysis. Based on previous reports related to resilience, seven ROIs in subcortical regions were selected (Watanabe & Takeda, 2021). Among these ROIs, we found that the CD-RISC score was correlated with (1) fALFF in the locus coeruleus (LC) at 15-30 min from CPT when the stress-driven cortisol level was peaked, and (2) fALFF in the posterior hippocampus (HIPp) at 60-75 min from CPT when the resilience-related FC networks were activated. These collective results suggest novel resilience-dependent functional brain networks under stress in humans. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-025 扁桃体基底外側核ニューロンの膜電位と新皮質徐波との位相同期機構の解明 Phase-locking mechanism of membrane potentials of basolateral amygdalar neurons and the neocortical slow oscillations *佐藤 由宇(1)、池谷 裕二(1,2) 1. 東京大学大学院薬学系研究科、2. Beyond AI 研究推進機構 *Yu Sato(1), Yuji Ikegaya(1,2) 1. Grad Sch Pharmaceut Sci, Univ of Tokyo, Tokyo, Japan, 2. Institute for AI and Beyond, University of Tokyo, Tokyo, Japan Keyword: BASOLATERAL AMYGDALA, SLOW OSCILLATION, WHOLE-CELL RECORDING, DEPOLARIZATION The basolateral amygdala (BLA) shows synchronized activity with the neocortex in various oscillatory bands, including slow oscillations, a form of <1-Hz oscillations that occur dominantly during slow-wave sleep or under anesthesia. BLA extracellular potentials oscillate in synchrony with neocortical slow oscillations and are suggested to play a role in emotional memory formation. However, it is technically difficult to record neuronal membrane potentials in deep brain regions, such as the BLA, and it remains unclear how this synchronization arises from the dynamics of BLA neuronal membrane potentials. Since subthreshold membrane potentials determine the process from synaptic input to somatic output, elucidating the mechanism of the synchronization is important for understanding the neural circuits involved in emotional memory formation. We recorded local field potentials from the neocortex simultaneously with membrane potentials in BLA neurons using a new method that enabled whole-cell recordings from deep brain regions in vivo. We found that BLA neurons periodically depolarized along neocortical slow oscillations. The timings of the depolarizations of BLA neurons varied depending on the lengths of neocortical UP states, but the depolarizations tended to occur in later phases of individual UP states and seemed to be time-locked to the offsets of UP states. We also confirmed the time-locked synchronization using optical imaging of somatic calcium events of multiple BLA neurons. To determine the neural source of the depolarization of BLA neurons, we retrogradely labelled BLA-projecting neurons with channelrhodopsin-2 using retrograde adeno associated virus and recorded their firing activities. As a candidate region involved in BLA neuronal depolarizations locked to neocortical slow oscillations, we focused on the prelimbic cortex, whose projections to the BLA contribute to the acquisition of fear memory. These results provide insights into neuronal mechanisms by which slow oscillations are synchronized between the BLA and the neocortex and may lead to the elucidation of the mechanism of emotional memory formation. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-026 Dominant role of primary motor cortex in driving reward-related facial expressions *Wanru Li(1,2), Takashi Nakano(3), Takayuki Yamashita(1) 1. Dept Phys, Grad Sch Med, FHU, Toyoake, Japan, 2. Dept Neural Regul, Grad Sch Med, Univ of Nagoya, Nagoya, Japan, 3. Dept Comput Biol, Grad Sch Med, FHU, Toyoake, Japan Keyword: facial expressions, orofacial movements, reward, primary motor cortex Animals exhibit facial expressions correlated with various internal and emotional states. However, the neural mechanisms underlying facial expressions are poorly understood. Here, using a transgenic mouse line expressing channelrhodopsin 2 in dopamine neurons (DAT-Cre; LSL-ChR2 mice), we investigated the reward-related facial movements of mice. Blue light stimulation (5 ms x20, 20 Hz) through an optical fiber were targeted to dopamine (DA) neurons in the ventral tegmental area (VTA), and orofacial movements of the head-fixed mice were simultaneously filmed from a dorsal position using a high-speed video camera (500 frames per second). We found that optogenetic stimulation of VTA-DA neurons (oDAS), which is rewarding for the mice, often induced stereotypical motor actions in their whisker and nose. Furthermore, after learning the association between a sound cue presentation and oDAS, mice exhibited a transient whisker protraction and nose twitch immediately after the sound cue onset. Interestingly, inhibition of dopamine D1 receptor (D1R) abolished only oDAS-induced facial movements but did not affect cue-locked facial movements. However, pharmacological inhibition of the whisker primary motor cortex (wM1) largely attenuated both oDAS-induced and cue-locked whisker/nose movements. In vivo multi-unit recordings from wM1 using silicone probes further revealed that a fraction of neurons shows activities correlated with either oDAS-induced or cue-locked whisker movements. Thus, our findings suggest that D1R-dependent and -independent neuronal circuits drive reward-related facial expressions in mice, both of which depend on the activity of wM1. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-027 ミクログリアのストレス応答を担う転写・エピゲノム制御 Transcriptional and epigenetic regulations of microglia for stress responses *谷口 将之(1)、松下 和敏(1)、三島 零(1)、北岡 志保(2)、工樂 樹洋(3,4)、門田 満隆(3)、古屋敷 智之(1) 1. 神戸大学大学院医学研究科 薬理学分野、2. 兵庫医科大学 薬理学講座、3. 理化学研究所 生命機能科学研究センター 分子配列比較解析チーム、4. 国立遺伝学研究所 分子生命史研究室 *Masayuki Taniguchi(1), Kazutoshi Matsushita(1), Rei Mishima(1), Shiho Kitaoka(2), Shigehiro Kuraku(3,4), Kadota Mitsutaka(3), Tomoyuki Furuyashiki(1) 1. Div Pharmacol, Grad Sch Med, Kobe Univ, Kobe, Japan, 2. Dept Pharmacol, Hyogo Col Med, Kobe, Japan, 3. Lab Phyloinformatics, RIKEN BDR, Kobe, Japan, 4. Molecular Life History Laboratory, NIG, Mishima, Japan Keyword: Microglia, Stress, Epigenetics Microglial activation is crucial for stress pathology in mental illnesses. However, how stress alters microglia remains elusive. Using transcriptome and epigenome analyses in two stress-related brain regions, the medial prefrontal cortex and the nucleus accumbens, we found that microglia shows brain region-specific (local) and non-specific (global) transcriptional responses to repeated social defeat stress. Local microglial responses emerged in a subset of H3K27ac-marked super-enhancers after acute social defeat stress, leading to adjacent gene expressions after chronic social defeat stress. By contrast, global microglial responses emerged in another subset of the super-enhancers and adjacent gene expressions after acute and chronic social defeat stress. We speculated distinct transcription factors involved in the local and global responses and identified a putative transcription factor sensing a stimulus to induce the global response. Individual variability of stress-susceptibility was encoded in the global microglial response, including the increase in an innate immune receptor crucial for stress-induced emotional disturbances. These findings show that chronic social defeat stress induces local and global transcriptional and epigenetic responses in microglia to confer neuroinflammation with the brain region specificity and individual variability of stress-susceptibility. Thus, microglia may integrate local and global stress responses and determine its activation state and behavioral consequence. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-028 交配前に心理的ストレスを受けた父親マウスに由来する次世代仔マウスの情動行動変化はグルココルチコイド受容体拮抗薬で防ぐことができる Glucocoliticoid receptor antagonist (RU-486) prevent a defect emotional behavior observed in offspring derived from paternal mouse subjected psychological stress. *清水 紀之(1)、近久 幸子(1)、志内 哲也(1)、勢井 宏義(1) 1. 徳島大院医歯薬学統合生理 *Noriyuki Shimizu(1), Sachiko Chikahisa(1), Tetsuya Shiuchi(1), Hiroyoshi Sei(1) 1. Dept Integ Physiol, Inst of Biomed Sci, Grad Sch Univ of Tokushima, Tokushima, Japan Keyword: Paternal , Psychological stress, Offspring, Emotional behaviors Psychological stress subjected parent has the potential to influence a formation of brain functions related to regulate emotional behaviors in next generations, however, these mechanisms are still unclear. In this study, we investigated whether RU-486, a glucocolticoid receptor antagonist, would act the paternal psychological stress, and cause how the change of offspring’s emotional behaviors. Male C57/BL6 mice (aged 8-9 weeks) were subjected to restraint stress for 2 hours by using 50-mL conical centrifuge tube. This treatment was continued for 2 weeks. RU-486 was injected to mice before 30 min of restraint stress. After a final restraint stress, each male mouse was mated with same aged virgin female for only 2 days. When the offspring has reached 10 weeks of age, their emotional behaviors were evaluated by several behavioral analysis. Restraint-stress offspring mice showed a depression-like behavior, and a strong fear response to electrical stimulation compared with non-stress offspring mice. These emotional behavior changes observed restraint-stress offspring were recovered by RU-486 injection to paternal before restraint stress. On the other hands, offspring derived from paternal mouse subjected an injection of dexamethasone, a glucocorticoid receptor agonist, were showed an increased depression-like behavior. The above-mentioned results indicate a possibility that the continued psychological stress before mating would affect next generation offspring’s emotional behaviors via a glucocolticoid signaling. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-029 Brian circuits of cognitive control in emotional inference *Xiaowei Gu(1), Joshua Johansen(1) 1. CBS, RIKEN, Wako-shi, Japan Keyword: emotional learning, cognitive control, fear inference, rats To survive and thrive in the world organisms need to learn to avoid danger, sometimes based on limited experience. Learning which combination of sensory stimuli predict impending danger based on prior aversive experience facilitates the recruitment of rapid defensive responses when animals encounter similar sensory contexts again. However, humans and other species can also use inference to extrapolate the circumstances surrounding directly experienced aversive events to other related contexts and sensory patterns which weren’t a part of the original traumatic experience. Brain systems of emotion have evolved to enable associative aversive learning and the brain mechanisms of directly experienced aversive learning have been well studied. However, how brain circuits of cognition and emotion interact to implement emotional inference is not known. In this study, we developed a rodent behavioral model to study inference during aversive learning. Calcium imaging was used in the amygdala (a brain region important for the formation and storage of emotional memories) and in the medial prefrontal cortex (mPFC, critical for cognitive processing and emotion) to detect the neural representation of emotional inference during and after aversive learning. Optogenetics was used to inhibit the projection from mPFC to amygdala during different phases of fear learning to identify the causal role of information flow from mPFC to amygdala in the formation and expression of emotional inference. Our results show that both inferred and directly experienced fear memories are encoded in the neural activity of mPFC and amygdala neuronal populations. Notably, mPFC neural coding of sensory stimuli which were directly associated with the aversive experience and those which were indirectly associated through inference reorganized and became more specific following aversive learning. Moreover, the projections of mPFC to the amygdala were necessary to produce inferred memory expression, specifically during memory recall. In summary, these results are beginning to reveal the mechanisms of how cognitive and emotional brain systems interact to produce emotional inference and coordinate corresponding defensive behaviors. This study sheds light on how higher order emotional processing is organized in the brain and has important implications for our understanding of trauma related psychiatric conditions (e.g. PTSD) which are associated with disturbances in cognitive appraisals of threat leading to inappropriate emotional expression. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-030 主観的な感情評価が感情そのものへ与える影響：脳波の前頭左右非対称性による検討 Effect of subjective evaluation of emotion on the emotion itself: Investigation by Frontal Alpha Asymmetry using Electroencephalography *伊東 正登(1)、髙橋 徹(1)、栗原 勇人(1)、大須 理英子(1) 1. 早稲田大学 *Masato Ito(1), Toru Takahashi(1), Yuto Kurihara(1), Rieko Osu(1) 1. Waseda University Keyword: Emotion Recognition, EEG, Frontal Alpha Asymmetry, Self-focused Attention In studies of emotion recognition, subjective evaluation of emotion is conducted to confirm whether the targeted emotion is elicited. However, in recent years, it was shown that subjective evaluations affect the state of attention to a stimulus, resulting in differences in the event-related potential of the electroencephalogram (EEG). In addition, in psychology, it has long been stated that self-focused attention enhances emotional experiences. This suggests that subjective evaluation of emotion may alter emotional states and thus EEG. We tested this hypothesis by examining the effect of subjective evaluation of emotion on Frontal Alpha Asymmetry (FAA). We hypothesized two types of effects of subjective evaluation of emotion on EEG: (1) immediate effects after stimulus presentation when subjective evaluation is performed, and (2) persistent effects that appear during subsequent measurements. EEGs were recorded from 40 participants with 28 channels. FAA was calculated as FAA = log(F4) - log(F3) by the alpha frequency power. The experiment consisted of two conditions: in the “with evaluation” condition, participants were presented with the negative high-arousal stimulus (1 s) from The International Affective Picture Systems (Lang et al, 2008), and then made subjective evaluations of their emotional states by pressing buttons 1 to 9 according to the Self-Assessment Manikin (Bradley et al, 1994). In the “without evaluation” condition, participants simply pressed a button on a number displayed on the screen. This single trial was conducted 25 times in each condition. To examine the immediate effects, the experimental group performed the "with evaluation" condition, and the control group performed the "without evaluation" condition. Subsequently, the "without evaluation" condition was conducted for both groups to examine the persistent effects. By comparing the FAA in each condition between the two groups, the immediate and persistent effects of subjective evaluation were examined. Welch's t-test confirmed a significant decrease in FAA of the immediate effects. This means that negative emotions were activated by subjective evaluations of emotion. No significant effect was found for the persistent effects. The results suggest that subjective evaluation of emotion may have an immediate activating effect on emotion. This study may provide caution in emotion estimation studies and may also provide neuroscientific support for the effects of self-focused attention. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-031 ノルアドレナリン作動性神経選択的AAVベクターの構築と社会性ストレスにより活動する細胞の軸索分布解析 Development of a noradrenergic neuron-selective AAV vector and brain-wide axonal mapping of social stress-responsive noradrenergic neurons *勢力 薫(1)、前田 駿介(1)、藤本 柚香(1)、山田 翔平(1)、平戸 祐充(1)、笠井 淳司(1)、橋本 均(1,2,3,4,5) 1. 