Updated on 2024/10/04

写真a

 
OKUNO Hiroyuki
 
Organization
Research Field in Medicine and Health Sciences, Medical and Dental Sciences Area Graduate School of Medical and Dental Sciences Advanced Therapeutics Course Functional Biology and Pharmacology Professor
Title
Professor
External link

Degree

  • 博士(医学) ( 2000.3   東京大学 )

  • 修士(理学) ( 1992.3   東京大学 )

Research Areas

  • Life Science / Cognitive and brain science

Education

  • The University of Tokyo

    - 1995.3

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    Country: Japan

  • The University of Tokyo

    1986.4 - 1990.3

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    Country: Japan

Research History

  • Kagoshima University   Research Field in Medicine and Health Sciences, Medical and Dental Sciences Area Graduate School of Medical and Dental Sciences Advanced Therapeutics Course Functional Biology and Pharmacology   Professor

    2018.4

Professional Memberships

  • 日本神経精神薬理学会

    2018.9

  • The Japanese Society of Neuropsychopharmacology

    2018.9

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  • 日本生化学会

    2017.3

  • 日本神経化学会

    2005.1

  • Society for Neuroscience

    1995.4

  • 日本生理学会

    1995.1

  • The Physiological Society of Japan

    1995.1

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  • 日本神経科学学会

    1993.6

  • 日本分子生物学会

    1990.6

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Papers

  • Takakura M., Lam Y.H., Nakagawa R., Ng M.Y., Hu X., Bhargava P., Alia A.G., Gu Y., Wang Z., Ota T., Kimura Y., Morimoto N., Osakada F., Lee A.Y., Leung D., Miyashita T., Du J., Okuno H., Hirano Y. .  Differential second messenger signaling via dopamine neurons bidirectionally regulates memory retention .  Proceedings of the National Academy of Sciences of the United States of America120 ( 36 ) e2304851120   2023.9Reviewed

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    Memory formation and forgetting unnecessary memory must be balanced for adaptive animal behavior. While cyclic AMP (cAMP) signaling via dopamine neurons induces memory formation, here we report that cyclic guanine monophosphate (cGMP) signaling via dopamine neurons launches forgetting of unconsolidated memory in Drosophila. Genetic screening and proteomic analyses showed that neural activation induces the complex formation of a histone H3K9 demethylase, Kdm4B, and a GMP synthetase, Bur, which is necessary and sufficient for forgetting unconsolidated memory. Kdm4B/Bur is activated by phosphorylation through NO-dependent cGMP signaling via dopamine neurons, inducing gene expression, including kek2 encoding a presynaptic protein. Accordingly, Kdm4B/Bur activation induced presynaptic changes. Our data demonstrate a link between cGMP signaling and synapses via gene expression in forgetting, suggesting that the opposing functions of memory are orchestrated by distinct signaling via dopamine neurons, which affects synaptic integrity and thus balances animal behavior.

    DOI: 10.1073/pnas.2304851120

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  • Roberto Pagano, Ahmad Salamian, Janusz Zielinski, Anna Beroun, Maria Nalberczak-Skóra, Edyta Skonieczna, Anna Cały, Nicole Tay, Tobias Banaschewski, Sylvane Desrivières, Antoine Grigis, Hugh Garavan, Andreas Heinz, Rüdiger Brühl, Jean-Luc Martinot, Marie-Laure Paillère Martinot, Eric Artiges, Frauke Nees, Dimitri Papadopoulos Orfanos, Luise Poustka, Sarah Hohmann, Juliane H Fröhner, Michael N Smolka, Nilakshi Vaidya, Henrik Walter, Robert Whelan, Katarzyna Kalita, Haruhiko Bito, Christian P Müller, Gunter Schumann, Hiroyuki Okuno, Kasia Radwanska .  Arc controls alcohol cue relapse by a central amygdala mechanism. .  Molecular psychiatry28 ( 2 ) 733 - 745   2022.11Reviewed International journal

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    Alcohol use disorder (AUD) is a chronic and fatal disease. The main impediment of the AUD therapy is a high probability of relapse to alcohol abuse even after prolonged abstinence. The molecular mechanisms of cue-induced relapse are not well established, despite the fact that they may offer new targets for the treatment of AUD. Using a comprehensive animal model of AUD, virally-mediated and amygdala-targeted genetic manipulations by CRISPR/Cas9 technology and ex vivo electrophysiology, we identify a mechanism that selectively controls cue-induced alcohol relapse and AUD symptom severity. This mechanism is based on activity-regulated cytoskeleton-associated protein (Arc)/ARG3.1-dependent plasticity of the amygdala synapses. In humans, we identified single nucleotide polymorphisms in the ARC gene and their methylation predicting not only amygdala size, but also frequency of alcohol use, even at the onset of regular consumption. Targeting Arc during alcohol cue exposure may thus be a selective new mechanism for relapse prevention.

    DOI: 10.1038/s41380-022-01849-4

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  • David Mahringer, Pawel Zmarz, Hiroyuki Okuno, Haruhiko Bito, Georg B Keller .  Functional correlates of immediate early gene expression in mouse visual cortex. .  Peer community journal2   e45   2022.7Functional correlates of immediate early gene expression in mouse visual cortex.Reviewed International journal

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    During visual development, response properties of layer 2/3 neurons in visual cortex are shaped by experience. Both visual and visuomotor experience are necessary to co-ordinate the integration of bottom-up visual input and top-down motor-related input. Whether visual and visuomotor experience engage different plasticity mechanisms, possibly associated with the two separate input pathways, is still unclear. To begin addressing this, we measured the expression level of three different immediate early genes (IEG) (c-fos, egr1 or Arc) and neuronal activity in layer 2/3 neurons of visual cortex before and after a mouse's first visual exposure in life, and subsequent visuomotor learning. We found that expression levels of all three IEGs correlated positively with neuronal activity, but that first visual and first visuomotor exposure resulted in differential changes in IEG expression patterns. In addition, IEG expression levels differed depending on whether neurons exhibited primarily visually driven or motor-related activity. Neurons with strong motor-related activity preferentially expressed EGR1, while neurons that developed strong visually driven activity preferentially expressed Arc. Our findings are consistent with the interpretation that bottom-up visual input and top-down motor-related input are associated with different IEG expression patterns and hence possibly also with different plasticity pathways.

    DOI: 10.24072/pcjournal.156

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  • Masahiro Sato, Nami Sato-Yamamoto, Ai Wakita, Misako Haraguchi, Manabu Shimonishi, Hiroyuki Okuno .  Direct Injection of Recombinant AAV-Containing Solution into the Oviductal Lumen of Pregnant Mice Caused In Situ Infection of Both Preimplantation Embryos and Oviductal Epithelium. .  International journal of molecular sciences23 ( 9 )   2022.5Reviewed International journal

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    Adeno-associated virus (AAV) vector is an efficient viral-based gene delivery tool used with many types of cells and tissues, including neuronal cells and muscles. AAV serotype 6 (AAV-6), one of numerous AAV serotypes, was recently found to efficiently transduce mouse preimplantation embryos. Furthermore, through coupling with a clustered, regularly interspaced, short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system-a modern genome editing technology-AAV-6 has been shown to effectively create a mutation at a target locus, which relies on isolation of zygotes, in vitro viral infection, and transplantation of the infected embryos to recipient females. Unfortunately, this procedure, termed "ex vivo handling of embryos", requires considerable investment of capital, time, and effort. Direct transduction of preimplantation embryos through the introduction of AAV-6 into the oviductal lumen of pregnant females would be an ideal approach. In this study, we injected various types of recombinant AAV vectors (namely, rAAV-CAG-EGFP-1, -2, -5, and -6, each carrying an enhanced green fluorescent protein [EGFP] cDNA whose expression is under the influence of a cytomegalovirus enhancer + chicken β-actin promoter) into the ampulla region of oviducts in pregnant female mice at Day 0.7 of pregnancy (corresponding to the late 1-cell stage), and EGFP-derived green fluorescence was assessed in the respective morulae. The highest levels of fluorescence were observed in rAAV-CAG-EGFP-6. The oviductal epithelium was distinctly fluorescent. The fluorescence in embryos peaked at the morula stage. Our results indicate that intra-oviductal injection of AAV-6 vectors is the most effective method for transducing zona pellucida-enclosed preimplantation embryos in situ. AAV-6 vectors could be a useful tool in the genetic manipulation of early embryos, as well as oviductal epithelial cells.