大阪大学大学院薬学研究科、2. 大阪大学大学院連合小児発達学研究科、3. 大阪大学データビリティフロンティア機構、4. 大阪大学先導的学際研究機構、5. 大阪大学大学院医学系研究科 *Kaoru Seiriki(1), Shunsuke Maeda(1), Yuzuka Fujimoto(1), Shohei Yamada(1), Yumi Hirato(1), Atsushi Kasai(1), Hitoshi Hashimoto(1,2,3,4,5) 1. Grad Sch Pharmaceutical Sci, Osaka Univ, Osaka, Japan, 2. United Grad Sch Child Dev. Osaka Univ, Osaka, Japan, 3. Inst Datability Sci, Osaka Univ, Osaka, Japan, 4. Inst for Open Transdisciplinary Res Initiatives, Osaka Univ, Osaka, Japan, 5. Grad Sch of Med, Osaka Univ, Osaka, Japan Keyword: locus coeruleus, noradrenaline, imaging, AAV The locus coeruleus (LC) noradrenergic (NA) system is considered to play important roles in emotion and stress response. A variety of recent studies show that functionally distinct LC-NA neurons send segregated efferent projections in a target brain region-specific manner. Although stress-related noradrenergic regulation in the amygdala is well characterized, contribution of other brain regions is less unclear. In the present study, we investigated the output brain regions of stress-responsive LC-NA neurons throughout the brain by activation-dependent axonal labeling and whole-brain imaging. For this purpose, we first developed a novel adeno-associated viral (AAV) vector for NA neuron-selective gene expression. By stereotaxic injection of the AAVs into the LC, we observed that 92% of cells labeled with fluorescent proteins were positive for tyrosine hydroxylase. The fluorescent intensity of labeled axons by these AAVs with axon-targeting fluorescent proteins were sufficient for microscopic detection without immunohistochemical signal amplification. Using these AAVs with Cre recombinase-dependent gene expression system and TRAP2 (c-Fos-2A-CreERT2) mice, we performed activation-dependent and LC-NA-selective labeling followed by whole-brain fluorescent imaging. We found that the social defeat stress-responsive LC-NA neurons innervated not only amygdala-related nuclei, but also other brain regions, e.g. ventral hippocampus and piriform cortex. Future study investigating functional roles of NA release in these brain regions will contribute to understanding of complex organization of LC-NA systems in stress response, and the LC-NA-selective AAV can be an alternative for LC-NA-selective labeling method such as Cre-driver mouse lines. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-032 嫌悪刺激に対する左眼窩前頭皮質の活動と低レジリエンスおよび抑うつ傾向の関連性 Association of low resilience and depressive tendency with activity in the left orbitofrontal cortex to aversive stimuli *笹岡 貴史(1)、小野 健太郎(1)、山脇 成人(1) 1. 広島大学　脳・こころ・感性科学研究センター *Takafumi Sasaoka(1), Kentaro Ono(1), Shigeto Yamawaki(1) 1. Center for Brain, Mind, and KANSEI Sciences Research, Hiroshima University Keyword: RESILIENCE, ORBITOFRONTAL CORTEX, DEPRESSION, FMRI Resilience refers to the ability to recover from stress and has received increasing attention due to its association with mental health. Although previous studies have examined the neural basis of individual differences in resilience (e.g., Waugh et al., 2008), the relationship among the neural basis of resilience traits, individual differences in depressive tendencies, and interoception has not been clarified. In the present study, we investigated the neural basis of individual differences in resilience, mental health, and interoception. Using functional magnetic resonance imaging to measure brain activity during the presentation of emotion-eliciting images, we examined the correlation between brain activity and scores obtained from questionnaires on resilience, depression, and interoception. During the experiment, participants were presented with pleasant, unpleasant, and neutral images for 3 seconds, after which they rated their emotional valence and arousal elicited by the image using an affect grid (Russell et al., 1989). In addition, responses from each participant to questionnaires on resilience (short version of the Resilience Scale, RS14; Nishi et al., 2010; Bidimensional Resilience Scale, BRS; Hirano, 2010), depressive tendency (Beck Depression Inventory, BDI; Beck, et al., 1996), and interoceptive awareness (Multidimensional Assessment of Interoceptive Awareness, MAIA; Mehling et al., 2012) were reviewed. We then examined the brain regions showing the activity correlated with scores attained from the questionnaires. In the unpleasant versus pleasant images brain activity contrast, we found that the activations in the left orbitofrontal cortex, left middle insular cortex, anterior pregenual cingulate gyrus, and left superior occipital gyrus were negatively correlated with the resilience scores derived by RS-14. However, no brain region activation showed a correlation with the resilience scores derived by the BRS. In contrast, the activity in the left orbitofrontal cortex in the unpleasant versus pleasant images brain activity contrast showed a positive correlation with depressive tendency as measured by the BDI. Given that the orbitofrontal cortex is involved in inhibition of pain (Wager et al., 2004) as well as emotion regulation (Lévesque et al., 2003), our results suggest that the depressive and low resilient individuals recruit the orbitofrontal cortex for emotional inhibition because of their high reactivity to the aversive stimuli. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-033 内側前頭前野前辺縁皮質－扁桃体基底外側核神経回路は不安様行動の発現に関与する Specific activation of excitatory prelimbic cortico-amygdala pathway modulates innate fear, but not conditioned fear in mice. *河南 絢子(1)、西田 萌乃(1)、山田 大輔(1)、飯尾 啓太(2)、長瀬 博(2)、斎藤 顕宜(1) 1. 東京理科大学　薬学部　薬理学研究室、2. 筑波大学　国際統合睡眠医科学研究機構 *Ayako Kawaminami(1), Moeno Nishida(1), Daisuke Yamada(1), Keita Iio(2), Hiroshi Nagase(2), Akiyoshi Saitoh(1) 1. Lab Pharmacol, Fac Pharm Sci, Tokyo Univ of Science, 2. IIIS, Univ of Tsukuba Keyword: anxiety , neural circuit, basolateral nucleus of the amygdala, prelimbic medial prefrontal cortex [Introduction] Prelimbic subregion of the medial prefrontal cortex (PL) has been shown to contribute to express fear response and/or anxiety-like behavior in rodents. In addition, changes in the PL activity have also been demonstrated in patients suffer from major depression and anxiety disorders. We previously reported that the artificial activation of the PL by local perfusion of a sodium channel activator, veratrine, induced anxiety-like behavior in the mouse open-field test, and that similar activation of the PL significantly upregulated the expression of c-Fos immunoreactivity in the amygdala. These results suggested that excitatory projections from PL to amygdala play a role in the regulation of anxiety-like behavior. Furthermore, we reported that the veratrine-induced anxiogenic effect was attenuated by intra-PL perfusion of a selective delta-opioid receptor (DOP) agonist, KNT-127. In the present study, we aimed to establish experimental conditions to examine the neurocircuitry mechanism of anxiolytic-like effect of KNT-127 in mice. [Materials & Methods] Male C57BL/6J mice (4 weeks old) were bilaterally administered with adeno-associated-virus (AAV)2-CaMKIIa-hChR2(H134R)-EYFP into the PL to specific activation of the PL-BLA projection. Five weeks later, robust expression of ChR2-EYFP was observed in the PL and basolateral nucleus of the amygdala (BLA). Then, optic fiber canula connected to wireless photo-stimulator was implanted into the BLA. In these animals, we examined innate fear (i.e. anxiety) in the open-field test and elevated-plus maze test and conditioned fear in the contextual fear conditioning test. During these tests, PL-BLA pathway were photo-stimulated at frequency of 10 Hz. [Results & Discussion] In the open field test, the percentage of time spent in the center area of the ChR2 group was significantly reduced compared to the control group. In addition, in the elevated-plus maze test, the percentage of time spent in the open arm of the ChR2 group was significantly lower than that of the control group. However, in the fear conditioning test, there was no significant difference in the freezing rate between the control and ChR2 groups. These results suggest that the projection from the PL to BLA is involved in the expression of innate fear, but not the contextually conditioned fear responses. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-034 失感情症と脳活動の関係についての研究 Research on the relationship between alexithymia and brain activity *福崎 采加(1)、平林 直樹(2)、権藤 元治(3)、伊津野 巧(2,4)、吉原 一文(2)、元村 祐貴(5) 1. 九州大学大学院統合新領域学府 ユーザー感性学専攻、2. 九州大学大学院医学研究院 心身医学、3. 九州大学病院 心療内科、4. 自然科学研究機構 生理学研究所 システム脳科学研究領域、5. 九州大学大学院芸術工学研究院 デザイン人間科学部門 *Ayaka Fukuzaki(1), Naoki Hirabayashi(2), Motoharu Gondo(3), Satoshi Izuno(2,4), Kazufumi Yoshihara(2), Yuki Motomura(5) 1. Department of Kansei Science, Graduate School of Integrated Frontier Sciences, Kyushu University, 2. Department of Psychosomatic Medicine, Graduate School of Medical Sciences, Kyushu University, 3. Department of Psychosomatic Medicine, Kyushu University Hospital, 4. Division of Cerebral Integration, Department of System Neuroscience, National Institute for Physiological Sciences (NIPS), 5. Department of Human Science, Faculty of Design, Kyushu University Keyword: Alexithymia Alexithymia is a personality trait in which a person has feelings of joy, anger, sadness and anger, but is not good at recognising or expressing them. It is known to affect health care and interpersonal relationships because it is difficult to perceive stress and the emotions of others. There has been a great deal of research into the relationship between emotional processing disorders and the brain, with a number of studies focusing on functional aspects of the brain. These studies have generally been task- or stimulus-based, and few have focused on brain function at rest. Recently, it has been shown that there are changes in brain fMRI signals even in the resting state, and that these changes are closely related to the signal processing infrastructure of the brain. Therefore, it is necessary to investigate brain functions during resting state in order to clarify the brain mechanisms of psychological factors such as alexithymia. In this study, we analyzed MRI data obtained during the resting state to investigate the relationship between alexithymia. and brain function and structure in more detail. In this study, we focused on multiple resting-state networks and investigated functional connectivity within and between networks. In addition, the results of a questionnaire (TAS20) were used to examine the relationship between alexithymia. and brain function. The results showed that functional connectivity within the default mode network, one of the resting state networks, was associated with alexithymia. Specifically, functional connectivity between the left posterior cingulate cortex and the left gaze frontal area was weaker with higher tendency to aphasia. In addition, functional connectivity between the left posterior cingulate cortex and the right inferior parietal cortex was stronger with higher tendency toward alexithymia. The present results suggest that functional connectivity differs according to the degree of alexithymia. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-035 情動に対する音楽の影響を動物モデルで調べるための実験系 An experimental system to investigate the effects of music on emotion in animal models *山木 崚太郎(1)、白松-磯口 知世(1)、石田 直輝(1)、高橋 宏知(1) 1. 東京大学大学院情報理工学系研究科 *Ryotaro Yamaki(1), Tomoyo I Shiramatsu(1), Naoki Ishida(1), Hirokazu Takahashi(1) 1. Grad Sch of Information Science and Technology, Univ of Tokyo, Tokyo, Japan Keyword: MUSIC, DOPAMINE, HEARTBEAT-EVOKED POTENTIAL Accumulating evidence suggests that the dopaminergic system is deeply involved in the music-evoked pleasure. As the next step, simultaneous recordings of dopaminergic level and emotion level at higher temporal resolution should provide a better understanding of their relationship. Herein, the present study attempted to establish an animal experiment system that consist of a measurement of rapid dopaminergic dynamics, i.e., fast-scan cyclic voltammetry (FSCV), and a measurement of a candidate for biological index of emotion, i.e., heart-beat evoked potential (HEP). Rats were implanted with a custom-made head-fix frame, which was used to fix them to the setup. For FSCV, we also implanted a carbon-fiber microelectrode into the right striatum. To obtain HEP, some screws with wires were anchored to the skull. The screw anchored at the interparietal bone was used as the ground electrode, and the screw in contact with the dura over the cerebellum was used as the reference electrode. Other screws in the frontal and parietal bones, in contact with the dura to record electrocorticogram epidurally. At the recording, sheet electrode was placed onto the chest of the animal to obtain electrocardiogram. First, we applied electrical stimulations to the right ventral tegmental area and demonstrated that FSCV signal reflected electrochemical fluctuation, which is expected to be a dopaminergic signal, with a temporal resolution at 10 Hz. Second, we succeeded to obtain putative HEP as fluctuations in grand-averaged epidural electrocorticogram in response to the heartbeats (R-peak). Finally, electrocardiogram of awake animals exhibited that heart rate tended to change when a piece of classical music was presented. Taken together, each setup will contribute to simultaneous recording of dopaminergic level and emotional change in response to music from an animal model. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-036 内受容感覚訓練効果の神経基盤の解明 Neural underpinnings of an effect of interoceptive training *関口 敦(1)、菅原 彩子(1)、勝沼 るり(1)、寺澤 悠理(2) 1. 国立精神‣神経医療研究センター　精神保健研究所、2. 慶応大学文学部心理学科 *Atsushi Sekiguchi(1), Ayako Sugawara(1), Ruri Katsunuma(1), Yuri Terasawa(2) 1. National Institute of Mental Health, National Center of Neurology and Psychiatry, 2. Psychology, Keio University Keyword: Interoception, Anterior insula Interoception is perception of afferent information that arises from anywhere and everywhere within the body (Cameron 2001). Individual differences in interoceptive awareness were associated with not only affective processing, but also decision making processing (Critchley 2013). Recently, interoceptive awareness could be enhanced by biofeedback technique (Sugawara 2020, Quadt, 2021). Given that anterior insula (AI) cortex is one of key node of interoception (Craing 2009), we hypothesized that resting functional connectivity from AI were involved in an effect of interoceptive training. To address this issue, we conducted a longitudinal intervention study using the interoceptive training (Suawara 2020), and obtained resting state fMRI before and after the intervention. Also, we evaluated interoceptive accuracy by using a heartbeat perception task (Schandry 1981). Twenty-two healthy volunteers (15 females, age 19.9 ± 2.0 years) participated, including graduate and undergraduate university students. The study protocol was approved by the ethical committee of NCNP. Informed consent was obtained from each subject. After the intervention, interoceptive accuracy was enhanced, and anxiety levels and somatic symptoms were reduced compared with baseline periods. Also, resting functional connectivity from AI to dorsolateral prefrontal cortex (DLPFC) , superior marginal gyrus (SMG), anterior cingulate cortex (ACC) and brain stem including nucleus tractus solitarius (NTS) were enhanced, and those from AI to visual cortex (VC) were decreased according to enhanced interoceptive accuracy due to the interoceptive training. The findings suggest that neural circuit of AI, ACC and NTS are involved in main pathway of interoception. The results indicate these networks are neural underpinnings of enhanced interoceptive accuracy due to training. Neural circuit of AI and DLPFC and SMG is thought to represent the central control process of regulation, by monitoring and manipulating emotion representations in the working memory (Kohn et al., 2014). Given that interoceptive attention was associated with reduced coupling between AI and VC (Wang 2019), our interoceptive training trains interoceptive perceptions, while changing the brain circuitry for exteroceptive perceptions. The findings provided a better understanding of an effect of interoceptive training on neural network of interoception as well as exteroception. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-037 情動視覚刺激に対する内側前頭前野ニューロンの応答性 Neuronal responsiveness of monkey ventromedial prefrontal neurons to emotional visual stimuli *戸塚 めぐみ(1)、岩沖 晴彦(2)、鴻池 菜保(2)、中村 克樹(2) 1. 京都大学大学院理学研究科、2. 京都大学霊長類研究所 *Megumi Totsuka(1), Haruhiko Iwaoki(2), Naho Konoike(2), Katsuki Nakamura(2) 1. Graduate School of Science, Kyoto University, 2. Primate Reserch Insititute, Kyoto University Keyword: Emotion, Ventromedial prefrontal cortex, Single-unit recording, Monkey The ventromedial portion of the prefrontal cortex (vmPFC) is a large heterogenous structure including areas 10, 14, 24, 25, and 32. Neuroimaging and neuropsychological studies have suggested that its dysfunction can induce mood and anxiety disorders. However, the relatively low spatial resolution of these studies failed to elucidate distinct functions of these heterogenous areas. To understand functional differences among these vmPFC regions, we previously examined the responsiveness of single neurons to various emotional visual stimuli in area 24 (Konoike et al. 2020). The present study focused on areas 14 and 25 because the functions of areas 14 and 25 are mostly unknown among the regions. We recorded the responsiveness of single neurons to various emotional visual stimuli in monkey areas 14 and 25, and directly compared response properties among areas 14, 24, and 25.Here, we used a passive viewing task. We prepared five categories of emotional stimulus: monkey faces, monkey hips, snakes, foods, and artificial objects. In this task, when the monkey pressed a lever and kept it pressed for a certain period (2.0 s – 3.0 s), a yellow fixation point (FP) appeared at the center of the monitor. If the monkey kept the lever pressed and fixated on the FP for 1.0 s, a visual stimulus was presented for 0.5s. Thereafter, the yellow FP was replaced by a red FP after a certain period (0.5 s – 1.5 s). If the monkey released the lever within 0.8 s of the replacement, the trial was regarded as correct and drop of water was delivered as a reward. We applied this task to two Japanese macaques (Macaca fuscata, male, 6.5-7.5 kg). We found some functional differences. In total, the activity of 209 neurons was recorded in the present study. Among these, 116 neurons (56 %) showed a significant change in activity in response to the emotional stimuli. The percentage of the responsive neurons was significantly higher than that in area 24 (46/174, 26%). Response latencies of the responsive neurons were longer than those in area 24. In addition, food, object, and snake stimuli elicited prompt responses in the present study whereas monkey and snake stimuli did in area 24. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-038 カルモジュリンキナーゼIIα活性と情動行動 Calmodulin kinase IIα activity and emotional behavior *山肩 葉子(1,2)、柳川 右千夫(3) 1. 自然科学研究機構生理学研究所、2. 総合研究大学院大学、3. 群馬大学大学院医学系研究科 *Yoko Yamagata(1,2), Yuchio Yanagawa(3) 1. Natl Inst for Physiol Sci, Okazaki, Japan, 2. SOKENDAI, Okazaki, Japan, 3. Grad Sch Med, Gunnma Univ, Maebashi, Japan Keyword: calmodulin kinase, kinase-dead, knock-in mouse, emotion Ca2+/calmodulin-dependent protein kinase IIα (CaMKIIα) is a key mediator of activity-dependent neuronal modifications and involved in the molecular mechanisms of learning and memory. It is also implicated in the regulation of emotional behavior. Previous studies reported that anxiety-related behaviors were reduced in the CaMKIIα-knock-out mouse and in the phosphorylation-deficient CaMKIIα (T286A)-knock-in (KI) mouse, while those behaviors were enhanced in the transgenic mouse overexpressing CaMKIIα in the forebrain. Thus, the level of CaMKIIα seems to be related to anxiety-related behaviors. On the other hand, we previously showed that the kinase-dead CaMKIIα (K42R)-KI mouse was severely impaired in hippocampus-dependent memory, and that the KI mouse was also affected in emotional behavior, as exemplified by decreased fear response after single training but generalized fear response after repeated training of fear conditioning. Here we performed other unconditioned response tests to evaluate more about emotional behavior of the kinase-dead CaMKIIα-KI mouse. In the open field locomotion test, the KI mouse showed increased locomotor activity especially at the beginning, indicating hyperactivity in a novel environment. The mouse also showed decreased time spent in the center area and increased traveling distance in the peripheral area, indicating the avoidance of an open space. In the elevated plus maze test, the KI mouse showed increased entry and time spent in the open arms, indicating the preference for an open space. Based on these and other behavioral test results, we will discuss the emotional traits in the absence of kinase activity of CaMKIIα. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-039 Fear-memory-associated neurons in ventral hippocampus may promote arousal from sleep *Yi-Tse Hsiao(1), Ting-Yen Lee(1), Ching-Yuan Chang(1) 1. Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei, Taiwan Keyword: fear memory, sleep, hippocampus, amygdala Although fear leads to unpleasant feelings, it helps animals to avoid potential risks. After experiencing fearful events, sleep problems such as nightmares frequently occur. Studies have reported that ventral hippocampus and amygdala modulate stress relative behaviors: memorizing fear events, for example. In this study, we hypothesized that footshock stimulation causes fear memories to encode in neurons of the ventral hippocampus which projects to the basal amygdala. In addition, activation of these neurons results in sleep problems. To test our hypothesis, we used a miniature microscope and also fiber photometry to observe the neuronal activity of ventral CA1 (vCA1) and compared their calcium activity during sleep post to footshock stimulation. We found the activities of vCA1 were increased and the cell-cell interaction enhanced when they heard conditioned stimuli during sleep after fear conditioning. We then used the activity-dependent labeling technique to express channelrhodopsin on fear-memory-relative neurons in the vCA1 and photostimulated the amygdala. The data shows that activation of the vCA1-amygdala fear memory relative neurons arouses the sleeping mice. Our data suggests that vCA1 to amygdala pathway exists in neurons that encode fear memories and activation of these neurons disrupts sleep. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-040 初発、無投薬うつ病患者の臨床的特徴と扁桃体亜区域容積との関連 Volume of amygdala subregions and clinical manifestations in patients with first-episode, drug-naïve major depression *手銭 宏文(1)、渡邉 啓太(2)、岡本 直通(1)、池ノ内 篤子(1)、井形 亮平(1)、小西 勇輝(1)、掛田 伸吾(3)、吉村 玲児(1) 1. 産業医科大学　精神医学、2. 京都大学　オープンイノベーション機構、3. 弘前大学大学院医学研究科　放射線診断学講座 *Hirofumi Tesen(1), Keita Watanabe(2), Naomichi Okamoto(1), Atsuko Ikenouchi(1), Ryohei Igata(1), Yuki Konishi(1), Shingo Kakeda(3), Reiji Yoshimura(1) 1. Department of psychiatry, University of Occupational and Environmental Health, Kitakyushu, Japan, 2. Open Innovation Institute, Kyoto University, Kyoto, Japan , 3. Department of Radiology, Graduate School of Medicine, Hirosaki University, Hirosaki, Japan Keyword: amygdala, subregions, major depression, MRI Although the pathophysiology of major depression (MD) remains unclear, recent brain imaging studies using MRI have provided some interesting insights. Previous studies of the amygdala and hippocampus have reported reduced volume as a potential biomarker of depression. However, few information about amygdala volume for MD patients. There have been also few reports on the relationship between MD and amygdala subregional volume. One of the reasons for this is that the amygdala nuclei have been difficult to classify and identify. Additionally, the amygdala has different functions depending on its region. It is important to investigate the relationship between the amygdala subregional volume and clinical symptoms of MD, which might be shed light on the role of subregions of amygdala for pathophysiology of MD. 　The aim of the present study is to investigate the amygdala subregion volumes in patients with a first episode of MD and in healthy subjects. Covariate-adjusted linear regression was performed to compare the MD and healthy groups, and adjustments for age, gender, and total estimated intracranial volume showed no differences in amygdala subregion volumes between the healthy and MD groups. Within the MD group, we examined the association between amygdala subregion volume and the 17-item Hamilton Rating Scale for Depression (HAMD) score and the HAMD subscale score. There was an inverse linear association between the HAMD total and the HAMD core and lateral nucleus and anterior-amygdaloid-regions, but no associations in the left amygdala. Furthermore, an inverse linear association was seen between the HAMD psychic and the lateral nucleus, anterior-amygdaloid-regions, transition, and whole amygdala. 　The findings of this study suggest that the severity of MD and some depressive symptoms are associated with the subregions of right amygdala volume. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-041 慢性ストレスへのミクログリア応答の左右非対称性 Left-right asymmetry in chronic stress-induced microglial responses *松下 和敏(1)、谷口 将之(1)、古屋敷 智之(1) 1. 神戸大学大学院・医学研究科・薬理学分野 *Kazutoshi Matsushita(1), Masayuki Taniguchi(1), Tomoyuki Furuyashiki(1) 1. Div. Pharmacol., Grad. Sch. Med., Kobe Univ. Keyword: microglia, chronic stress Chronic social stress may cause emotional alterations including depression and increased anxiety and is a risk factor for psychiatric disorders. In mice, chronic social stress activates microglia in the medial prefrontal cortex (mPFC) through innate immune receptors TLR2/4, leading to neuronal dysfunctions and social avoidance. Since lipopolysaccharide, a TLR4 ligand, differently induces cytokine production from microglia isolated from the left and right hemispheres, we speculate that chronic stress induces asymmetric microglial responses in the mPFC. Here we optimized the procedure of transcriptome analysis of subregion-specific mPFC microglia isolated by laser microdissection. Using this technique, we analyzed chronic social stress-induced gene expression changes in mPFC microglia of left and right hemispheres. Given individual variability in stress susceptibility, we categorized the stressed mice into susceptible and resilient mice based on the level of social avoidance. We identified genes that were up- or down-regulated by chronic social stress in left and right mPFC. These genes showed differential responses to the chronic stress regarding the left-right asymmetry and individual variability of stress susceptibility. Notably, the genes that were upregulated specifically in resilient mice coincided with those with the left-right asymmetry. These findings suggest the presence of left-right asymmetry of chronic stress-induced neuroinflammation and its relevance to stress resilience. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-042 メダカをモデルとした行動に関わる性格の分子神経基盤解析 Analysis of the molecular and neural basis of animal behavioral personality in medaka *田中 豪(1)、成瀬 清(2)、郷 康広(3)、辰本 将司(3)、豊田 敦(4)、勝村 啓史(5)、中川 真一(1)、横井 佐織(1) 1. 北海道大学大学院薬学研究院、2. 基礎生物学研究所、3. 自然科学研究機構、4. 国立遺伝学研究所、5. 北里大学 医学部解剖学 *Tsuyoshi Tanaka(1), Kiyoshi Naruse(2), Yasuhiro Go(3), Shoji Tatsumoto(3), Atsushi Toyoda(4), Takafumi Katsumura(5), Shinichi Nakagawa(1), Saori Yokoi(1) 1. Faculty of Pharmaceutical Sciences, Hokkaido University, 2. National Institute for Basic Biology, 3. National Institutes of Natural Sciences, 4. National Institute of Genetics, 5. Department of Anatomy Kitasato University School of Medicine Keyword: animal personality, GWAS analysis, boldness/shyness, medaka Two traits of "boldness" and "shyness" have been studied as part of animal personality for a long time, but the genes that are responsible for these traits have not yet been identified. In this study, we performed the anxiety-like behavior test and quantified boldness/shyness personalities of several novel closed colonies (Matings have occurred within the colony members randomly for a long time, resulting in limited genetic diversity) derived from wild medaka (Oryzias latipes) in the West Setouchi region. Among them, we found a closed colony with high shyness and that with high boldness (“bold colony” and “shy colony”). Since d-rR strain, one of the standard experimental strains, showed intermediate traits between these two strains, it is possible that the genes that are responsible for boldness of medaka can be newly identified by these novel closed colonies. We compared the gene expression levels in the brains of the bold and shy colonies and narrowed down the responsible gene. In addition, we prepared about 150 F2 individuals, which were crossed between the bold and shy F1 colonies and extracted their genomes from their tails after the behavioral experiments. We selected the top 27 bold fish and the top 27 shy fish out of the 150 and searched for the genomic regions responsible for the difference between the bold and shy colonies by GWAS (Genome Wide Association Study) using their entire genome sequences. Now, in order to examine whether neural responses of bold and shy fish towards anxiety stimulus are different, we are performing whole mount in situ hybridization experiments targeting egr1 used as an indirect marker of neuronal activity. Next, we plan to edit candidate genes and genome regions of these colonies to analyze the molecular and neural basis of boldness. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-043 不安様行動時における前障神経細胞のカルシウムイメージング Calcium imaging of claustral neurons during anxiety-like behaviors *田沼 将人(1)、笠井 淳司(1)、宮司 渓汰(1)、大久保 仁(1)、橋本 均(1,2,3,4,5) 1. 大阪大院・薬・神経薬理、2. 大阪大院・連合小児、3. 大阪大・データビリティフロンティア機構、4. 大阪大・先導的学際研究機構、5. 大阪大・院医・分子医薬 *Masato Tanuma(1), Atsushi Kasai(1), Keita Miyaji(1), Jin Ohkubo(1), Hitoshi Hashimoto(1,2,3,4,5) 1. Lab. Mol. Neuropharmacol., Grad. Sch. Pharmaceut. Sci., Osaka Univ., Osaka, Japan, 2. United Grad. Sch. Child Dev., Osaka Univ., Osaka, Japan, 3. Inst. Datability Sci., Osaka Univ., Osaka, Japan, 4. Inst. for Open Transdisciplinary Res. Initiatives, Osaka Univ., Osaka, Japan, 5. Dept. of Mol. Pharmaceut. Sci., Grad. Sch. of Med., Osaka Univ., Osaka, Japan Keyword: Claustrum, Calcium Imaging, Anxiety We have recently reported that excitatory neurons in the claustrum mediate responses to acute psychological stressors that induce negative emotional states and anxiety-like behaviors. However, the neuronal representations that underlie such states and behaviors are not well understood. To examine the activity of claustral neurons during these behaviors, here we performed freely moving calcium imaging of excitatory neurons in the claustrum that express the genetically encoded calcium indicator GCaMP6f under the CaMKIIα promoter. The in vivo neuronal activity of a mouse was recorded from three behavioral tests; the elevated plus maze, the open field test, and a second open field test after an exposure to a ten-minute single social defeat stress. We found that a subset of claustral neurons displayed an increase in the rate of calcium events when in the open arm of the elevated plus maze. In addition, claustral neurons showed an increase in calcium levels upon transitioning from the closed arm into the open arm. Consistent with these results, claustral neurons also showed an increase in calcium levels upon exiting the corner zones of the open field. Exposure to a stressor, however, rendered the calciums levels of a subset of claustral neurons to remain high in the corner zones of the open field. Post hoc tracking of single neurons across the behavioral tests showed that approximately 50% of this subset of neurons were those that showed increased calcium levels upon entry into the open arm in the elevated plus maze. These results suggest that negative emotional states and anxiety-like behaviors are encoded by a subset of neurons in the claustrum. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-044 Anxiety-like behaviors in rats exposed to traffic-related air pollutants in early life *Tin Tin Win Shwe(1), Chaw Kyi Tha Thu(2,3), Yuji Fujitani(1), Shinji Tsukahara(2), Seishiro Hirano(1) 1. National Institute for Environmental Studies, Tsukuba, Japan, 2. Division of Life Science, Graduate School of Science and Engineering, Saitama University, Saitama City, Japan, 3. Department of Immunology, International University of Health and Welfare, School of Medicine, Narita, Japan Keyword: Anxiety, Developmental exposure, Diesel exhaust, Neurotransmitters Background and Aim: Autism spectrum disorder (ASD) is a complex neurodevelopmental disorder and both genetic and environmental factors are potential causative factors for ASD. It was reported that 40% of children and adolescents with ASD are at increased risk of anxiety and anxiety disorders. Recently, we have reported that developmental exposure to diesel exhaust origin secondary organic aerosol (DE-SOA) is associated with cognitive deficit and autism-like behaviors in rat offspring. However, the effects of DE-SOA on anxiety and its mechanism are not well-understood. We aimed to examine the effects of developmental exposure to DE or DE-SOA on anxiety-like behaviors and neurotransmitters, neurotrophic factors, and inflammatory markers in the rat offspring. Methods: Sprague-Dawley pregnant rats were exposed to clean air (control), DE (100 mg/m3) and DE-SOA (120 mg/m3) in the inhalation chamber from gestational day 14 to postnatal day 21. On 7 to 9-week-old, anxiety-like behaviors such as open field test (OFT), elevated plus maze (EPM), light/dark test (LDT) and novelty-induced hypophagia (NIH) were performed. Then, the hippocampus was collected to examine neurological and immunological markers by real-time RT-PCR method and immunohistochemical analysis. Results: Among behavior tests, OPF, EPM and NIH showed anxiety-like behaviors in both male and female rat offspring exposed to DE or DE-SOA. Serotonin receptor (5HT1A), dopamine receptor (D2R), brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor A (VEGFA) mRNAs were significantly decreased and interleukin (IL)1 b, cyclooxygenase (COX) 2 and heme oxygenase (HO) 1 mRNAs were significantly increased in the hippocampus of both male and female rats exposed to DE or DE-SOA. Discussion and Conclusion: Our results indicate that developmental exposure to air pollutants may induce anxiety-like behaviors via modulation of neurological and immunological markers in rats. We suggest that neuroinflammation, neurodegeneration and oxidative stress are possible mechanism of air pollutant-induced anxiety-like behaviors in autistic subjects. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-045 視床室傍核の遺伝子発現と神経投射の明瞭な差と連続性 Distinctive and continuous transcriptome and connectivity in the anterior-posterior axis of the paraventricular nucleus of thalamus *島 康之(1)、Henrik Skibbe(1)、二階堂 愛(4)、服部 信孝(2)、加藤 忠史(3) 1. 理化学研究所　脳神経科学研究センター、2. 順天堂大学　大学院医学研究科　脳神経内科、3. 順天堂大学　大学院医学研究科　精神医学講座、4. 東京医科歯科大学　難治疾患研究所 *Yasuyuki Shima(1), Henrik Skibbe(1), Itoshi Nikaido(4), Nobutaka Hattori(2), Tadafumi Kato(3) 1. RIKEN CBS, 2. Department of Neurology, Jyuntendo University, 3. Department of Psychiatry, Jyuntendo University, 4. Department of Functional Genome Informatics, Tokyo Medical and Dental University Keyword: scRNAseq, cell type, PVT The paraventricular nucleus of the thalamus (PVT) projects axons to multiple brain areas and mediates a wide range of behaviors related to motivated behaviors, emotion, and saliency. Regional differences of function and axonal projection in PVT, especially between anterior and posterior PVT, have been reported, but what cell types exist in PVT and how different they are anatomically and functionally have not been addressed. We applied single-cell RNA sequencing to depict transcriptomic characteristics of mouse PVT neurons. The transcriptome of PVT neurons had a continuous distribution with the largest variance corresponding to the anterior-posterior axis. Although the single-cell transcriptome classified PVT neurons into 4 types, intermediate population existed between any pair of cell types. Multicolor fluorescent in sit and signal quantification revealed that distinct expression of cell type marker genes were observed at either anterior- and posterior-ends while in-between had many marker-negative cells, supporting continuity in the transcriptome of PVT. We also analyzed whole-brain axonal projection of PVT subpopulation using Cre line with labeling specific populations. Subpopulations of aPVT and pPVT had nearly non-overlapping axon projection patterns, while another population showed intermediate patterns. In addition, the subpopulations responded to Orexin A (OrxA) and γmelanocortin-stimulating hormone (MSH), and their chemogenetic activation showed opposing effects in food consumption. Our studies showed contrasts and continuity of PVT neurons underlying their function as a behavior-modulating hub. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-046 扁桃体基底外側部に存在するニューロペプチドYニューロンの解剖学的・機能的解析 Anatomical and functional analysis of neuropeptide Y neurons in the basolateral amygdala *山田 俊児(1)、渡邊 義久(2)、田中 雅樹(1) 1. 京都府立医科大学大学院医学研究科　生体構造科学、2. 京都府立医科大学大学院医学研究科　基礎老化学 *Shunji Yamada(1), Yoshihisa Watanabe(2), Masaki Tanaka(1) 1. Department of Anatomy and Neurobiology, Grad Sch Med, Kyoto Prefectural Univ of Med, 2. Department of Basic Geriatrics, Grad Sch Med, Kyoto Prefectural Univ of Med Keyword: Neuropeptide Y, basolateral amygdala, nucleus accumbens, retrograde AAV Neuropeptide Y (NPY) is a 36-amino acid neuropeptide that is widely expressed in the central nervous system, including the cerebral cortex, hypothalamus, and amygdala. We previously suggested that NPY-expressing neurons in the nucleus accumbens (NAc) modulate anxiety-behavior and these neurons project to the lateral hypothalamus using NPY-Cre mice and Cre-dependent AAV. Moreover, we also found that the origin of some NPY fibers in the NAc is the basolateral amygdala (BLA). Obtaining more knowledge about BLA NPY neurons, in the present study, we investigated anatomical characteristics and functions of BLA NPY neurons. To investigate projection of BLA NPY neurons, we injected AAV(DJ)-FLEX-mGFP-t2A-synaptophysin-mRuby into the BLA in NPY-Cre mice. We found many GFP-positive fibers in the NAc, anterior cingulate cortex (ACC), and bed nucleus of the stria terminalis (BNST). Injection of retrogradely AAV(rg)-FLEX-mCherry into three regions caused expression of mCherry-positive cells in the BLA, suggesting that BLA NPY neurons project to the NAc, ACC, and BNST. Next, we performed region-specific ablation of BLA NPY neurons using AAV(DJ)-FLEX-taCasp3 and investigated the effect of BLA NPY neurons on the food intake and anxiety behaviors in the normal or acute stress condition. Analysis of in situ hybridization revealed that many NPY mRNA-expressing cells found in the BLA of control mice, while only a few NPY mRNA-expressing cells were found in AAV(DJ)-FLEX-taCasp3-treated mice. The ablation of NPY neurons in the BLA did not affect to the 24-h food intake and anxiety behaviors in normal conditions. However, in acute stress (after 30 min or 120 min immobilization stress) condition, the ablation of BLA NPY neurons caused reduction of the percentage of time spent in the open arms in the elevated plus maze test and the percentage of time spent in the central area in the open field test, but not in control mice. These results indicated that NPY neurons in the BLA may be involved in attenuation of anxiety in stressful condition in mice. Further studies are needed in order to clarify the projection site (s) of BLA NPY neurons mediating attenuation of anxiety after acute stress. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-047 lncRNA TUNAR 内の非古典的ORF欠損マウスはプレパルス抑制の増強とうつ関連行動の増加を示す Loss of non-canonical open reading frame within lncRNA TUNAR increased pre-pulse inhibition and depression-related behavior in mice *藤井 一希(1,2)、森脇 雄介(3)、腰高 由美恵(2)、安達 真由美(2)、柳橋 裕子(2)、本郷 翔子(2)、相澤 康則(4)、高雄 啓三(1,2,3) 1. 富山大学 学術研究部医学系 行動生理学講座、2. 富山大学 研究推進機構 研究推進総合支援センター 生命科学先端研究支援ユニット、3. 富山大学 医学薬学教育部 行動生理学講座、4. 東京工業大学 生命理工学院 *Kazuki Fujii(1,2), Yusuke Moriwaki(3), Yumie Koshidaka(2), Mayumi Adachi(2), Yuko Yanagibashi(2), Shoko Hongo(2), Yasunori Aizawa(4), Keizo Takao(1,2,3) 1. Dept. Behav. Physiol., Fac. Med., Univ. Toyama, Toyama, Japan, 2. Life Sci Res Ctr, Univ. Toyama, Toyama, Japan, 3. Dept. Behav. Physiol., Grad. Sch. Med. Pharma. Sci. Edu., Univ. Toyama, Toyama, Japan, 4. Grad. Sch. Biosci. Biotech., Tokyo Institute of Technology, Yokohama, Japan Keyword: microprotein, behavioral test battery, depression, non-canonical ORF By definition, long noncoding RNAs (lncRNAs) do not contain protein-coding open reading frames (ORFs). Recent bioinformatics and high-throughput sequencing studies, however, reported that many lncRNAs possess short “non-canonical” ORFs (sORFs) encoding microproteins. In particular, Ribo-seq, a technology using deep-sequencing to identify the positions of ribosomes engaged in translation, revealed that lncRNA contains many sORFs previously considered to be untranslatable. These newly identified microproteins are expected to be novel factors that will contribute to our understanding of life science and are potential targets of drug discovery. The Tcl1 upstream neuron-associated RNA (TUNAR) was discovered as a lncRNA. The TUNAR sequence is remarkably conserved across vertebrates and is highly expressed in neural tissues. By in silico screening, we identified that TUNAR has a sORF region encoding a 48-amino acid polypeptide. The lncRNA TUNAR plays a vital role in pluripotency and neural differentiation of mouse embryonic stem cells. Whether or not the sORF is translated into a microprotein in vivo, and how TUNAR and the microprotein affect brain function and behaviors, however, remain unclear. Here, we generated TUNAR-microprotein reporter mice and TUNAR sORF deletion mice on the C57BL/6J background using the CRISPR-Cas9 genome editing system. Utilizing TUNAR-microprotein reporter mice in which an epitope-tag coding sequence was inserted before the stop codon, we detected the epitope tag-specific stained cells in the mouse central nervous system (e.g., thalamus, inferior colliculus, pons). These findings suggested that the TUNAR sORF is translated into an endogenous protein. To investigate the function of TUNAR microprotein in the brain, we subjected TUNAR sORF deletion mice to a comprehensive behavioral test battery. TUNAR sORF deletion mice had significantly lower body weight than their wild-type (WT) littermates. In the rotarod test, TUNAR sORF deletion mice exhibited better motor coordination compared with WT mice. In the startle response/pre-pulse inhibition (PPI) test, while there was no significant difference in the acoustic startle response, TUNAR sORF deletion mice showed increased PPI compared with WT mice. In both the Porsolt forced swim test and tail suspension test, TUNAR sORF deletion mice exhibited increased depression-related behavior. These results suggest that the TUNAR sORF deletion induces a depressive effect. These findings together indicate that the non-canonical microprotein from TUNAR is translated in the brain and has a critical role in sensory-motor gating and depression-related behavior. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-048 反復挫折ストレスに誘導されたうつ様行動におけるオートファジーの分子機構の解明 Deficient autophagy in microglia aggravates repeated social defeat stress-induced social avoidance *坂井 舞(1)、兪 志前(1)、小野 千晶(1)、菊地 淑恵(1)、富田 博秋(1) 1. 東北大学 *Mai Sakai(1), Zhiqian Yu(1), Chiaki Ono(1), Yoshie Kikuchi(1), Hiroaki Tomita(1) 1. Tohoku University Keyword: AUTOPHAGY, MICROGLIA, STRESS Autophagy signaling carries its components into the intracellular digestive system and lysosomes and degrades them to promote survival. Autophagy is activated under various stress conditions that are associated with several diseases in the brain. Microglia are major immune cells in the central nervous system, which involve in stress is evident, including changes in microglial density in depression patients. This study aimed to clarify the autophagy signaling changes in the prefrontal cortex under repeated social defeat and investigate the involvement of microglial autophagy in stress-induced behavioral changes. We found that repeated social defeat stress, an animal model of depression, significantly induced initial autophagic signals followed by increased transcription of autophagy-related genes in the PFC. Similarly, significantly increased transcripts of ATGs (Atg6, Atg7, Atg12, and Atg5) were confirmed in the postmortem PFC of patients with MDD. The protein levels of the prefrontal cortical LC3B were significantly increased, whereas p62 was significantly decreased in the resilient but not in susceptible mice. The above results indicate that enhanced autophagic flux may alleviate stress-induced depressive symptoms. Furthermore, we identified that an early-stage autophagy regulator, FKBP5, was significantly increased in the PFC of resilient mice at the transcript and protein levels. In addition, the resilient mice exhibited enhanced autophagic flux in the prefrontal cortical microglia, and the autophagic deficiency in microglia aggravated RSD-induced social avoidance, indicating that microglial autophagy involves stress-induced behavioral changes. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-049 C57BL/6マウス亜系統間の違い：異なる行動特性を引き起こすマイクロ回路基盤の探索 C57BL/6 substrains are different: Exploration of causal microcircuit mechanisms of distinct behavioral traits *加瀬田 晃大(1)、Bolati Wulaer(1)、長井 淳(1) 1. 