    DOI: 10.3390/ijms23094897

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  • Misaki Niu, Atsushi Kasai, Masato Tanuma, Kaoru Seiriki, Hisato Igarashi, Takahiro Kuwaki, Kazuki Nagayasu, Keita Miyaji, Hiroki Ueno, Wataru Tanabe, Kei Seo, Rei Yokoyama, Jin Ohkubo, Yukio Ago, Misuzu Hayashida, Ken-Ichi Inoue, Masahiko Takada, Shun Yamaguchi, Takanobu Nakazawa, Shuji Kaneko, Hiroyuki Okuno, Akihiro Yamanaka, Hitoshi Hashimoto .  Claustrum mediates bidirectional and reversible control of stress-induced anxiety responses. .  Science advances8 ( 11 ) eabi6375   2022.3Reviewed International journal

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    The processing of stress responses involves brain-wide communication among cortical and subcortical regions; however, the underlying mechanisms remain elusive. Here, we show that the claustrum (CLA) is crucial for the control of stress-induced anxiety-related behaviors. A combined approach using brain activation mapping and machine learning showed that the CLA activation serves as a reliable marker of exposure to acute stressors. In TRAP2 mice, which allow activity-dependent genetic labeling, chemogenetic activation of the CLA neuronal ensemble tagged by acute social defeat stress (DS) elicited anxiety-related behaviors, whereas silencing of the CLA ensemble attenuated DS-induced anxiety-related behaviors. Moreover, the CLA received strong input from DS-activated basolateral amygdala neurons, and its circuit-selective optogenetic photostimulation temporarily elicited anxiety-related behaviors. Last, silencing of the CLA ensemble during stress exposure increased resistance to chronic DS. The CLA thus bidirectionally controls stress-induced emotional responses, and its inactivation can serve as a preventative strategy to increase stress resilience.

    DOI: 10.1126/sciadv.abi6375

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  • Mai Takakura, Reiko Nakagawa, Takeshi Ota, Yoko Kimura, Man Yung Ng, Abdalla G Alia, Hiroyuki Okuno, Yukinori Hirano .  Rpd3/CoRest-mediated activity-dependent transcription regulates the flexibility in memory updating in Drosophila. .  Nature communications12 ( 1 ) 628 - 628   2021.12Reviewed International journal

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    Consolidated memory can be preserved or updated depending on the environmental change. Although such conflicting regulation may happen during memory updating, the flexibility of memory updating may have already been determined in the initial memory consolidation process. Here, we explored the gating mechanism for activity-dependent transcription in memory consolidation, which is unexpectedly linked to the later memory updating in Drosophila. Through proteomic analysis, we discovered that the compositional change in the transcriptional repressor, which contains the histone deacetylase Rpd3 and CoRest, acts as the gating mechanism that opens and closes the time window for activity-dependent transcription. Opening the gate through the compositional change in Rpd3/CoRest is required for memory consolidation, but closing the gate through Rpd3/CoRest is significant to limit future memory updating. Our data indicate that the flexibility of memory updating is determined through the initial activity-dependent transcription, providing a mechanism involved in defining memory state.

    DOI: 10.1038/s41467-021-20898-x

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  • Attardo A, Lu J, Kawashima T, Okuno H, Fitzgerald JE, Bito H, Schnitzer MJ .  Long-term consolidation of ensemble neural plasticity patterns in hippocampal area CA1 .  Cell Reports25 ( 3 ) 640 - 650   2018.10Reviewed

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    DOI: 10.1016/j.celrep.2018.09.064

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  • Sami El-Boustani, Jacque P. K. Ip, Vincent Breton-Provencher, Graham W. Knott, Hiroyuki Okuno, Haruhiko Bito, Mriganka Sur .  Locally coordinated synaptic plasticity of visual cortex neurons in vivo .  Science360 ( 6395 ) 1349 - 1354   2018.6Reviewed

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    Plasticity of cortical responses in vivo involves activity-dependent changes at synapses, but the manner in which different forms of synaptic plasticity act together to create functional changes in neurons remains unknown. We found that spike timing–induced receptive field plasticity of visual cortex neurons in mice is anchored by increases in the synaptic strength of identified spines. This is accompanied by a decrease in the strength of adjacent spines on a slower time scale. The locally coordinated potentiation and depression of spines involves prominent AMPA receptor redistribution via targeted expression of the immediate early gene product Arc. Hebbian strengthening of activated synapses and heterosynaptic weakening of adjacent synapses thus cooperatively orchestrate cell-wide plasticity of functional neuronal responses.

    DOI: 10.1126/science.aao0862

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  • Minatohara K, Akiyoshi M, Okuno H .  Role of immediate-early genes in synaptic plasticity and neuronal ensembles underlying the memory trace .  Front Mol Neurosci8   78   2016.1Role of immediate-early genes in synaptic plasticity and neuronal ensembles underlying the memory traceReviewed International journal

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  • Kawashima T, Okuno H, Bito H. .  A new era for functional labeling of neurons: activity-dependent promoters have come of age .  Front Neural Circuits8   37   2014.4A new era for functional labeling of neurons: activity-dependent promoters have come of ageReviewed International journal

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  • Okuno H, Akashi K, Ishii Y, Yagishita-Kyo N, Suzuki K, Nonaka M, Kawashima T, Fujii H, Takemoto-Kimura S, Abe M, Natsume R, Chowdhury S, Sakimura K, Worley PF, Bito H. .  Inverse synaptic tagging of inactive synapses via dynamic interaction of Arc/Arg3.1 with CaMKIIβ. .  Cell149 ( 4 ) 886 - 898   2012.5Inverse synaptic tagging of inactive synapses via dynamic interaction of Arc/Arg3.1 with CaMKIIβ.Reviewed International journal

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    DOI: 10.1016/j.cell.2012.02.062

  • Tse D, Takeuchi T, Kakeyama M, Kajii Y, Okuno H, Tohyama C, Bito H, Morris RG. .  Schema-dependent gene activation and memory encoding in neocortex. .  Science333 ( 6044 ) 891 - 895   2011.8Schema-dependent gene activation and memory encoding in neocortex.

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  • Okuno H. .  Regulation and function of immediate-early genes in the brain: beyond neuronal activity markers. .  Neuroscience Research69 ( 3 ) 175 - 186   2010.12Regulation and function of immediate-early genes in the brain: beyond neuronal activity markers.

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    DOI: 10.1016/j.neures.2010.12.007

  • Redondo RL, Okuno H, Spooner PA, Frenguelli BG, Bito H, Morris RG. .  Synaptic tagging and capture: differential role of distinct calcium/calmodulin kinases in protein synthesis-dependent long-term potentiation. .  J Neurosci30 ( 14 ) 4981 - 4989   2010.4Synaptic tagging and capture: differential role of distinct calcium/calmodulin kinases in protein synthesis-dependent long-term potentiation.

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  • Kawashima T, Okuno H, Nonaka M, Adachi-Morishima A, Kyo N, Okamura M, Takemoto-Kimura S, Worley PF, Bito H. .  Synaptic activity-responsive element in the Arc/Arg3.1 promoter essential for synapse-to-nucleus signaling in activated neurons. .  Proc Natl Acad Sci U S A.106 ( 1 ) 316 - 321   2009.1Synaptic activity-responsive element in the Arc/Arg3.1 promoter essential for synapse-to-nucleus signaling in activated neurons.

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    DOI: 10.1073/pnas.0806518106

  • Chowdhury S, Shepherd JD, Okuno H, Lyford G, Petralia RS, Plath N, Kuhl D, Huganir RL, Worley PF. .  Arc/Arg3.1 interacts with the endocytic machinery to regulate AMPA receptor trafficking. .  Neuron52 ( 3 ) 445 - 459   2006.11Arc/Arg3.1 interacts with the endocytic machinery to regulate AMPA receptor trafficking.

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  • Li YX, Hashimoto T, Tokuyama W, Miyashita Y, Okuno H. .  Spatiotemporal dynamics of brain-derived neurotrophic factor mRNA induction in the vestibulo-olivary network during vestibular compensation. .  J Neurosci.21 ( 8 ) 2738 - 2738   2001.4Spatiotemporal dynamics of brain-derived neurotrophic factor mRNA induction in the vestibulo-olivary network during vestibular compensation.

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    DOI: 10.1523/JNEUROSCI.21-08-02738.2001

  • Tokuyama W, Okuno H, Hashimoto T, Xin Li Y, Miyashita Y. .  BDNF upregulation during declarative memory formation in monkey inferior temporal cortex. .  Nat Neurosci.3 ( 11 ) 1134 - 1142   2000.11BDNF upregulation during declarative memory formation in monkey inferior temporal cortex.Reviewed International journal

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  • Okuno H, Tokuyama W, Li YX, Hashimoto T, Miyashita Y. .  Quantitative evaluation of neurotrophin and trk mRNA expression in visual and limbic areas along the occipito-temporo-hippocampal pathway in adult macaque monkeys. .  J Comp Neurol.408 ( 3 ) 378 - 398   1999.6Quantitative evaluation of neurotrophin and trk mRNA expression in visual and limbic areas along the occipito-temporo-hippocampal pathway in adult macaque monkeys.

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  • Okuno H, Miyashita Y. .  Expression of the transcription factor Zif268 in the temporal cortex of monkeys during visual paired associate learning. .  Eur J Neurosci.8 ( 10 ) 2118 - 2128   1996.10Expression of the transcription factor Zif268 in the temporal cortex of monkeys during visual paired associate learning.Reviewed International journal

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    DOI: 10.1111/j.1460-9568.1996.tb00733.x

  • Okuno H, Akahori A, Sato H, Xanthoudakis S, Curran T, Iba H. .  Escape from redox regulation enhances the transforming activity of Fos. .  Oncogene8 ( 3 ) 695 - 701   1993.3Escape from redox regulation enhances the transforming activity of Fos.Reviewed International journal

  • Itoh M, Okuno H, Yamada D, Yamashita M, Abe M, Natsume R, Kaizuka T, Sakimura K, Hoshino M, Mishina M, Wada K, Sekiguchi M, Hayashi T. .  Perturbed expression pattern of the immediate early gene Arc in the dentate gyrus of GluA1 C-terminal palmitoylation-deficient mice .  Neuropsychopharmacol Reports39 ( 1 ) 61 - 66   2109.3Reviewed

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    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Neuropsychopharmacology Reports  

    DOI: 10.1002/npr2.12044

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  • Kaneko T., Oura A., Imai Y., Kusumoto-Yoshida I., Kanekura T., Okuno H., Kuwaki T., Kashiwadani H. .  Orexin neurons play contrasting roles in itch and pain neural processing via projecting to the periaqueductal gray .  Communications Biology7 ( 1 ) 290   2024.12Reviewed

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    Authorship:Lead author, Last author, Corresponding author   Language:Japanese   Publisher:Communications Biology  

    Pain and itch are recognized as antagonistically regulated sensations; pain suppresses itch, whilst pain inhibition enhances itch. The neural mechanisms at the central nervous system (CNS) underlying these pain-itch interactions still need to be explored. Here, we revealed the contrasting role of orexin-producing neurons (ORX neurons) in the lateral hypothalamus (LH), which suppresses pain while enhancing itch neural processing, by applying optogenetics to the acute pruritus and pain model. We also revealed that the circuit of ORX neurons from LH to periaqueductal gray regions served in the contrasting modulation of itch and pain processing using optogenetic terminal inhibition techniques. Additionally, by using an atopic dermatitis model, we confirmed the involvement of ORX neurons in regulating chronic itch processing, which could lead to a novel therapeutic target for persistent pruritus in clinical settings. Our findings provide new insight into the mechanism of antagonistic regulation between pain and itch in the CNS.

    DOI: 10.1038/s42003-024-05997-x

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  • Roberto Pagano, Ahmad Salamian, Janusz Zielinski, Anna Beroun, Maria Nalberczak-Skóra, Edyta Skonieczna, Anna Cały, Nicole Tay, Tobias Banaschewski, Sylvane Desrivières, Antoine Grigis, Hugh Garavan, Andreas Heinz, Rüdiger Brühl, Jean-Luc Martinot, Marie-Laure Paillère Martinot, Eric Artiges, Frauke Nees, Dimitri Papadopoulos Orfanos, Luise Poustka, Sarah Hohmann, Juliane H Fröhner, Michael N Smolka, Nilakshi Vaidya, Henrik Walter, Robert Whelan, Katarzyna Kalita, Haruhiko Bito, Christian P Müller, Gunter Schumann, Hiroyuki Okuno, Kasia Radwanska .  Correction: Arc controls alcohol cue relapse by a central amygdala mechanism. .  Molecular psychiatry   2022.12Reviewed International journal

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  • 城山 優治, 後藤 史子, 小川 糸音, 吉田 進昭, 尾藤 晴彦, 真鍋 俊也, 奥野 浩行 .  Gastrin-releasing peptideは急性ストレス下で、amygdalostriatal transition areaを通じて恐怖学習を調節する .  日本生化学会大会プログラム・講演要旨集95回   1P - 353   2022.11Gastrin-releasing peptideは急性ストレス下で、amygdalostriatal transition areaを通じて恐怖学習を調節するReviewed

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  • Shiina Nobuyuki, Okuno Hiroyuki .  Expanding evidence of fundamental regulation for biological processes by liquid–liquid phase separation .    94 ( 4 ) 483 - 484   2022.8Reviewed

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    DOI: 10.14952/seikagaku.2022.940483

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  • Akiko Oota-Ishigaki, Keizo Takao, Daisuke Yamada, Masayuki Sekiguchi, Masayuki Itoh, Yumie Koshidata, Manabu Abe, Rie Natsume, Masaki Kaneko, Toma Adachi, Toshie Kaizuka, Nami Suzuki, Kenji Sakimura, Hiroyuki Okuno, Keiji Wada, Masayoshi Mishina, Tsuyoshi Miyakawa, Takashi Hayashi .  Prolonged contextual fear memory in AMPA receptor palmitoylation-deficient mice. .  Neuropsychopharmacology : official publication of the American College of Neuropsychopharmacology47 ( 12 ) 2150 - 2159   2022.5Reviewed International journal

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    Long-lasting fear-related disorders depend on the excessive retention of traumatic fear memory. We previously showed that the palmitoylation-dependent removal of synaptic α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors prevents hyperexcitation-based epileptic seizures and that AMPA receptor palmitoylation maintains neural network stability. In this study, AMPA receptor subunit GluA1 C-terminal palmitoylation-deficient (GluA1C811S) mice were subjected to comprehensive behavioral battery tests to further examine whether the mutation causes other neuropsychiatric disease-like symptoms. The behavioral analyses revealed that palmitoylation-deficiency in GluA1 is responsible for characteristic prolonged contextual fear memory formation, whereas GluA1C811S mice showed no impairment of anxiety-like behaviors at the basal state. In addition, fear generalization gradually increased in these mutant mice without affecting their cued fear. Furthermore, fear extinction training by repeated exposure of mice to conditioned stimuli had little effect on GluA1C811S mice, which is in line with augmentation of synaptic transmission in pyramidal neurons in the basolateral amygdala. In contrast, locomotion, sociability, depression-related behaviors, and spatial learning and memory were unaffected by the GluA1 non-palmitoylation mutation. These results indicate that impairment of AMPA receptor palmitoylation specifically causes posttraumatic stress disorder (PTSD)-like symptoms.