理化学研究所 脳神経科学研究センター グリア−神経回路動態研究チーム *Kodai Kaseda(1), Bolati Wulaer(1), Jun Nagai(1) 1. Laboratory for Glia-Neuron Circuit Dynamics, RIKEN Center for Brain Science, Wako, Japan Keyword: Mouse strain, Behavior, Monoamine, c-Fos Behavioral traits in responses to chemical, physical or environmental factor differ among individuals, that is at least partially derived from genetic factors. Commercially available experimental mouse strains, including C57BL/6 mouse line, are known to vary by gene expressions and behave differently. However, its circuit and microcircuit mechanisms that fill the gap between genes and behavior remain elusive. Here we tackled this problem with C57BL/6 mouse line “sub”strains, using monoamine analysis, psychostimulant responses, behavioral battery, and c-Fos mapping. We found that substrains exhibit brain nuclei-specific monoamine levels and distinct exploratory, alertness-related behaviors. Using high performance liquid chromatography, the monoamine levels were different in the frontal and limbic areas between the substrains. In accord with this, methamphetamine, an inhibitor of monoamine transporter, induced distinct hyper-locomotive activity. Furthermore, anxiety-related behavior, e.g. time spent in the center of an open field, a lighted arena and open arms of elevated plus maze varied between substrains. Mice from substrains also showed difference in encoding, retrieval and generalization of conditioned fear. To tease apart circuit and cellular basis of these results, we report c-Fos mapping of neurons and astrocytes across brain regions in combination with behavioral assays. Our data suggest that C57BL/6 substrains differ in behavioral traits in responses to chemical and/or environmental factors. Their distinct monoamine levels in the brain may implicate individual’s difference in responses to illicit drugs. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-050 Violet light treatment ameliorates social behaviour in social defeat stress mice *Pooja Gusain(1), Sasaki Nobunari(1), Yasue Mitsukura(3), Motoshi Hayano(1,2,3,4), Kazuki Tsubota(1,4) 1. Department of Ophthalmology, Keio University, School of Medicine, Tokyo, Japan , 2. Department of Neuropsychiatry, Keio University, School of Medicine, Tokyo, Japan , 3. Faculty of Science and Technology, Keio University, Yokohama, Japan , 4. Tsubota Laboratory, Inc., Tokyo, Japan Keyword: Mood, Violet light, Social defeat stress , Oligodendrocytes Lack of necessary light exposure could significantly give rise to the behavior pattern like mood disorders especially the depression. In this mechanism, the stimulation of light via non-visual system photoreceptors affects various functions of the brain. Therefore, these photoreceptors are vital in everyday life such as colour vision. Previous research evidently supports the light therapy as an efficient treatment in depression and memory functions. Recent study has also shown that violet light (VL) (360-400nm wavelength) can activate OPN5 photoreceptor which is important for eyes as well as neuro-linked brain functions. Violet light is triggered in the retina or skin via OPN5 and significantly affects sleep, memory, emotions, body temperature, etc. In recent days, Violet light emerges as potential light therapy in eye research, however exact effects and mechanisms is still to be determined in mood disorder. In our research model, we hypothesize that VL driven oligodendrocytes modulation ameliorates depressive behaviour through neuronal activation in nucleus accumbens(NAC)- paraventricular thalamus(PVT) region. In this study, we examine the effect of violet light exposure on the regulation of oligodendrocytes brain activity in social defeat stress (SDS) depression model. Briefly, C57B6/J mice were subjected to aggressor mice (ICR) to a defeat session to induce depression. After each defeat session mice were separated by perplex glass to allow sensory and visual threat till next defeat session. The stressed susceptible mice were then exposed to violet light followed by behavioural test and molecular analysis. The behavioral results indicate that in SDS mice, VL exposure renders antidepressant like effect while no significant effect was observed to white light (WL). Also, the defeated mice exposed to VL shows increase social interaction ratio and less time spent in the corner zone compared to the defeated mice exposed to WL. In addition, c-fos neural activation under VL in SDS mice revealed expression in distinct brain regions including NAc and PVT. Moreover, VL exposed SDS mice shows significant increase in the MBP mylination compared to WL defeated mice. VL provides significant effect as a novel non-invasive treatment in regulating mood disorders as a potential antidepressant. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-051 The role of D1- and D2-type dopamine receptors in Central Amygdala in processing cocaine-related experiences *Lukasz Bijoch(1), Leszek Kaczmarek(2), Anna Beroun(1) 1. Laboratory of Neuronal Plascticity, BRAINCITY, Nencki Institute of Experimental Biology PAS, Warsaw, Poland, 2. Laboratory of Neurobiology, BRAINCITY, Nencki Institute of Experimental Biology PAS, Warsaw, Poland Keyword: Addiction, Dopamine, Amygdala, Reward Cocaine is one of the most reinforcing drugs-of-abuse, which drives awareness, alertness and a neurochemical magnification of pleasure. Despite its physical harmfulness, cocaine’s powerful addictive potential makes it the second most popular recreational drug-of-abuse in Europe. Cocaine causes enjoyable sensations because it potentiates dopamine release in the brain reward system. Moreover, by inhibiting dopamine transporter (DAT) cocaine increases extracellular concentration of dopamine, leading to neuroplastic changes of dopaminoceptive neurons (cells which express dopamine receptors). There are two major types of dopamine receptors, which exert opposing actions: D1-type (D1DR, increasing intracellular signal transduction) and D2-type (D2DR, decreasing intracellular signal transduction). Recently, we found highly non-overlapping populations expressing these receptors in two subunits of Central Amygdala. Neurons with either D1DR or D2DR in its medial part (CeM) and cells with solely D2DR in lateral (CeL). As there are increasing number of evidences that Central Amygdala is crucial for processing appetitive (i.e., of positive valence) experiences, we studied these cells in the context of addictive reward exposure. To study different dopaminoceptive neuronal populations, transgenic mice with fluorescently labeled D1DR- or D2DR-type neurons were used. Mice underwent 7 days of either cocaine or saline intraperitoneal injections. Two hours after the last reward exposure, mice were sacrificed and whole cell patch clamp experiments were performed on neurons in CeM and CeL. All recordings were performed with contradistinction of D1DR and D2DR-type neurons. Biocytin was added to the internal solution for later visualizing morphology of patched cells. We found opposite adaptations of dopaminoceptive neurons in CeM. There, upon cocaine exposure, D2DR positive neurons increase their spontaneous activity while D1DR positive diminish it. Also, we observed in CeM amplified excitability of D2DR neurons and reduced in the case of D1DR cells. These electrophysiological adaptations went alongside with changes in morphology of these cells: variations in the number and volume of their dendritic spines. Surprisingly, D2DR-positive neurons in CeL exhibited opposite adaptations to these in CeM. There, we found that cocaine exposure caused reduction of neuronal spontaneous activity and excitability. Our results show that CeA circuitry appears to be a complex player in the drug-context associations. Moreover, adaptations of dopaminoceptive neurons in CeM seems to be opposite to these observed in other parts of the brain, e.g. nucleus accumbens. Thus, the role of D1DR- and D2DR-type neurons in reward processing cannot be generalized. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-052 嗜癖行動を想起する環境刺激情報処理メカニズム Environmental Cue Processing Associated with Behavioral Addiction *浅岡 由衣(1,2)、元 武俊(3)、石川 恵己(4)、森田 智也(4)、後藤 幸織(1) 1. 京都大学霊長類研究所、2. 日本学術振興会特別研究員DC、3. MRCラボクリニック、4. 特定医療法人共和会 共和病院 *Yui Asaoka(1,2), Moojun Won(3), Emi Ishikawa(4), Tomonari Morita(4), Yukiori Goto(1) 1. Primate Res Inst, Kyoto Univ, Inuyama, Aichi, 2. JSPS Research Fellowship for Young Scientists, 3. MRC Lab Clinic, Mitaka, Tokyo, 4. Kyowa Hospital, Obu, Aichi Keyword: Addiction, Eye tracking, NIRS, Prefrontal cortex Addiction to a particular activity, such as gambling, Internet use, and gaming, is problematic and subject to treatment. Some impulse control disorders, such as kleptomania and paraphilia, are also thought to meet the criteria of addiction. However, insufficient studies still make it difficult to comprehensively understand these disorders as a behavioral addiction, and further investigation is required. To further elucidate insights about behavioral addiction, we investigated how environmental cues associated with the disorder were processed in patients with kleptomania in this study. The images of the grocery store with (MKT+H) and without (MKT-H) a person, those of outside with (OUT+H) and without (OUT-H) a person, and those of foods (FOD) and stationary (STY) were presented to healthy subjects and patients, who had been hospitalized for treatment of kleptomania for stealing foods in grocery stores, and their gazing patterns and prefrontal cortical (PFC) activity were measured with eye-tracking and functional near infrared-spectroscopy. Preliminary data analysis has unveiled no significant difference in any of the fixation patterns (number, duration, and dispersion), the number of blinks, and changes (coefficient of variance) of pupil diameter sizes between the patients and healthy subjects while gazing at each image. However, the number of fixations on the area of interest set to the person in the images of MKT+H was significantly lower in the patients and the healthy subjects, but such difference was not present in OUT+H. PFC hemoglobin dynamics were also not substantially different between the patients and healthy subjects, but correlation analysis found that PFC activity was highly correlated across all images in healthy subjects, whereas the PFC activity while gazing at the MKT-H image was significantly different from other images in the patients. These results suggest that, similar to drug addiction patients, patients with kleptomania exhibit distinct neuronal and behavioral responses to the environmental cues provoking their addicted behavior. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-053 Exploring Sex-Differences and a Role for the 5-HT System in Early Life Stress-Induced Compulsive Ethanol Consumption in Adulthood *Bryan D McElroy(1), Chen Li(1), Lynn G Kirby(1) 1. Temple University Lewis Katz School of Medicine, Philadelphia, USA Keyword: Sex-Differences, Alcohol Use Disorder, Early Life Stress, Serotonin Heavy drinking has declined in most industrialized nations but in Japan it is 4-6x higher than in 1960. ≈70% of Japanese are unaware of their problematic drinking, those with alcohol use disorder are considered weak willed which is reflected by only 5% seeking treatment. Japan has a reputation for its work ethic and in turn high tolerance for drinking to unwind highlighted by 60% of businessmen reporting heavy drinking being accepted as a part of their job and a mark of company loyalty leading to a high prevalence of nomisugi or 'too drunk' businessmen sleeping on the streets. Stress-vulnerable communities such as these, are more motivated to drink to 'self-medicate' their high stress lifestyles. The dorsal raphe nucleus (DRN)-serotonin (5-HT) system is sexually dimorphic, vulnerable to stress, important in stress-reward regulation and the ideal target to treat compulsive (despite negative consequences) drinking. We hypothesized that DRN 5-HT neurotransmission underlies the sex-dependent effects of stress on ethanol (EtOH) intake and compulsive drinking. Male and female Tph2-iCre rats or wild-type Sprague-Dawley littermates underwent social isolation stress (SIS) or group housing in adolescence, intraDRN viral delivery of Cre-dependent excitatory Gq-DREADDs (AAV-hSyn-DIO-hM3Dq-mCherry) and tamoxifen induction. They were exposed to 20% EtOH in a 3-week intermittent-access two-bottle choice (IA2BC) model. Next, transferred to a 10-day oral EtOH self-administration (SA) model and 3 days of punished SA (50% of EtOH deliveries + 0.24mA footshock) to test compulsive drinking. Clozapine-N-oxide was administered 30 mins prior to each punishment session to activate Gq-DREADDs. Results showed a sex x stress interaction in IA2BC EtOH intake. SA drinking revealed a time x stress interaction in males (higher drinking in SIS). No effect of SIS but females had higher SA EtOH intake than males and more shock-resistant (compulsive) drinking. Chemogenetic activation of DRN 5-HT neurons blunted punishment suppression of SA. Results suggest females are more sensitive to early life stress-induced rise in EtOH intake and adult compulsive drinking. Prior studies in our lab support stress-induced suppression of the DRN 5-HT system and chemogenetic activation of 5-HT appears to buffer these stress effects. In conclusion, we show the important effects of sex and stress on drinking and motivation for EtOH and a novel role of the 5-HT system in compulsive drinking. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-054 Neurochemical and behavioural differences between ketamine isomers and amphetamine in male rats *Shuo-Fu Chen(1,2), Chuan-Yao Wang(2), Chih-Chang Chao(2), Wei-Chung Hsu(1,3), Lei-Chieh Chao(1,3), Syun-Ruei Lee(3), Minpheus Huang(4), Ruey-Ming Liao(1,2) 1. Department of Psychology, National Cheng-Chi University, Taipei, Taiwan, 2. Institute of Neuroscience, National Cheng-Chi University, Taipei, Taiwan, 3. Brain Science Program, National Cheng-Chi University, Taipei, Taiwan, 4. Ohio State University, Columbus, OH, USA Keyword: drug abuse, behavioral sensitization, conditioned place preference, dopamine and glutamate Ketamine is repurposed in depression therapy, albeit the underlying neurobehavioral mechanisms remain elusive. Whether the psychostimulant-like effects of ketamine are associated to drug use or abuse is still a doubtful issue. Previous studies frequently used an injectable ketamine solution testing its rewarding effects, whereas comparatively fewer studies have explored ketamine isomer(s) on this issue. We investigated the effects of RS (+/-)-ketamine (RS-KET) and S (-)-ketamine (S-KET) on a behavioral protocol combining conditioned place preference (CPP) and behavioral sensitization (BS) paradigms. Additionally, d-amphetamine (d-AMP) treatment was conducted as a positive control. In a 30-day test, the CPP was conducted by either the unbiased or the biased procedure; and, BS test were run across three phases including a pre-test and a post-test conducted respectively before and after the development phase (7 days of drug injection intermittently separated by saline injects). The dose given at the pre- and post-tests was lower (i.e., a half) than that administered in the development phase. Behaviorally, in contrast to that d-AMP (1 mg/kg) significantly produced CPP, neither RS-KET (10 mg/kg) nor S-KET (10 mg/kg) produced CPP. However, a CPP was detected in the long-lasting effects test for S-KET treated rats run via biased conditioning procedure. Similar to d-AMP, RS-KET significantly produced BS in the development phase and the long-lasting test which result was dependent on CPP procedures. S-KET treatment failed to form BS except that in unbiased group during the development phase. The results of western blot assay reveled 1) drug-related decreases of the GluN1, GluN2A, and GluN2B subunits of NMDA receptor were mainly observed in the nucleus accumbens; 2) the changed levels of three receptor subunits in opposite between ketamine isomers and d-AMP were seen in the dorsal striatum; and 3) region- and drug-dependent changes of dopamine D1 and D2 receptors and dopamine transporter were also detected. Together, differential behavioral effects of ketamine isomers and d-AMP on BS and CPP paradigms could be related to the alternation of glutamate and dopamine neurotransmission systems by drug treatments. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-055 Differential c-Fos expression in rodents with acute heroin exposure *Brianna Brie Evans(1), Sarah Ballard(1), Kyra Newmaster(1), Yongsoo Kim(1), Sue Grigson(1) 1. Department of Neural and Behavioral Sciences, Pennsylvania State College of Medicine, Hershey, PA Keyword: ADDICTION, OPIOID, IMAGING, RODENT While we are familiar with brain areas and pathways that are implicated in opioid use disorder (OUD), we do not have a full understanding of the neural circuits activated upon drug exposure. In order to identify areas of the brain most activated by opioids, we ran a study using transgenic c-Fos-GFP mice and naïve wild-type Sprague-Dawley rats acutely injected with saline interperitoneally (ip) or heroin (10 mg/kg, ip) and examined brain-wide activity patterns using quantitative high-resolution mapping methods. Two-photon tomography was used on the mouse brains and light sheet microscopy on the rat brains. In the mice, we examined c-Fos expression in the Nucleus Accumbens (NAc), Lateral Hypothalamus (LH), Orbitofrontal Cortex (OFC), Cingulate Cortex (CC), Basolateral Amygdala (BLA), Habenula, Nucleus Tractus Solitarus (NTS), Parabrachial Nucleus (PBN), Locus Coeruleus (LC), and Periaqueductal Gray (PAG). These regions are known to be highly activated by drug and most evidenced an increase in c-Fos expression with acute heroin exposure. For the rats, we looked at c-Fos activation patterns in the cortex following the acute administration of saline (ip) or heroin (10 mg/kg, ip), in rats pretreated subcutaneously (sc) with saline or the glucagon-like protein-1 receptor agonist (GLP-1RA), liraglutide. There is some evidence in literature that GLP-1RAs can treat different substance use disorders in rodents and our lab has found that GLP-1RAs can reduce OUD behaviors in rats. Results showed an increase in c-Fos activation in the cortex of the rat following the acute administration of heroin and this activation pattern was reversed by pretreatment with the GLP-1RA, liraglutide. Taken together, the data show the impact of the acute administration of heroin on brain, and reversal of these activation patterns in both mice and rats following treatment with the known satiety agent and GLP-1RA, liraglutide. As such, GLP-1RAs may mitigate not only relapse in drug-experienced subjects, but acquisition of drug-taking as well. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-056 μオピオイド受容体選択的阻害薬の自発運動に対する効果 Effects of a selective μ-opioid receptor antagonist on spontaneous behaviors in mice *北中 順惠(1)、北中 純一(1,2)、田中 康一(2)、五十嵐 健人(3)、富田 和男(3)、西山 信好(2)、佐藤 友昭(3) 1. 兵庫医科大学、2. 兵庫医療大学薬学部、3. 鹿児島大学大学院医歯学総合研究科 *Nobue Kitanaka(1), Junichi Kitanaka(1,2), Koh-ichi Tanaka(2), Kento Igarashi(3), Kazuo Tomita(3), Nobuyoshi Nishiyama(2), Tomoaki Sato(3) 1. Hyogo Col Med, Nishinomiya, Japan, 2. Sch Pharm, Hyogo Univ hlth Sci, Kobe, Japan , 3. Grad Sch Med & Dent Sci, Kagoshima Univ, Kagoshima, Japan Keyword: OPIOID, BEHAVIOR, LOCOMOTION Background: μ-Opioid receptor antagonists have been used for treatment of alcohol dependence as a suppresser for craving. Our research indicated that methamphetamine-induced stereotypy is improved by pretreatment of naloxone. The naloxone action was mimicked by μ- but not κ- or δ-opioid receptor antagonists, indicating that μ-opioid receptor antagonists are likely to suppress brain reward system. In this presentation, we hypothesized that μ-opioid receptor antagonists could inhibit the natural reward as well. Methods: To evaluate natural reward, horizontal running wheels were applied to mice administered with a μ-opioid receptor antagonist β-funaltrexamine (β-FNA). In addition to the rotation of running wheels, horizontal locomotion, amounts of food intake and drinking, and access of food container were measured simultaneously by using a multi-configuration apparatus (0700/1900 light on/off). Male ICR mice were untreated (“naïve”) or treated with a single i.p. injection of 5 mg/kg β-FNA or saline vehicle. The groups of mice were exposed to the multi-configuration apparatus in order to behavioral measures for estimation of the natural reward and the effects of μ-opioid receptor antagonist on the natural reward for three consecutive days (total of 71 h 30 min). Results: In naïve mice, horizontal locomotion increased day by day for three testing days. The increment was parallel to the increase in the rotation of running wheels but not to the number of accesses to food containers. The increase in the rotation of running wheels was sensitized, and the amount of food intake was unchanged in naïve mice. In mice group which was administered with 5 mg/kg β-FNA, the locomotion and thus the rotation of the running wheels decreased especially in the first testing day. The number of accesses to the food containers decreased especially in the first light phase, resulting in the decrease in the total amount of food intake while the number of accesses to the food containers increased in the first dark phases. Discussion: The present results indicate that the use of running wheels is recognized as natural reward in mice. The motivation of use of the running wheels were reduced in mice after treatment with β-FNA, suggesting the suppression of natural reward by μ-opioid receptor antagonists. To confirm this, effects of κ- and δ-opioid receptor antagonists on the rotation of the running wheels are under investigation. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-057 GSK3阻害薬SB216763はモルヒネ作用のうちStraubの挙尾反応を選択的に抑制する SB216763, a glycogen synthase kinase 3 inhibitor, blocks morphine-induced Straub’s tail reaction selectively, but not any other morphine effects *北中 純一(1,2)、北中 順惠(2)、富田 和男(3)、五十嵐 健人(3)、田中 康一(1)、佐藤 友昭(3)、西山 信好(1) 1. 兵庫医療大学薬学部、2. 兵庫医科大学、3. 鹿児島大学大学院医歯学総合研究科 *Junichi Kitanaka(1,2), Nobue Kitanaka(2), Kazuo Tomita(3), Kento Igarashi(3), Koh-ichi Tanaka(1), Tomoaki Sato(3), Nobuyoshi Nishiyama(1) 1. Sch Pharm, Hyogo Univ Hlth Sci, Kobe, Japan, 2. Hyogo Col Med, Nishinomiya, Japan, 3. Grad Sch Med & Dent Sci, Kagoshima Univ, Kagoshima, Japan Keyword: OPIOID, BEHAVIOR, LOCOMOTION Background: Straub’s tail reaction (STR) is a dorsiflexion of the tail that is often vertical to the orientation of the body or curling back over the animal. The phenomenon is thought to be mediated by activation of the opioid receptors because opioid receptor antagonists such as naloxone block the phenomenon. The endpoint of the STR can be identified easier than that of locomotor activity, but behavioral measures of STR can only be achieved by skilled observers. Methods: In this presentation, we investigated the measures of STR by using newly modified, infrared beam sensor-based automated apparatus. The positive tail response was considered as a persistent elevation of the tail at an angle more than 45°. Also, expression levels of GSK3beta and phosphorylated GSK3beta (pGSK3betaser9) were examined in the region of the striatum and nucleus accumbens and in the cerebral cortex after systemic morphine. In constipated mice after morphine, the effect of GSK3 inhibitor on gastrointestinal transit was also examined to reveal whether the action of GSK3 inhibitor such as SB261763 on morphine effects was central and/or peripheral. Results: Mice treated with a single injection of morphine (30 mg/kg) showed a significant STR with a plateau level at a time point of 20 min after morphine challenge. Pretreatment of mice with a selective GSK3 inhibitor SB216763 (0.5 mg/kg) significantly inhibited morphine-induced STR and attenuated the time of STR duration. Horizontal locomotor activity was also attenuated by SB216763. It is of interest to note that mice showing the STR did not exhibit rearing, a search phase of exploratory behavior. In the brain regions tested, expression levels of pGSK3betaser9 in mice after morphine were not altered in mice treated with GSK3 inhibitor in combination with morphine. The pretreatment with GSK3 inhibitor did not improve constipation in morphine-injected mice. Discussion: Although the underlying mechanism is not clear, this data suggests that the inhibitory effect of SB216763 on morphine-induced STR may depend on the blockade of the glycogen synthase kinase-3 signaling pathway. The inhibitory effect of GSK3 inhibitor on morphine-induced STR might not depend on the blockade of GSK3 in terms of phosphorylation levels of pGSK3betaser9. The mechanism of action is likely to be central but not peripheral. Our measurement system is a useful tool for investigating the excitatory effects of morphine in experimental animals. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-058 Sex differences in pathological choice of oxycodone over healthy rewards *Eric Augier(1), Veronika Schwabl(1), Mihai Atudorei(1), Gaëlle Augier(1) 1. Department of Clinical and Experimental Medicine (BKV) Keyword: opioids, sex differences, addiction-like behaviors, choice Use of addictive substances accounts for almost 10% of global disease burden. Opioid misuse in particular has recently become a major concern and a rapidly evolving public health crisis that requires innovative scientific approaches. Oxycodone is one of the most abused prescription opioids, but despite its prevalent misuse, the number of preclinical studies investigating oxycodone-seeking behaviors is relatively limited. Furthermore, opioid use disorder is characterized by a progressive shift of decision making, in which opioids are increasingly chosen over healthy non-drug rewards. Only a subset of people transition from recreational to pathological use. By contrast, in commonly used animal models, nearly all rats learn to self-administer addictive drugs, including opioids and animals have no alternative to drug use. The primary goal of the present study was therefore to investigate potential sex differences in oxycodone self-administration and choice of oxycodone at the expense of healthy rewards. Using operant conditioning, we first trained male and female rats to self-administer oxycodone orally. We found that males earned more oxycodone reinforcers and showed a higher motivation for oxycodone. Blood oxycodone metabolites concentrations were in addition strongly correlated with oxycodone reinforcers earned during self-administration. Finally, we used an exclusive choice-based method to identify both male and female rats that continue to self-administer oxycodone at the expense of a high-value natural reward, a sweet solution, and found that, in agreement with our previous work with alcohol, only a minority of outbred rats (10% or less) choose oxycodone over an alternative high-value reward. Together, our results indicate profound sex differences in oxycodone-related behaviors. They also confirm and extend to oxycodone previous data obtained with preclinical choice models. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-059 報酬の消去に適応するためのアセチルコリン神経細胞の潜在的役割 A potential role of basal forebrain cholinergic neurons in learning of reward omission *向平 妃沙(1)、石野 誠也(1)、鎌田 泰輔(1)、小川 正晃(1) 1. 京都大学大学院医学研究科メディカルイノベーションセンターSKプロジェクト *Hisa Mukohira(1), Seiya Ishino(1), Taisuke Kamada(1), Masaaki Ogawa(1) 1. SK project, Medical Innovation Center, Graduate School of Medicine, Kyoto University Keyword: acetylcholine, basal forebrain, learning, reward How quickly and efficiently animals can adjust their behavior to changing environments is critical for survival. In the face of multiple times of omission of expected reward, animals need to inhibit their behavior to obtain the reward. The neural substrates that modulate the efficiency of learning to inhibit reward-oriented behavior remain unclear. Acetylcholine (ACh) neurons in the basal forebrain (BF) have been implicated in arousal, attention, and learning. Recent studies have shown that ACh neurons in BF respond to reward and punishment in a phasic manner, and enhance cue-reward learning through projection to basolateral amygdala (BLA) (Hangya et al, 2015, Crouse et al, 2020). However, the role of the ACh neurons in inhibitory learning is unknown. We hypothesized that ACh neurons projecting to the hippocampus, which is thought to be critical for hippocampus-dependent memory, might be critical for the inhibitory learning. To address the potential role of the ACh neurons, we employed a behavioral task that we have recently developed (Ishino et al & Ogawa, in preparation). We trained head-restrained rats to push a lever to trigger presentation of a conditioned stimulus and then pull the lever to obtain a probabilistic reward. We recorded ACh transients in the hippocampus and BLA with fiber photometry and activity of ACh neurons in BF with single-cell level calcium imaging. We further examined behavioral effects of optogenetic stimulation of cholinergic axon terminals in hippocampus. We will present data that seem to be consistent with a role in enhancing learning to inhibit reward-oriented behavior in the face of reward omission. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-060 マウスの報酬予測行動における脳内ドーパミン放出動態の解析 Expectation-modulated dynamics of dopamine release during food seeking behavior in mice *柴田 智弘(1)、小澤 貴明(2)、松本 悠真(1)、中村 萌(1)、岩本 涼太郎(1)、尾山 義信(1)、Tom Macpherson(2)、疋田 貴俊(2) 1. 大阪大学理学研究科、2. 大阪大学蛋白質研究所 *Tomohiro Shibata(1), Takaaki Ozawa(2), Yuma Matsumoto(1), Moe Makamura(1), Ryotaro Iwamoto(1), Yoshinobu Oyama(1), Tom Macpherson(2), Takatoshi Hikida(2) 1. Graduate school of science, Osaka university, Japan, 2. Institute for Protein Research, Osaka University, Osaka Japan Keyword: REWARD, DOPAMINE, STRIATUM, IMAGING Positive rewarding outcomes can facilitate the prediction of a future reward and strongly drive goal-directed behavior. On the other hand, negative outcomes suppress current reward seeking behavior and facilitate flexible strategy change. Mesolimbic dopamine neurons, particularly those projecting to the striatum, have been considered as an essential regulatory mechanism underlying flexible appetitive behaviors in animals. Notably, mesolimbic dopamine neurons have been considered to express reward prediction error, as they are strongly activated by unpredicted reward delivery compared predicted reward delivery, and are also strongly inhibited when an expected reward is omitted; however, the precise dynamics of striatal dopamine release during the learning of associations between reward predictive cues and food rewards has not yet been investigated. To address this question, we recorded striatal dopamine release in mice during auditory appetitive conditioning by taking advantage of the fluorescence dopamine sensor GRABDA. GRABDA is engineered by coupling cpGFP to a human dopamine receptor, and fluoresces upon the binding of intrinsic dopamine. In the present study, we trained head-fixed mice in an auditory Pavlovian appetitive conditioning paradigm where one auditory stimulus (CS-High) is followed by a liquid reward delivery (unconditioned stimulus, US) with high probability (80%) but it was delivered with low probability (20%) after the other auditory cue (CS-Low). It was found that dopamine release in the striatum was significantly activated during the CS+ following days of conditioning. Importantly, striatal dopamine release nicely demonstrated typical reward prediction errors, which have been observed in midbrain dopamine neurons. These results suggest the possibility that experience-dependent changes in striatal dopamine release are important for adaptive reward seeking behavior in mice. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-061 恐怖条件づけにおいて表出される条件づけ行動は複数の因子によって変容する Expression patterns of conditioned responses during the fear conditioning are altered by multiple factors. *山本 亮(1)、古山 貴文(1)、今吉 彩菜(1)、伊與部 都世(1)、小野 宗範(1)、加藤 伸郎(1) 1. 金沢医科大学生理学１ *Ryo Yamamoto(1), Takafumi Furuyama(1), Ayana Imayoshi(1), Toyo Iyobe(1), Munenori Ono(1), Nobuo Kato(1) 1. Dept of Physiolosy, Kanazawa Med Univ, Japan Keyword: fear conditioning, conditioned response, freezing, flight In classical fear conditioning, the extent of freezing as a behavioral conditioned response (CR) is commonly used as the index to assess the extent of memory formation. Recently, however, flight behaviors have also been reported as a CR in mice under some situations. In this paper, we examined the possibilities that multiple parameters during the fear conditioning experiment could alter expression patterns of CR in mice. First, we examined the influence of the intensity of the unconditioned stimulus (US) or conditioned stimulus (CS) on expression patterns of CR in mice. We used 2 different intensities for both CS and US in our modified fear conditioning and examined the pattern of CRs. The CS was continuous tones (8 kHz, 20 sec, 90 or 70 dB SPL) followed by the US, which was foot-shocks (1 sec, 0.9 or 0.4 mA). The schedule of fear conditioning was: Day1, CS ×5; Day2, US-CS conditioning ×5; Day3, US-CS conditioning ×5; Day4, CS ×15. The comparison of 4 groups (2-fold US × 2-fold CS) revealed that the intensity of CS (tone loudness) was the primary factor to trigger flight behaviors during CS presentations. Next, we tested whether loud CS presentation is required during conditioning or during the test session, or both, to trigger conditioned flight behaviors. For this purpose, we conditioned the mice with 90 dB CS or 70 dB CS for 2 days and tested the memory by flipping the CS intensity. During the test session, 90 dB CS elicited flight behaviors in the 70 dB conditioned mice, with the opposite loudness combination unsuccessful. This result also supports the idea that CS intensity is the primary factor to trigger conditioned flight behaviors, and the louder CS during the conditioning is not necessary for the expression of the conditioned flight. Finally, we tested the effect of the number of days for conditioning on the expression of conditioned flight behaviors. Surprisingly, when 10 US-CS pairs were delivered on a single day, instead of 5 US-CS pairs on each of 2 consecutive days, no conditioned flight behavior was observed in the test session. This result indicates that repetitive conditionings for several days are required to express conditioned flight behaviors. Consistent with recent reports, our result clearly indicated that freezing is not the sole entity of the CR during the fear conditioning. CR manifestation patterns would be curved by various factors during the US-CS associative learning. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-062 Nucleus accumbens core D1-MSNs microcircuits drive locomotor and reward-related behaviors *Suthinee Attachaipanich(1), Tom Macpherson(1), Takatoshi Hikida(1) 1. Institute for Protein Research, Osaka University Keyword: nucleus accumbens, reward learning, locomotion The nucleus accumbens (NAc) is a critical component of a limbic basal ganglia circuit and is thought to play an important role in reward- and aversion-related learning, including the control of Pavlovian conditioning and goal-directed behavior. Within the NAc, neurons can be anatomically separated into two subpopulations, dopamine D1- and D2-receptor-expressing medium spiny neurons (D1- D2-MSNs). While both NAc D1- and D2-MSNs project to the ventral pallidum (VP), D1-MSNs additionally project to the substantia nigra pars reticulata (SNr). Previous evidence has indicated that NAc D1- and D2-MSN pathways to the VP to control reward and aversive learning, respectively; however, at present, the role of the NAc D1-MSN projection pathway to the SNr is unclear. Given that the SNr is known to contribute to both motor and limbic basal ganglia circuits, in this study we investigated the effect of optogenetic manipulation of the NAc D1-MSN-to-SNr pathway on locomotion and reward-learning tasks. Additionally, we also investigate the optogenetic manipulation of NAc D1-MSN-to-VP pathway in the same behavior tasks for further investigation of the precise behavioral effect underlying specific-output connection from the NAc core. In D1-Cre transgenic mice, we selectively expressed stimulatory channelrhodopsin-2 (ChR2) in NAc D1-MSNs by microinjection of a cre-dependent AAV construct into the NAc core then bilaterally implanted optics fibers above the SNr or the VP. Our findings suggested that optogenetic stimulation of either the D1-MSN-to-SNr or VP pathway increased locomotor activity in an open field test, produced a rewarding effect in self-stimulation and real-time place preference tasks. Interestingly, these projection pathways also enhanced a motivational response in two-choice schedules of reinforcement, especially laser-paired natural reward. Collectively, these results indicate that the D1-MSN-to-SNr projection pathway contributes to locomotion and reward-related learning. However, the activation of NAc core-SNr projection provided weaker impact of locomotion and rewarding effect when compared with VP projection pathway. These suggest the need for further investigation of pathway-specific populations of NAc neurons that will allow for a more precise understanding of the diverse functional roles of the NAc. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-063 長期ストレスによる報酬抑制メカニズムの研究に資する新規ショウジョウバエモデルの確立と分子メカニズムの解明 Chronic stress-induced reward suppression and its molecular mechanism in Drosophila *西塚 悠人(1)、辻 真人(1)、榎本 和生(1,2) 1. 東京大学大学院理学系研究科、2. 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 *Yuto Nishizuka(1), Masato Tsuji(1), Kazuo Emoto(1,2) 1. Graduate School of Science, The University of Tokyo, 2. International Research Center for Neurointelligence (WPI-IRCN), The University of Tokyo Keyword: Chronic stress, reward suppression, serotonin, Drosophila ストレスを一過的に受けた動物は、報酬への応答を抑制すること(以下、報酬抑制)により、危険を回避しエネルギー消費を抑える。一方で、ストレスを長期的に受けた動物は、報酬抑制が過度に持続し、うつ病の中心的症状である無快感症を呈する。では、長期ストレスはどのように報酬抑制をひきおこすのだろうか？この報酬抑制の要因として、長期ストレスによるセロトニン量の減少が着目されてきた。実際、最も広く処方される抗鬱薬はシナプス間隙におけるセロトニン量を増加させることが示唆されている。加えて、マウス脳のセロトニン作動性ニューロン群のうち、報酬系へと投射する集団のみを光遺伝学的手法により活性化すると、報酬への応答が促進されることがわかっている。その一方で、セロトニンは、報酬応答への応答を促進するドーパミンに対して拮抗し、報酬抑制を引き起こすことがわかっている。このように、哺乳類をモデルとした研究からは、セロトニンの報酬抑制に対する寄与については相反する報告がなされている。この１つの原因として、セロトニン作動性ニューロン群は形態・分子・生理学的に多様な集団であり、この多様性が、実験条件の違いによって相反する結果を生み出した可能性が考えられる。 ショウジョウバエはマウスの0.1%程度のニューロン数からなる小さな脳を持ち、遺伝学的に１細胞レベルでニューロン活動を操作できる。そのため、長期ストレスがセロトニンを介して報酬抑制をひきおこすか、その場合どのような神経回路メカニズムによるのか解明できる可能性がある。そこで我々はショウジョウバエにおいて、1. 新規実験系を立ち上げ長期ストレスが報酬抑制をひきおこすか検証し、 2. 長期ストレスによる報酬抑制の原因がセロトニンの減少であるか検証した。ショウジョウバエの報酬として最も一般的なスクロースを用いて、ハエのスクロース応答を計測したところ、長期ストレス負荷によりスクロース応答が低下することが明らかになった。興味深いことに、ヒトに広く処方される抗鬱薬によりストレスによるスクロース応答の抑制が回復した。これらの結果から、長期ストレス負荷によるスクロース応答低下は、ヒトとショウジョウバエにおいて、同様の神経メカニズムを介して惹起される可能性が示唆された。実際、セロトニン合成酵素を欠損したショウジョウバエ変異体は、長期ストレスを負荷せずとも顕著なスクロース応答低下が観察されることから、上記の可能性が支持された。現在、確立したショウジョウバエモデルを用いて、長期ストレス依存的な報酬抑制メカニズムの同定を行っており、特定のセロトニン作動性ニューロンやドーパミン作動性ニューロンの活動変動が関与する可能性を見出している。今後ショウジョウバエモデルを用いて、ストレスが無快感症をひきおこすメカニズムを明らかにしていくとともに、哺乳類との共通メカニズムを理解したい。 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-064 ショウジョウバエ報酬学習におけるドーパミン自己調節機構 Dopamine auto-feedback regulation fine-tunes reward memory in Drosophila *西塔 心路(1)、平松 駿(1)、谷本 拓(1) 1. 東北大学大学院生命科学研究科 *Kokoro Saito(1), Shun Hiramatsu(1), Hiromu Tanimoto(1) 1. Graduate school of life sciences, Tohoku university Keyword: reward memory, dopamine, autoreceptor Dopamine receptors play important roles in a wide range of behaviors, including sensory processing, motor function and reward memory. In Drosophila melanogaster, two dopamine receptors: DopR1 and D2R are localized in axon terminals of the protocerebral anterior medial (PAM) cluster dopaminergic neurons which mediate the reinforcement property of sugar reward in the brain. Based on the sequence similarity, DopR1 is closely related to vertebrate D1 receptors. D2R, as the name indicates, is classified as the fly ortholog to vertebrate D2 receptors. These dopamine receptors in the PAM terminals have a potential to function as autoreceptors that regulate reward signaling and behavioral responses in appetitive learning. Here, we show that DopR1 and D2R in the PAM neurons fine-tune appetitive olfactory memory depending on reward strength. Cell type-specific silencing DopR1 reduced memory performance only at a lower concentration of sugar. On the other hand, the disruption of D2R selectively enhanced memory acquisition at a higher concentration. These results suggest that DopR1 and D2R in the PAM neurons together control the reward gain for learning. This modification of dose-dependent behavioral responses possibly tunes the dynamic range of reward strength. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-065 24時間連続型オペラント箱を用いた学習係数の推定 Estimation of learning parameters using a 24h continous operant box *太田 宏之(1)、佐鳥 玖仁朗(2)、宝田 悠(2)、荒毛 将史(1)、守本 裕司(1)、高橋 達二(2)、石塚 俊晶(1) 1. 防衛医科大学校、2. 東京電機大学 *Hiroyuki Ohta(1), Kuniaki Satori(2), Yu Takarada(2), Masashi Arake(1), Yuji Morimoto(1), Tatsuji Takahashi(2), Toshiaki Ishizuka(1) 1. National Defense Medical College, 2. Tokyo Denki University Keyword: Operant conditioning, Multi armed bandit problem, Reinforcement learning Reinforcement learning models have been used to analyze goal-oriented behaviors of animals and humans. Such models predict future outcomes of selected actions utilizing action values and updating those values in response to the positive and negative outcomes. In most reinforcement learning models, the action values are updated symmetrically based on a common learning rate, that is, in the same way for both positive and negative outcomes. However, animals in environments with scarce rewards may have uneven learning rates. To investigate the asymmetry in learning rates in reward and non-reward, we analyzed the exploration behavior of mice in five armed bandit tasks using a Q-learning model with differential learning rates for positive and negative outcomes. Since operant conditioning in the low reward probability condition requires a long learning period, we developed a 24-hour continuous operant conditioning system with a nest box. By utilizing it, we estimated the positive and negative learning rates. The positive learning rate was significantly higher in a scarce reward environment than in a rich one. Conversely, the negative learning rate was significantly lower in the scarce environment. The positive to negative learning rate ratio was about 10 in the scarce environment and about 2 in the rich environment. We also analyzed the effect of asymmetric learning rates in pseudo simulating environments. The asymmetry causes an overestimation of rare-rewarding events. The overestimation increases total reward acquisition in the scarce environment but disadvantaging impartial exploration. These results suggest that when the reward probability was low, the mice tended to ignore failures and exploit the rare rewards and that this tendency has certain rationality to survive. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-066 内側前頭前皮質から中脳水道周囲灰白質への神経投射の活性化は葛藤環境で恐怖を克服することを促進する Activation of neural projection from the medial prefrontal cortex to the periaqueductal gray facilitates overcoming fear in a conflict context *本宿 雄基(1,2)、曽我 卓司(2)、齋藤 瞭毅(2)、宮上 祐里佳(2)、森 翠(2)、野村 洋(3)、南 雅文(2)、香月 博志(1)、人羅(今村) 菜津子(1,2) 1. 熊本大学大学院生命科学研究部、2. 北海道大学大学院薬学研究院、3. 名古屋市立大学 大学院医学研究科 *HONSHUKU YUKI(1,2), Takuji Soga(2), Ryoki Saito(2), Yurika Miyagami(2), Aoi Mori(2), Hiroshi Nomura(3), Masabumi Minami(2), Hiroshi Katsuki(1), Natsuko Hitora-Imamura(1,2) 1. Grad Sch Pharm, Univ of Kumamoto, Kumamoto, Japan, 2. Grad Sch Pharm, Univ of Hokkaido, Hokkaido, Japan, 3. Grad Sch Med, Nagoya City University , Nagoya, Japan Keyword: selection of behavior, neural circuits, medial prefrontal cortex, periaqueductal gray We are surrounded by various appetitive stimuli which promote behavior and aversive stimuli which prevent behavior. In order to take action in a conflict situation where we face both appetitive and aversive stimuli, we need a willpower: an ability to try to overcome difficulties to achieve goals. Although there are lots of studies focusing on neural activity that promote or prevent behavior, it is not fully understood how these neural activities keep balance in the brain when animals are in a conflict context that has both appetitive and aversive stimuli. We set up a behavioral paradigm in which mice need to explore an experimental context where they might receive electric shocks in order to obtain sucrose solution. Neuronal activity mapping with c-Fos revealed that the medial prefrontal cortex (mPFC) and the periaqueductal gray (PAG) were activated in the conflict context. So, we investigated the role of mPFC-PAG pathway activity in a conflict context. To record the mPFC-PAG activities from freely behaving mice, a genetically encoded calcium indicator, GCaMP6, was expressed selectively in the mPFC-PAG pathway and its fluorescence was detected through an optic fiber. The mPFC-PAG activity was associated with sucrose-seeking behavior. Next, we targeted the mPFC-PAG pathway for optogenetic manipulation by expressing channelrhodopsin-2 or halorhodopsin selectively in the PAG projecting mPFC neurons and phtostimulating the PAG. Optogenetic manipulation of the mPFC-PAG pathway had no effect on reward seeking behavior in the no-conflict context. On the other hand, optogenetic activation and silencing of the mPFC-PAG pathway shortened and prolonged the latency to obtain sucrose, respectively, in the conflict context. In order to examine the effect of mPFC-PAG pathway manipulation on fear memory retrieval, mice were subjected to contextual fear conditioning and examined their freezing with optogenetic manipulation. mPFC-PAG pathway manipulation had no effect on freezing. These results suggest that the mPFC-PAG pathway support overcoming fear to get reward in a conflict environment specific manner. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-067 側坐核に投射するドーパミン作動性ニューロンのGi-DREADDによる化学的阻害はアパシーを誘発するー目標指向性学習と行動の量的低下 Chemogenetic inhibition with Gi-DREADD of dopaminergic neurons projecting to the nucleus accumbens induces apathy-a quantitative reduction of goal-directed leaning and behaviors *王 一妃(1)、高橋 千尋(1)、久恒 辰博(1) 1. 