    DOI: 10.1038/s41386-022-01347-9

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  • Fumiko Goto, Yuji Kiyama, Itone Ogawa, Hiroyuki Okuno, Taeko Ichise, Hirotake Ichise, Motonobu Anai, Tatsuhiko Kodama, Nobuaki Yoshida, Haruhiko Bito, Toshiya Manabe .  Gastrin-releasing peptide regulates fear learning under stressed conditions via activation of the amygdalostriatal transition area .  Molecular Psychiatry27 ( 3 ) 1694 - 1703   2022Reviewed

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    Authorship:Lead author, Last author, Corresponding author   Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    DOI: 10.1038/s41380-021-01408-3

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    Other Link: https://www.nature.com/articles/s41380-021-01408-3

  • Tetsuo Ohnishi, Yuji Kiyama, Fumiko Arima-Yoshida, Mitsutaka Kadota, Tomoe Ichikawa, Kazuyuki Yamada, Akiko Watanabe, Hisako Ohba, Kaori Tanaka, Akihiro Nakaya, Yasue Horiuchi, Yoshimi Iwayama, Manabu Toyoshima, Itone Ogawa, Chie Shimamoto-Mitsuyama, Motoko Maekawa, Shabeesh Balan, Makoto Arai, Mitsuhiro Miyashita, Kazuya Toriumi, Yayoi Nozaki, Rumi Kurokawa, Kazuhiro Suzuki, Akane Yoshikawa, Tomoko Toyota, Toshihiko Hosoya, Hiroyuki Okuno, Haruhiko Bito, Masanari Itokawa, Shigehiro Kuraku, Toshiya Manabe, Takeo Yoshikawa .  Cooperation of LIM domain-binding 2 (LDB2) with EGR in the pathogenesis of schizophrenia. .  EMBO molecular medicine13 ( 4 ) e12574   2021.4Reviewed International journal

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    Genomic defects with large effect size can help elucidate unknown pathologic architecture of mental disorders. We previously reported on a patient with schizophrenia and a balanced translocation between chromosomes 4 and 13 and found that the breakpoint within chromosome 4 is located near the LDB2 gene. We show here that Ldb2 knockout (KO) mice displayed multiple deficits relevant to mental disorders. In particular, Ldb2 KO mice exhibited deficits in the fear-conditioning paradigm. Analysis of the amygdala suggested that dysregulation of synaptic activities controlled by the immediate early gene Arc is involved in the phenotypes. We show that LDB2 forms protein complexes with known transcription factors. Consistently, ChIP-seq analyses indicated that LDB2 binds to > 10,000 genomic sites in human neurospheres. We found that many of those sites, including the promoter region of ARC, are occupied by EGR transcription factors. Our previous study showed an association of the EGR family genes with schizophrenia. Collectively, the findings suggest that dysregulation in the gene expression controlled by the LDB2-EGR axis underlies a pathogenesis of subset of mental disorders.

    DOI: 10.15252/emmm.202012574

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  • Hirotaka Fujita, Ryota Oikawa, Mayu Hayakawa, Fumiaki Tomoike, Yasuaki Kimura, Hiroyuki Okuno, Yoshiki Hatashita, Carolina Fiallos Oliveros, Haruhiko Bito, Toshio Ohshima, Satoshi Tsuneda, Hiroshi Abe, Takafumi Inoue .  Quantification of native mRNA dynamics in living neurons using fluorescence correlation spectroscopy and reduction-triggered fluorescent probes. .  The Journal of biological chemistry295 ( 23 ) 7923 - 7940   2020.4Reviewed International journal

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    RNA localization in subcellular compartments is essential for spatial and temporal regulation of protein expression in neurons. Several techniques have been developed to visualize mRNAs inside cells, but the study of the behavior of endogenous and nonengineered mRNAs in living neurons has just started. In this study, we combined reduction-triggered fluorescent (RETF) probes and fluorescence correlation spectroscopy (FCS) to investigate the diffusion properties of activity-regulated cytoskeleton-associated protein (Arc) and inositol 1,4,5-trisphosphate receptor type 1 (Ip3r1) mRNAs. This approach enabled us to discriminate between RNA-bound and unbound fluorescent probes and to quantify mRNA diffusion parameters and concentrations in living rat primary hippocampal neurons. Specifically, we detected the induction of Arc mRNA production after neuronal activation in real time. Results from computer simulations with mRNA diffusion coefficients obtained in these analyses supported the idea that free diffusion is incapable of transporting mRNA of sizes close to those of Arc or Ip3r1 to distal dendrites. In conclusion, the combined RETF-FCS approach reported here enables analyses of the dynamics of endogenous, unmodified mRNAs in living neurons, affording a glimpse into the intracellular dynamics of RNA in live cells.

    DOI: 10.1074/JBC.RA119.010921

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  • Koshi Murata, Tomoki Kinoshita, Yugo Fukazawa, Kenta Kobayashi, Kazuto Kobayashi, Kazunari Miyamichi, Hiroyuki Okuno, Haruhiko Bito, Yoshio Sakurai, Masahiro Yamaguchi, Kensaku Mori, Hiroyuki Manabe .  GABAergic neurons in the olfactory cortex projecting to the lateral hypothalamus in mice. .  Scientific reports9 ( 1 ) 7132 - 7132   2019.12Reviewed International journal

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    Olfaction guides goal-directed behaviours including feeding. To investigate how central olfactory neural circuits control feeding behaviour in mice, we performed retrograde tracing from the lateral hypothalamus (LH), an important feeding centre. We observed a cluster of retrogradely labelled cells distributed in the posteroventral region of the olfactory peduncle. Histochemical analyses revealed that the majority of these retrogradely labelled projection neurons expressed glutamic acid decarboxylase 65/67 (GAD65/67), but not vesicular glutamate transporter 1 (VGluT1). We named this region containing GABAergic projection neurons the ventral olfactory nucleus (VON) to differentiate it from the conventional olfactory peduncle. VON neurons were less immunoreactive for DARPP-32, a striatal neuron marker, compared to neurons in the olfactory tubercle and nucleus accumbens, which distinguished the VON from the ventral striatum. Fluorescent labelling confirmed putative synaptic contacts between VON neurons and olfactory bulb projection neurons. Rabies-virus-mediated trans-synaptic labelling revealed that VON neurons received synaptic inputs from the olfactory bulb, other olfactory cortices, horizontal limb of the diagonal band, and prefrontal cortex. Collectively, these results identify novel GABAergic projection neurons in the olfactory cortex that may integrate olfactory sensory and top-down inputs and send inhibitory output to the LH, which may modulate odour-guided LH-related behaviours.

    DOI: 10.1038/s41598-019-43580-1

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  • Suzuki A.Z. .  A clickable caging group as a new platform for modular caged compounds with improved photochemical properties .  Chemical Communications55 ( 4 ) 451 - 454   2019

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    DOI: 10.1039/c8cc07981a

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  • Masayuki Itoh, Mariko Yamashita, Masaki Kaneko, Hiroyuki Okuno, Manabu Abe, Maya Yamazaki, Rie Natsume, Daisuke Yamada, Toshie Kaizuka, Reiko Suwa, Kenji Sakimura, Masayuki Sekiguchi, Keiji Wada, Mikio Hoshino, Masayoshi Mishina, Takashi Hayashi .  Deficiency of AMPAR-Palmitoylation Aggravates Seizure Susceptibility. .  The Journal of neuroscience : the official journal of the Society for Neuroscience38 ( 47 ) 10220 - 10235   2018.11Reviewed

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    Synaptic AMPAR expression controls the strength of excitatory synaptic transmission and plasticity. An excess of synaptic AMPARs leads to epilepsy in response to seizure-inducible stimulation. The appropriate regulation of AMPARs plays a crucial role in the maintenance of the excitatory/inhibitory synaptic balance; however, the detailed mechanisms underlying epilepsy remain unclear. Our previous studies have revealed that a key modification of AMPAR trafficking to and from postsynaptic membranes is the reversible, posttranslational S-palmitoylation at the C-termini of receptors. To clarify the role of palmitoylation-dependent regulation of AMPARs in vivo, we generated GluA1 palmitoylation-deficient (Cys811 to Ser substitution) knock-in mice. These mutant male mice showed elevated seizure susceptibility and seizure-induced neuronal activity without impairments in synaptic transmission, gross brain structure, or behavior at the basal level. Disruption of the palmitoylation site was accompanied by upregulated GluA1 phosphorylation at Ser831, but not at Ser845, in the hippocampus and increased GluA1 protein expression in the cortex. Furthermore, GluA1 palmitoylation suppressed excessive spine enlargement above a certain size after LTP. Our findings indicate that an abnormality in GluA1 palmitoylation can lead to hyperexcitability in the cerebrum, which negatively affects the maintenance of network stability, resulting in epileptic seizures.SIGNIFICANCE STATEMENT AMPARs predominantly mediate excitatory synaptic transmission. AMPARs are regulated in a posttranslational, palmitoylation-dependent manner in excitatory synapses of the mammalian brain. Reversible palmitoylation dynamically controls synaptic expression and intracellular trafficking of the receptors. Here, we generated GluA1 palmitoylation-deficient knock-in mice to clarify the role of AMPAR palmitoylation in vivo We showed that an abnormality in GluA1 palmitoylation led to hyperexcitability, resulting in epileptic seizure. This is the first identification of a specific palmitoylated protein critical for the seizure-suppressing process. Our data also provide insight into how predicted receptors such as AMPARs can effectively preserve network stability in the brain. Furthermore, these findings help to define novel key targets for developing anti-epileptic drugs.

    DOI: 10.1523/JNEUROSCI.1590-18.2018

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  • Keietsu Kikuchi, Daisuke Ihara, Mamoru Fukuchi, Hiroki Tanabe, Yuta Ishibashi, Junya Tsujii, Masaaki Tsuda, Marisa Kaneda, Hiroyuki Sakagami, Hiroyuki Okuno, Haruhiko Bito, Yuya Yamazaki, Mitsuru Ishikawa, Akiko Tabuchi .  Involvement of SRF coactivator MKL2 in BDNF-mediated activation of the synaptic activity-responsive element in the Arc gene. .  Journal of neurochemistry148 ( 2 ) 204 - 218   2018.11Reviewed

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    The expression of immediate early genes (IEGs) is thought to be an essential molecular basis of neuronal plasticity for higher brain function. Many IEGs contain serum response element in their transcriptional regulatory regions and their expression is controlled by serum response factor (SRF). SRF is known to play a role in concert with transcriptional cofactors. However, little is known about how SRF cofactors regulate IEG expression during the process of neuronal plasticity. We hypothesized that one of the SRF-regulated neuronal IEGs, activity-regulated cytoskeleton-associated protein (Arc; also termed Arg3.1), is regulated by an SRF coactivator, megakaryoblastic leukemia (MKL). To test this hypothesis, we initially investigated which binding site of the transcription factor or SRF cofactor contributes to brain-derived neurotrophic factor (BDNF)-induced Arc gene transcription in cultured cortical neurons using transfection and reporter assays. We found that BDNF caused robust induction of Arc gene transcription through a cAMP response element, binding site of myocyte enhancer factor 2, and binding site of SRF in an Arc enhancer, the synaptic activity-responsive element (SARE). Regardless of the requirement for the SRF-binding site, the binding site of a ternary complex factor, another SRF cofactor, did not affect BDNF-mediated Arc gene transcription. In contrast, chromatin immunoprecipitation revealed occupation of MKL at the SARE. Furthermore, knockdown of MKL2, but not MKL1, significantly decreased BDNF-mediated activation of the SARE. Taken together, these findings suggest a novel mechanism by which MKL2 controls the Arc SARE in response to BDNF stimulation.

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  • Chawla MK, Gray DT, Nguyen C, Dhaliwal H, Zempare M, Okuno H, Huentelman MJ, Barnes CA. .  Seizure-induced Arc mRNA expression thresholds in rat hippocampus and perirhinal cortex .  Frontiers in Systems Neuroscience12   53   2018.11Seizure-induced Arc mRNA expression thresholds in rat hippocampus and perirhinal cortexReviewed

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    DOI: 10.3389/fnsys.2018.00053

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  • Yamada M, Suzuki Y, Nagasaki SC, Okuno H, Imayoshi I .  Light Control of the Tet Gene Expression System in Mammalian Cells. .  Cell reports25 ( 2 ) 487 - 500   2018.10Reviewed

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    DOI: 10.1016/j.celrep.2018.09.026

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  • Nishiguchi KM, Fujita K, Tokashiki N, Komamura H, Takemoto-Kimura S, Okuno H, Bito H, Nakazawa T .  Retained Plasticity and Substantial Recovery of Rod-Mediated Visual Acuity at the Visual Cortex in Blind Adult Mice with Retinal Dystrophy. .  Molecular therapy : the journal of the American Society of Gene Therapy26 ( 10 ) 2397 - 2406   2018.10Retained Plasticity and Substantial Recovery of Rod-Mediated Visual Acuity at the Visual Cortex in Blind Adult Mice with Retinal Dystrophy.Reviewed

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    DOI: 10.1016/j.ymthe.2018.07.012

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  • Hiroyuki Okuno, Keiichiro Minatohara, Haruhiko Bito .  Inverse synaptic tagging: An inactive synapse-specific mechanism to capture activity-induced Arc/arg3.1 and to locally regulate spatial distribution of synaptic weights. .  Seminars in cell & developmental biology77   43 - 50   2018.5Inverse synaptic tagging: An inactive synapse-specific mechanism to capture activity-induced Arc/arg3.1 and to locally regulate spatial distribution of synaptic weights.Invited Reviewed

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    Long-lasting forms of synaptic plasticity such as long-term potentiation (LTP) and long-term depression (LTD) are fundamental cellular mechanisms underlying learning and memory. The synaptic tagging and capture (STC) hypothesis has provided a theoretical framework on how products of activity-dependent genes may interact with potentiated synapses to facilitate and maintain such long-lasting synaptic plasticity. Although Arc/arg3.1 was initially assumed to participate in STC processes during LTP, accumulating evidence indicated that Arc/arg3.1 might rather contribute in weakening of synaptic weights than in their strengthening. In particular, analyses of Arc/Arg3.1 protein dynamics and function in the dendrites after plasticity-inducing stimuli have revealed a new type of inactivity-dependent redistribution of synaptic weights, termed "inverse synaptic tagging". The original synaptic tagging and inverse synaptic tagging likely co-exist and are mutually non-exclusive mechanisms, which together may help orchestrate the redistribution of synaptic weights and promote the enhancement and maintenance of their contrast between potentiated and non-potentiated synapses during the late phase of long-term synaptic plasticity. In this review, we describe the inverse synaptic tagging mechanism that controls synaptic dynamics of Arc/Arg3.1, an immediate early gene product which is captured and preferentially targeted to non-potentiated synapses, and discuss its impact on neuronal circuit refinement and cognitive function.

    DOI: 10.1016/j.semcdb.2017.09.025

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  • Oswald Steward, Kelly Matsudaira Yee, Shannon Farris, Patricia S. Pirbhoy, Paul Worley, Kohji Okamura, Hiroyuki Okuno, Haruhiko Bito .  Delayed degradation and impaired dendritic delivery of intron-lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc transgenic mice .  Frontiers in Molecular Neuroscience10   435 - 435   2018.1Delayed degradation and impaired dendritic delivery of intron-lacking EGFP-Arc/Arg3.1 mRNA in EGFP-Arc transgenic miceReviewed

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    Arc is a unique immediate early gene (IEG) whose expression is induced as synapses are modified during learning. Newly-synthesized Arc mRNAis rapidly transported throughout dendrites and localizes near recently activated synapses. Arc mRNA levels are regulated by rapid degradation, which is accelerated by synaptic activity in a translation-dependent process. One possible mechanism is nonsense-mediated mRNA decay (NMD), which depends on the presence of a splice junction in the 3′ UTR. Here, we test this hypothesis using transgenic mice that express EGFP-Arc. Because the transgene was constructed fromArc cDNA, it lacks intron structures in the 3′ UTR that are present in the endogenous Arc gene. NMD depends on the presence of proteins of the exon junction complex (EJC) downstream of a stop codon, so EGFP-Arc mRNA should not undergo NMD. Assessment of Arc mRNA rundown in the presence of the transcription inhibitor actinomycin-D confirmed delayed degradation of EGFP-Arc mRNA. EGFP-Arc mRNA and protein are expressed at much higher levels in transgenic mice under basal and activated conditions but EGFP-Arc mRNA does not enter dendrites efficiently. In a physiological assay in which cycloheximide (CHX) was infused after induction of Arc by seizures, there were increases in endogenous Arc mRNA levels consistent with translation-dependent Arc mRNA decay but this was not seen with EGFP-Arc mRNA. Taken together, our results indicate: (1) Arc mRNA degradation occurs via a mechanism with characteristics of NMD
    (2) rapid dendritic delivery of newly synthesized Arc mRNA after induction may depend in part on prior splicing of the 3′ UTR.