東京大学 *Yifei Wang(1), Chihiro Takahashi(1), Tatsuhiro Hisatsune(1) 1. The University of Tokyo Keyword: dopaminergic neurons, apathy, nucleus accumbens, goal-oriented behaviors Nucleus accumbens (NAc) receives mesolimbic dopaminergic projections from ventral tegmental area (VTA) and glutamatergic projections from prefrontal cortex, amygdala and hippocampus, and sends GABAergic projections to the ventral pallidum. Dopaminergic neurons in the VTA project to the NAc and prefrontal cortex, forming the mesocorticolimbic circuit. One of the key substrates of the reward circuit is NAc and the dopamine in NAc plays an important role in the motivation process. However, the details of such participation are complex and sometimes it may be difficult to sort out. Then regarding reward, aversion, and pleasure sensations, we have to mention one of the clinical non-motor symptoms of Parkinson's disease (PD). That is apathy, which presents early during PD and may even predate onset of motor features. It refers to a set of behavioral, emotional, and cognitive characteristics that include reduced interest and motivation for goal-oriented behavior and affective flattening. Although apathy is a frequent non-motor symptom of PD, the prevalence and pathological basis are not really known and debated. PD patients with apathy present greater motor symptomatology than PD patients without apathy symptoms. Here, to explore the effect of dopaminergic projections from VTA to NAc on apathetic behavior, we employed designer receptors exclusively activated by designer drugs (DREADD) to hyperpolarize neurons through the Gi pathway. We used a combination of retrograde adeno-associated virus and DATIREScre knock-in mice that enables inducing inhibition which is limited in dopaminergic projection to NAc. We demonstrated that, mice with suppressed function of dopaminergic neurons showed a quantitative reduction of goal-directed behaviors: 1. Preference to choose behaviors that putting in little effort to receive little food reward when faced with the choice between low effort- low reward and high effort- high reward in T-maze test. 2. Decrease desire to drink water in the operant drinking test. 3. Reduction of reward-based special learning performance in radial arm maze test. Dopaminergic neural circuits projecting to the NAc are involved in apathy-related behavioral modulation. Gi-DREADD inhibition can trigger a reduction in water or food seeking behavior and prevent goal-directed spatial memory formation. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-068 光遺伝学による背側縫線核セロトニン神経細胞の活性化は正の全脳賦活を誘導する Optogenetic activation of dorsal raphe serotonin neurons induces brain-wide positive responses. *濱田 太陽(1,3)、阿部 欣史(2)、高田 則雄(2)、平良 正和(2)、田中 謙二(2)、銅谷 賢治(1) 1. 沖縄科学技術大学院大学 神経計算ユニット、2. 慶應義塾大学医学部精神神経科学教室、3. （株）アラヤ *Hiro Taiyo Hamada(1,3), Yoshifumi Abe(2), Norio Takata(2), Masakazu Taira(2), Kenji F Tanaka(2), Kenji Doya(1) 1. Neural Computation Unit, Okinawa Institute of Science and Technology, Okinawa, Japan, Tokyo, Japan, 2. Keio University, Tokyo, Japan, 3. Araya Inc., Tokyo, Japan Keyword: serotonin, optogenetic fMRI Serotonin is a neuromodulator controlling multiple functions such as awake-sleep cycle, mood, learning, and decision. Serotonin is a major medication target for major depression, notably by selective serotonin reuptake inhibitors (SSRI). To clarify how the serotonin system controls such functions and medication through its widespread projection and various types of receptors remains unknown, we combined optogenetics and functional MRI using a high S/N 11.7T scanner to measure a brain-wide BOLD response in Tph2-ChR2 transgenic mice. Optogenetic stimulation of dorsal raphe (DR) serotonin neurons, the major serotonergic source, induced extensive activation of reward-related regions including the orbitofrontal cortex (OFC; average peak amplitude of %BOLD: 2.0%, average peak timing: 9.3s), the medial prefrontal cortex (mPFC; 1.9%, 6.0s, the caudate putamen (CPu; 1.3%, 8.0s), and the ventral tegmental area (VTA; 2.7%, 6.9s), the major dopaminergic source, in awake mice (p<0.05, multiple correction). A replication session showed that amplitude and peak timing of %BOLD responses are consistent (amplitude: r=0.886, p<1e-8; timing: r=0.679, p<0.005). These results were contrary to a previous study, which reported widespread deactivation of cortical BOLD signals under anesthesia (Grandjean et al. 2019). We then tested the effect of general anesthesia by isoflurane using the same optogenetic stimulation protocol and found brain-wide negative BOLD responses from OFC, mPFC, the retrosplenial cortex, CPu, and the hippocampal complex (p<0.05, multiple correction). These results indicate that general anesthesia reverses BOLD responses by DR serotonin activation. We further analyzed the BOLD responses by a multiple linear regression model. The BOLD responses in awake state were linearly approximated by a model with the density of DR serotonin projection and gene expression profiles of serotonin receptors including 5HT1A, 5HT1B, 5HT1F, 5HT2A, and 5HT2C subtypes (p<0.05, R2=0.66) with negative weights for inhibitory 5HT1 type receptors and positive weights for excitatory 5HT2 type receptors. BOLD response under general anesthesia was fit with a potentiated weight for inhibitory 5HT1F receptor and a negative weight for excitatory 5HT2c receptor. Our findings accord with the recent discoveries of facilitatory effects of serotonin on reward-guided behaviors and provide a functional anatomical account of brain-wide activation patterns by DR serotonergic stimulation. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-069 ゼブラフィッシュを用いた恐怖応答学習における小脳神経回路の役割の解明 Involvement of cerebellar neural circuits in fear response conditioning in zebrafish *小山 航(1)、細見 椋(1)、松田 光司(2)、川上 浩一(2)、清水 貴史(1,3)、日比 正彦(1,3) 1. 名古屋大学大学院理学研究科、2. 国立遺伝学研究所、3. 名古屋大学生物機能開発利用センター *Wataru Koyama(1), Ryo Hosomi(1), Koji Matsuda(2), Koichi Kawakami(2), Takashi Shimizu(1,3), Masahiko Hibi(1,3) 1. Graduate School of Science, Nagoya University, Nagoya, Japan, 2. National Institute of Genetics, Mishima, Japan, 3. Bioscience and Biotechnology Center, Nagoya University, Nagoya, Japan Keyword: Cerebellum, Fear conditioning, Active avoidance, Zebrafish When animals repeatedly receive a combination of neutral conditional stimulus (CS) and aversive unconditional stimulus (US), they learn the relationship between CS and US, and show conditioned fear responses after CS. They show passive responses such as freezing or panic movements (classical or Pavlovian fear conditioning), or active behavioral responses to avoid aversive stimuli (active avoidance). Previous studies suggested the roles of the cerebellum in classical fear conditioning but it remains elusive whether the cerebellum is involved in active avoidance conditioning. In this study, we analyzed the roles of cerebellar neural circuits during active avoidance in adult zebrafish. When pairs of CS (light) and US (electric shock) were administered to wild-type zebrafish, about half of them displayed active avoidance. The expression of botulinum toxin, which inhibits the release of neurotransmitters, in cerebellar granule cells (GCs) or Purkinje cells (PCs) did not affect conditioning-independent swimming behaviors, but did inhibit active avoidance conditioning. Nitroreductase (NTR)-mediated ablation of PCs with Metronidazole in adult zebrafish also impaired active avoidance. Furthermore, the inhibited transmission of GCs or PCs resulted in reduced fear-conditioned Pavlovian fear responses. Our findings suggest that the zebrafish cerebellum plays an active role in active avoidance conditioning. To understand the details of cerebellar neural circuits in active avoidance and classical fear conditioning, we developed novel optogenetic tools that can control second messengers such as cyclic nucleotides, Ca2+, and diacylglycerol (DAG, an activator of protein kinase C), which have been reported to be involved in cerebellar synaptic plasticity. We evaluated their activity by expressing them in reticulospinal neurons and heart muscle cells in zebrafish larvae. We are currently generating transgenic lines that express the voltage sensors or the optogenetic tools in GCs or PCs by using Gal4-UAS or tetracycline-controlled transactivator (tTA) systems . We shall report our progress of functional imaging and optogenetic analysis of zebrafish cerebellar neural circuits. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-070 恐怖条件づけ中のマウス脳内ドーパミン動態の解析 Dynamics of cortical and subcortical dopamine release during differential auditory fear conditioning in mice. *小澤 貴明(1)、中村 萌(1)、松本 悠真(1)、柴田 智弘(1)、岩本 涼太郎(1)、尾山 賀信(1)、Tom Macpherson(1)、疋田 貴俊(1) 1. 大阪大学蛋白質研究所高次脳機能学研究室 *Takaaki Ozawa(1), Moe Nakamuwa(1), Yuma Matsumoto(1), Shibata Tomohiro(1), Ryotaro Iwamoto(1), Yoshinobu Oyama(1), Tom Macpherson(1), Takatoshi Hikida(1) 1. Institute for Protein Research,Osaka Univ,Osaka,Japan Keyword: dopamine, fear, photometry Prediction and avoidance of future aversive events are vital abilities for survival in animals. Mesolimbic dopamine neurons, particularly those projecting to the striatum, have been considered as an essential regulatory mechanism underlying appetitive behaviors. Notably, dopamine also plays an important role in aversive learnings such as fear conditioning. Previous studies found that striatal dopamine release is inhibited in response to aversive predictive cues and aversive stimuli themselves; however, little is known about how cortical and subcortical dopamine release dynamics change during learning of associations between cues and aversive events. To address this question, we recorded dopamine release in the frontal cortex, the nucleus accumbens and the amygdala in mice during differential auditory fear conditioning by taking advantage of the fluorescence dopamine sensor GRABDA. GRABDA is an engineered intrinsic dopamine sensor made by combining cpGFP with a human dopamine receptor, which increases fluorescence upon the binding of dopamine. In the present study, we trained mice in an auditory fear conditioning paradigm where one auditory stimulus (conditioned stimulus, CS+) is followed by a mild electrical shock (unconditioned stimulus, US) and another is not (CS-). As a result, we found learning dependent changes of dopamine release during CS+. In the nucleus accumbens, dopamine levels were significantly decreased during the CS+ following conditioning. On the other hand, dopamine releases in the frontal cortex and amygdala were increased during CS+ especially after several days of conditioning. These results suggest the possibility that experience-dependent changes in both cortical and subcortical dopamine releases are important for adaptive fear learning and prediction in mice. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-071 オペラント課題中の腹外側線条体ドパミン濃度の計測と報酬予測誤差の推定 Transient decrease of extracellular dopamine concentration in the ventrolateral striatum coincides with negative reward prediction error during food-seeking lever press operant task in mice *井原 慶子(1)、鹿野 悠(1,2)、柳下 祥 (3)、田中 謙二(1)、高田 則雄(1) 1. 慶應義塾大学医学部先端医科学研究所脳科学研究部門、2. スタンフォード大学生物学科、3. 東京大学大学院医学系研究科附属疾患生命工学センター *Keiko Ihara(1), Yu Shikano(1,2), Sho Yagishita(3), Kenji F Tanaka(1), Norio Takata(1) 1. Division of Brain Sciences, Institute for Advanced Medical Research Keio University School of Medicine, Tokyo, Japan, 2. Department of Biology, Stanford University, Stanford, CA, USA, 3. Center for Disease Biology and Integrative Medicine, Faculty of Medicine, The University of Tokyo Keyword: Dopamine, Reward prediction error, GRAB sensor, Methamphetamine How does our reward expectation fluctuate when we continue making efforts to get an uncertain reward or give up? The expectation could be the smallest just before giving up the reward. Alternatively, cumulative efforts might gradually increase reward expectation with anticipation for an approaching reward. Indeed, we observed increase of reward expectation with cumulative lever pressing in an operant conditioning where mice obtained a reward after a constant number of lever-pressing (a fixed ratio [FR] schedule); the larger reward expectation was reflected as larger magnitude of transient decrease (dip) of extracellular dopamine (DA) concentration in the ventral striatum (VS) (Shikano et al. 2021 bioRxiv). We do not know, however, time-course of reward expectation in a progressive ratio (PR) schedule where number of lever presses for obtaining a reward increases exponentially; mice make continuous efforts by repeating a lever-press without recognizing necessary number of lever presses for a reward. To reveal time-course of reward expectation during persistent effort, mice were first trained in a FR task for association of lever pressing and a food pellet reward. Then, we measured extracellular DA dynamics during the PR schedule. Briefly, we injected an adeno associated virus in the VS to express a fluorescent DA sensor protein, GRABDA2m; We used a custom-made fiber photometry system to measure fluorescence fluctuation of the DA sensor. We estimated a magnitude of reward expectation during the PR task by measuring DA dip amplitude when mice checked the food magazine without a reward before completing the required number of a lever press (Shikano et al. 2021 bioRxiv). We found that the DA dip amplitude decreased as the PR task proceeded, and that the dip amplitude was less than 50% of the initial size at the end of the task, indicating that persistent efforts decreased reward expectation. We performed computational modeling to associate mice’s behavioral data with a reward prediction error (RPE). We found that RPE was also decreasing during the PR task, implying that DA dip reflected RPE. We are now analyzing effects of methamphetamine (MAP) on the PR task because MAP alters behavioral activity and motivation of mice. Our computational model might be able to detect such behavioral modulation as　alternation of parameters such as learning rate and inverse temperature in the model. 2022年7月2日　11:00～12:00　沖縄コンベンションセンター 展示棟　ポスター会場1 3P-072 頭部固定下マウスの消去バーストの行動・計算論的分析 Behavioral and computational analysis of extinction burst in head-fixed mice *山田 航太(1)、松井 大(2)、兎田 幸司(1) 1. 慶應義塾大学、2. 北海道大学 *Kota Yamada(1), Hiroshi Matsui(2), Koji Toda(1) 1. Keio University, 2. Hikkaido University Keyword: Operant conditioning, Extinction, Curiosity, Reinforcement learning To behave flexibly in an ever-changing environment, organisms need to gather information about the environment. “Curiosity”, computationally defined based on reward prediction error, novelty, and uncertainty or risk, plays a key role in guiding organisms to explore environments to find unobserved states (Pathak, Agrawal, Efros, and Darrell, 2017). Extinction bursts is the phenomenon that is not explained in traditional learning theory in the sense that the frequency of the learned response is increased after the omission of the predicted outcome. Contrary to the naive intuition that the extinction bursts ordinarily occur after getting an undesirable result in everyday life (such as pressing a keyboard repeatedly or strongly after the character does not show up on the computer display), there have been few empirical studies that support the existence of this phenomenon in experimental settings (Katz and Lattal, 2021). However, recent computational studies of curiosity-driven reinforcement learning and active inference theories suggest that reward prediction errors could evoke preference for the response that produces the errors. Here, we investigate the psychological algorithm behind the extinction bursts by an integrated approach of behavioral analysis, computational modeling, and physiological measurement. First, we built a computational model and conducted simulations to identify the condition to produce the extinction bursts. Second, we experimentally verified whether extinction bursts actually occur in the mice under the conditions identified by the simulations using a head-fixed operant conditioning setup. Third, we measured the pupil size of the mice, and compared the pupil dynamics with variables computed in the computational model. Finally, we confirmed that the extinction burst does not occur in the head-fixed classical conditioning task, suggesting that the extinction burst could be viewed as an active information seeking behavior. Our study revealed that reward prediction errors work as a factor that drives active information seeking.