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  • Sakiko Honjoh, Luisa de Vivo, Hiroyuki Okuno, Haruhiko Bito, Giulio Tononi, Chiara Cirelli .  Higher Arc Nucleus-to-Cytoplasm Ratio during Sleep in the Superficial Layers of the Mouse Cortex .  FRONTIERS IN NEURAL CIRCUITS11   60 - 60   2017.8Higher Arc Nucleus-to-Cytoplasm Ratio during Sleep in the Superficial Layers of the Mouse CortexReviewed

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    The activity-regulated cytoskeleton associated protein Arc is strongly and quickly upregulated by neuronal activity, synaptic potentiation and learning. Arc entry in the synapse is followed by the endocytosis of glutamatergic AMPA receptors (AMPARs), and its nuclear accumulation has been shown in vitro to result in a small decline in the transcription of the GluA1 subunit of AMPARs. Since these effects result in a decline in synaptic strength, we asked whether a change in Arc dynamics may temporally correlate with sleep-dependent GluA1 down-regulation. We measured the ratio of nuclear to cytoplasmic Arc expression (Arc Nuc/Cyto) in the cerebral cortex of EGFP-Arc transgenic mice that were awake most of the night and then perfused immediately before lights on (W mice), or were awake most of the night and then allowed to sleep (S mice) or sleep deprived (SD mice) for the first 2 h of the light phase. In primary motor cortex (M1), neurons with high levels of nuclear Arc (High Arc cells) were present in all mice, but in these cells Arc Nuc/Cyto was higher in S mice than in W mice and, importantly, similar to 15% higher in S mice than in SD mice collected at the same time of day, ruling out circadian effects. Greater Arc Nuc/Cyto with sleep was observed in the superficial layers of M1, but not in the deep layers. In High Arc cells, Arc Nuc/Cyto was also similar to 15%-30% higher in S mice than in W and SD mice in the superficial layers of primary somatosensory cortex (S1) and cingulate cortex area 1 (Cg1). In High Arc Cells of Cg1, Arc Nuc/Cyto and cytoplasmic levels of GluA1 immunoreactivities in the soma were also negatively correlated, independent of behavioral state. Thus, Arc moves to the nucleus during both sleep and wake, but its nuclear to cytoplasmic ratio increases with sleep in the superficial layers of several cortical areas. It remains to be determined whether the relative increase in nuclear Arc contributes significantly to the overall decline in the strength of excitatory synapses that occurs during sleep. Similarly, it remains to be determined whether the entry of Arc into specific synapses is gated by sleep.

    DOI: 10.3389/fncir.2017.00060

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  • Jenks KR, Kim T, Pastuzyn ED, Okuno H, Taibia AV, Bito H, Bear MF, Jason SD .  Arc restores juvenile plasticity in adult mouse visual cortex .  Proc Natl Acad Sci USA114   9182 - 9187   2017.8Arc restores juvenile plasticity in adult mouse visual cortexReviewed International journal

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  • Kasumi Inokuchi, Fumiaki Imamura, Haruki Takeuchi, Ryang Kim, Hiroyuki Okuno, Hirofumi Nishizumi, Haruhiko Bito, Takefumi Kikusui, Hitoshi Sakano .  Nrp2 is sufficient to instruct circuit formation of mitral-cells to mediate odour-induced attractive social responses .  NATURE COMMUNICATIONS8   15977   2017.7Nrp2 is sufficient to instruct circuit formation of mitral-cells to mediate odour-induced attractive social responses

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    Odour information induces various innate responses that are critical to the survival of the individual and for the species. An axon guidance molecule, Neuropilin 2 (Nrp2), is known to mediate targeting of olfactory sensory neurons (primary neurons), to the posteroventral main olfactory bulb (PV MOB) in mice. Here we report that Nrp2-positive (Nrp2(+)) mitral cells (MCs, second-order neurons) play crucial roles in transmitting attractive social signals from the PV MOB to the anterior part of medial amygdala (MeA). Semaphorin 3F, a repulsive ligand to Nrp2, regulates both migration of Nrp2(+) MCs to the PV MOB and their axonal projection to the anterior MeA. In the MC-specific Nrp2 knockout mice, circuit formation of Nrp2(+) MCs and odour-induced attractive social responses are impaired. In utero, electroporation demonstrates that activation of the Nrp2 gene in MCs is sufficient to instruct their circuit formation from the PV MOB to the anterior MeA.

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  • Hirano Y, Ihara K, Masuda T, Yamamoto T, Iwata I, Takahashi A, Awata H, Nakamura N, Takakura M, Suzuki Y, Horiuchi J, Okuno H, Saitoe M .  Shifting transcriptional machinery is required for long-term memory maintenance and modification in Drosophila mushroom bodies .  NATURE COMMUNICATIONS7   13471   2016.11Shifting transcriptional machinery is required for long-term memory maintenance and modification in Drosophila mushroom bodiesReviewed

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  • Fukuchi M, Nakashima F, Tabuchi A, Shimotori M, Tatsumi S, Okuno H, Bito H, Tsuda M. .  Class I histone deacetylase-mediated repression of the proximal promoter of the activity-regulated cytoskeleton-associated protein gene regulates its response to brain-derived neurotrophic factor .  Journal of Biological Chemistry290   6825 - 6836   2015.5Class I histone deacetylase-mediated repression of the proximal promoter of the activity-regulated cytoskeleton-associated protein gene regulates its response to brain-derived neurotrophic factor

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  • Yamamoto K, Tanei Z, Hashimoto T, Wakabayashi T, Okuno H, Naka Y, Yizhar O, Fenno LE, Fukayama M, Bito H, Cirrito JR, Holtzman DM, Deisseroth K, Iwatsubo T .  Chronic optogenetic activation augments Abeta pathology in a mouse model of Alzheimer disease .  Cell Reports11 ( 6 ) 859 - 865   2015.5Chronic optogenetic activation augments Abeta pathology in a mouse model of Alzheimer diseaseReviewed International journal

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  • Vousden DA, Epp J, Okuno H, Nieman BJ, van Eede M, Dazai J, Ragan T, Bito H, Frankland PW, Lerch JP, Henkelman RM .  Whole-brain mapping of behaviourally induced neural activation in mice .  Brain Struct Funct.220 ( 4 ) 2043 - 2057   2015.4Whole-brain mapping of behaviourally induced neural activation in miceReviewed International journal

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  • Fukuchi M, Tabuchi A, Kuwana Y, Watanabe S, Inoue M, Takasaki I, Izumi H, Tanaka A, Inoue R, Mori H, Komatsu H, Takemori H, Okuno H, Bito H, Tsuda M. .  Neuromodulatory effect of Gαs- or Gαq-coupled G-protein-coupled receptor on NMDA receptor selectively activates the NMDA receptor/Ca2+/calcineurin/cAMP response element-binding protein-regulated transcriptional coactivator 1 pathway to effectively induce brain-derived neurotrophic factor expression in neurons .  Journal of Neuroscience35 ( 14 ) 5606 - 5624   2015.4Neuromodulatory effect of Gαs- or Gαq-coupled G-protein-coupled receptor on NMDA receptor selectively activates the NMDA receptor/Ca2+/calcineurin/cAMP response element-binding protein-regulated transcriptional coactivator 1 pathway to effectively induce brain-derived neurotrophic factor expression in neurons

  • Nonaka M, Kim R, Fukushima H, Sasaki K, Suzuki K, Okamura M, Ishii Y, Kawashima T, Kamijo S, Takemoto-Kimura S, Okuno H, Kida S, Bito H. .  Region-specific activation of CRTC1-CREB signaling mediates long-term fear memory. .  Neuron84 ( 1 ) 92 - 106   2014.10Region-specific activation of CRTC1-CREB signaling mediates long-term fear memory.

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  • Nonaka M, Fujii H, Kim R, Kawashima T, Okuno H, Bito H .  Untangling the two-way signalling route from synapses to the nucleus, and from the nucleus back to the synapses. .  Philos Trans R Soc Lond B Biol Sci.369 ( 1633 ) 20130150   2013.12Untangling the two-way signalling route from synapses to the nucleus, and from the nucleus back to the synapses.Reviewed International journal

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  • Kawashima T, Kitamura K, Suzuki K, Nonaka M, Kamijo S, Takemoto-Kimura S, Kano M, Okuno H, Ohki K, Bito H. .  Functional labeling of neurons and their projections using the synthetic activity-dependent promoter E-SARE. .  Nat Methods10 ( 9 ) 889 - 895   2013.7Functional labeling of neurons and their projections using the synthetic activity-dependent promoter E-SARE.

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  • Mikuni T, Uesaka N, Okuno H, Hirai H, Deisseroth K, Bito H, Kano M. .  Arc/Arg3.1 is a postsynaptic mediator of activity-dependent synapse elimination in the developing cerebellum. .  Neuron78 ( 6 ) 1024 - 1035   2013.6Arc/Arg3.1 is a postsynaptic mediator of activity-dependent synapse elimination in the developing cerebellum.Reviewed International journal

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  • Fujii H, Inoue M, Okuno H, Sano Y, Takemoto-Kimura S, Kitamura K, Kano M, Bito H. .  Nonlinear decoding and asymmetric representation of neuronal input information by CaMKIIα and calcineurin. .  Cell Reports3 ( 4 ) 978 - 987   2013.4Nonlinear decoding and asymmetric representation of neuronal input information by CaMKIIα and calcineurin.

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  • Komatsu T, Johnsson K, Okuno H, Bito H, Inoue T, Nagano T, Urano Y. .  Real-time measurements of protein dynamics using fluorescence activation-coupled protein labeling method. .  J Am Chem Soc133 ( 17 ) 6745 - 6751   2011.5Real-time measurements of protein dynamics using fluorescence activation-coupled protein labeling method.

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  • Sato T, Tokuyama W, Miyashita Y, Okuno H. .  Temporal and spatial dissociation of expression patterns between Zif268 and c-Fos in rat inferior olive during vestibular compensation. .  Neuroreport8 ( 8 ) 1891 - 1895   1997.5Temporal and spatial dissociation of expression patterns between Zif268 and c-Fos in rat inferior olive during vestibular compensation.

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    DOI: 10.1097/00001756-199705260-00020

  • Okuno H, Saffen DW, Miyashita Y. .  Subdivision-specific expression of ZIF268 in the hippocampal formation of the macaque monkey. .  Neuroscience66 ( 4 ) 829 - 845   1995.6Subdivision-specific expression of ZIF268 in the hippocampal formation of the macaque monkey.

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    DOI: 10.1016/0306-4522(94)00619-g

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MISC

  • 【「新しい分子生物学技術を用いた生体機能解析」】化学遺伝学・DREADD法と生体機能解析への応用

    比嘉 なつみ, 奥野 浩行

    LABIO 21   ( 89 )   6 - 8   2023.5

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  • 「新しい分子生物学技術を用いた生体機能解析」1.化学遺伝学・DREADD法と生体機能解析への応用

    比嘉なつみ, 奥野浩行

    Labio 21   ( 89 )   2023

  • Changes in calcium responses overtime in single neurons in the primary somatosensory cortex in response to pain stimuli revealed by in vivo calcium imaging using a head-mounted fluorescence microscope

    大徳晋久, 比嘉なつみ, 城山優治, 奥野浩行, 松永明

    日本麻酔科学会学術集会(Web)   70th   2023

  • Changes in calcium responses during isoflurane inhalation anesthesia in the mouse hippocampus using a miniaturized fluorescence microscope

    大江将軍, 大徳晋久, 城山優治, 上村裕一, 奥野浩行, 松永明

    日本麻酔科学会学術集会(Web)   70th   2023

  • Gastrin-releasing peptideは急性ストレス下で,amygdalostriatal transition areaを通じて恐怖学習を調節する

    城山優治, 城山優治, 後藤史子, 小川糸音, 吉田進昭, 尾藤晴彦, 真鍋俊也, 奥野浩行, 奥野浩行

    日本生化学会大会(Web)   95th   2022

  • Gastrin-releasing peptideはamygdalostriatal transition areaを活性化させ,ストレス負荷時の恐怖学習を調節する

    後藤史子, 城山優治, 城山優治, 小川糸音, 奥野浩行, 吉田進昭, 尾藤晴彦, 真鍋俊也

    日本神経化学会大会抄録集(Web)   65th   2022

  • 2光子生体脳スパインイメージングで明らかになった恐怖条件付け記憶回路におけるスパイン新生

    湊原圭一郎, 湊原圭一郎, 湊原圭一郎, 大波壮一郎, 大波壮一郎, 岡部繁男, 尾藤晴彦, 奥野浩行, 奥野浩行

    日本神経化学会大会抄録集(Web)   65th   2022

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Presentations

  • 湊原 圭一郎, 鈴木 穣, 菅野 純夫, 尾藤 晴彦, 奥野 浩行   認知活動により活性化される前初期遺伝子Arc発現海馬神経細胞におけるスパイン形態解析およびトランスクリプトーム解析  

    日本生化学会大会プログラム・講演要旨集  2018.9  (公社)日本生化学会

  • 中原 真由美, 向原 桂香, 大江 将軍, 城山 優治, 奥野 浩行, 上村 裕一   術後認知機能障害に関与する大脳神経回路同定の試み  

    日本集中治療医学会雑誌  2020.9  (一社)日本集中治療医学会

  • 奥野 浩行, 荒木 杏菜, 湊原 圭一郎   若手研究者が取り組む脳科学研究の最新技術と創薬応用 シナプス活動依存的遺伝子発現に基づく記憶関連細胞標識法の開発と応用  

    日本薬学会年会要旨集  2017.3  (公社)日本薬学会

  • 田渕 明子, 福地 守, 菊池 啓悦, 久保 友喜美, 庄司 しずく, 袴田 知之, 田中 拓郎, 佐藤 夏美, 石橋 悠太, 金田 真理彩, 阪上 洋行, 尾藤 晴彦, 奥野 浩行, 大塚 稔久, 津田 正明   細胞骨格により制御される転写と細胞機能:細胞形質転換と疾患 神経細胞におけるSRFコファクターMKL(MRTF)による遺伝子発現制御と形態制御  

    生命科学系学会合同年次大会  2017.12  生命科学系学会合同年次大会運営事務局

  • 金子 雅規, 伊藤 政之, 山下 真梨子, 奥野 浩行, 阿部 学, 山崎 真弥, 夏目 里恵, 貝塚 利恵, 崎村 建司, 星野 幹雄, 三品 昌美, 林 崇   大脳異常興奮の抑制に関わるAMPA受容体パルミトイル化依存的なシナプス機能制御  

    日本生物学的精神医学会・日本神経精神薬理学会合同年会プログラム・抄録集  2017.9  日本生物学的精神医学会・日本神経精神薬理学会

  • 城山 優治, 中原 真由美, 大江 将軍, 坂口 茜, 黒江 那彩, 上村 裕一, 奥野 浩行   マウスにおける全身麻酔下手術による扁桃体の神経活動亢進と記憶障害  

    日本生化学会大会プログラム・講演要旨集  2020.9  (公社)日本生化学会

  • Itoh Masayuki, Yamashita Mariko, Okuno Hiroyuki, Abe Manabu, Yamazaki Maya, Natsume Rie, Kaizuka Toshie, Sakimura Kenji, Hoshino Mikio, Mishina Masayoshi, Hayashi Takashi   パルミトイル化によるAMPA型グルタミン受容体の制御は発作感受性を調節する(Palmitoylation-dependent regulation of the AMPA receptor modulates seizure susceptibility)  

    Journal of Pharmacological Sciences  2017.3  (公社)日本薬理学会

  • Okuno Hiroyuki, Ishii Yuichiro, Endo Toshihiro, Minatohara Keiichiro, Bito Haruhiko   シナプス生理学の解明 シナプス機能の最先端研究 ArcによるAMPA受容体の表面動力学および認知処理の調節(Illumination of Synapse Physiology: Frontier researches of synaptic functions Regulation of surface dynamics of AMPA receptors and cognitive processes by Arc)  

    The Journal of Physiological Sciences  2018.3  (一社)日本生理学会

  • Ishii Yuichiro, Okuno Hiroyuki, Fujii Hajime, Bito Haruhiko   グルタミン酸シナプスの分子・細胞神経生物学 表面グルタミン酸受容体発現の長期増強の単一シナプスイメージング(Molecular and cellular neurobiology of glutamatergic synapse Imaging long-term potentiation of surface glutamate receptors at single synapses)  

    The Journal of Physiological Sciences  2017.3  (一社)日本生理学会

  • 城山 優治, 奥野 浩行, 内藤 眞, 尾藤 晴彦, 児玉 龍彦, 真鍋 俊也, 穴井 元暢   てんかん発症に至るまでの海馬歯状回 APE欠損をモデルとして  

    日本生化学会大会プログラム・講演要旨集  2019.9  (公社)日本生化学会

  • Bito Haruhiko, Ishii Yuichiro, Okuno Hiroyuki   Synaptic tagging and capture hypothesisに対する新しい分子的洞察 逆synaptic tagging 活動誘発性Arcを捕捉するために不活性シナプスを標的とするメカニズム(New molecular insights into the synaptic tagging and capture hypothesis Inverse synaptic tagging: an inactive synapse-targeted mechanism to capture activity-induced Arc)  

    The Journal of Physiological Sciences  2019.6  (一社)日本生理学会

  • Okuno Hiroyuki, Araki Anna, Minatohara Keiichiro, Bito Haruhiko, Imayoshi Itaru   Arc-based functional labelingにより明らかとなったマウス海馬の遠隔記憶痕跡(Remote memory traces in the mouse hippocampus revealed by Arc-based functional labeling)  

    The Journal of Physiological Sciences  2019.6  (一社)日本生理学会

  • 城山 優治, 大西 哲生, 有馬 史子, 門田 満隆, 尾藤 晴彦, 吉川 武男, 真鍋 俊也, 奥野 浩行   Ldb2 modulates synaptic function and fear learning by regulating Arc expression in the lateral nucleus of the amygdala(和訳中)  

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • 城山 優治, 後藤 史子, 小川 糸音, 吉田 進昭, 尾藤 晴彦, 真鍋 俊也, 奥野 浩行   Gastrin-releasing peptideは急性ストレス下で、amygdalostriatal transition areaを通じて恐怖学習を調節する  

    日本生化学会大会プログラム・講演要旨集  2022.11  (公社)日本生化学会

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  • 城山 優治, 大西 哲生, 有馬 史子, 門田 満隆, 尾藤 晴彦, 吉川 武男, 真鍋 俊也, 奥野 浩行   Ldb2は扁桃体側核におけるArcの発現を制御することでシナプス機能と恐怖条件づけ学習を制御する(Ldb2 modulates synaptic function and fear learning by regulating Arc expression in the lateral nucleus of the amygdala)  

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • 城山 優治, 大西 哲生, 有馬 史子, 門田 満隆, 尾藤 晴彦, 吉川 武男, 真鍋 俊也, 奥野 浩行   Ldb2は扁桃体側核におけるArcの発現を制御することでシナプス機能と恐怖条件づけ学習を制御する(Ldb2 modulates synaptic function and fear learning by regulating Arc expression in the lateral nucleus of the amygdala)  

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • 城山 優治, 大西 哲生, 有馬 史子, 門田 満隆, 尾藤 晴彦, 吉川 武男, 真鍋 俊也, 奥野 浩行   Ldb2は扁桃体側核におけるArcの発現を制御することでシナプス機能と恐怖条件づけ学習を制御する(Ldb2 modulates synaptic function and fear learning by regulating Arc expression in the lateral nucleus of the amygdala)  

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • 城山 優治, 大西 哲生, 有馬 史子, 門田 満隆, 尾藤 晴彦, 吉川 武男, 真鍋 俊也, 奥野 浩行   Ldb2は扁桃体側核におけるArcの発現を制御することでシナプス機能と恐怖条件づけ学習を制御する(Ldb2 modulates synaptic function and fear learning by regulating Arc expression in the lateral nucleus of the amygdala)  

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • 城山 優治, 大西 哲生, 有馬 史子, 門田 満隆, 尾藤 晴彦, 吉川 武男, 真鍋 俊也, 奥野 浩行   Ldb2は扁桃体側核におけるArcの発現を制御することでシナプス機能と恐怖条件づけ学習を制御する(Ldb2 modulates synaptic function and fear learning by regulating Arc expression in the lateral nucleus of the amygdala)  

    日本生化学会大会プログラム・講演要旨集  2021.11  (公社)日本生化学会

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  • Kaneko Tatsuroh, Oura Asuka, Kusumoto-Yoshida Ikue, Kanekura Takuro, Okuno Hiroyuki, Kuwaki Tomoyuki, Kashiwadani Hideki   痒み研究の新展開 痒みと痛みのneural processingにおけるオレキシン神経の役割(New insights into itch research The contrasting roles of orexin neurons in itch and pain neural processing)  

    The Journal of Physiological Sciences  2024.5  (一社)日本生理学会

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Research Projects

  • 周術期免疫抑制が神経活動へ及ぼす影響と術後認知障害の関連の解明

    Grant number:22K09088  2022.4 - 2025.3

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    長谷川 麻衣子, 内田 寛治, 奥野 浩行, 住谷 昌彦

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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  • 雄マウス求愛発声と性的動機づけを担うドーパミン神経投射経路の同定

    Grant number:21K03144  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    菅野 康太, 奥野 浩行

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    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

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  • In vitro, in vivo functions and involvement in schizophrenia of lipid modifying enzyme that shows activity-dependent expression

    Grant number:21K06375  2021.4 - 2024.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (C)  Grant-in-Aid for Scientific Research (C)

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    Grant amount:\4290000 ( Direct Cost: \3300000 、 Indirect Cost:\990000 )

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  • Verification study and therapeutical utilization for implementation of real time monitoring in oral cancer

    Grant number:21H03143  2021.4 - 2024.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)  Grant-in-Aid for Scientific Research (B)

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    Grant amount:\17290000 ( Direct Cost: \13300000 、 Indirect Cost:\3990000 )

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  • インビボ神経イメージングによる敗血症関連脳症に関与する大脳神経回路同定と治療戦略

    Grant number:21K09077  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    中原 真由美, 新山 修平, 奥野 浩行, 城山 優治

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    Grant amount:\4160000 ( Direct Cost: \3200000 、 Indirect Cost:\960000 )

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  • 神経応答-遺伝子発現変換を起点とした記憶痕跡形成過程の情報動態解析

    Grant number:20H05068  2020.4 - 2022.3

    日本学術振興会  科学研究費助成事業 新学術領域研究(研究領域提案型)  新学術領域研究(研究領域提案型)

    奥野 浩行

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    Grant amount:\5720000 ( Direct Cost: \4400000 、 Indirect Cost:\1320000 )

    学習時に前初期遺伝子発現を伴う活性化神経細胞は記憶痕跡(エングラム)として機能する。しかしながら、記憶獲得時の神経活動が前初期遺伝子現を誘起する機構は不明であり、さらには、前初期遺伝子発現が記憶痕跡の性質獲得に寄与する分子機構についてもほとんど理解が進んでいない。これらの知見を得るため、本研究では学習時や記憶想起時の神経応答パターンとそれに続く前初期遺伝子発現をマウスを用いて同一個体で連続的に解析する。この目的のため、本年度はまず緑色光励起および赤色蛍光の画像取得を可能とする頭部搭載型顕微鏡の作成を行った。作成された顕微鏡は感度面では緑色蛍光取得システムには及ばないものの前初期遺伝子発現レポーターを観察するためには十分な性能を有した。また、緑色蛍光の遺伝子発現レポーターと組み合わせて使用できるように赤色蛍光のカルシウムセンサー(XCaMP-R)を研究室に導入し、これを神経細胞特異的プロモーター(CaMK2プロモーターやSynapsinIプロモーター)下で発現するコンストラクトを作成し、アデノ随伴ウイルス(AAV)ベクターにパッケージングした。同様に新たな前初期遺伝子発現レポーターを構築し、AAVベクターにパッケージング、精製を行った。さらに、前初期遺伝子発現に続く後期遺伝子発現の網羅的転写産物解析を行うにあたり、これまで未解析の時点データを得るため新たに可塑性誘発刺激後の様々なタイミングで海馬歯状回顆粒細胞を採取し、そこからRNA-Seqライブラリーを作成した。

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  • オプトジェネティクスをもちいた麻酔薬による術後認知障害の機序の解明

    Grant number:19K09355  2019.4 - 2022.3

    日本学術振興会  科学研究費助成事業 基盤研究(C)  基盤研究(C)

    向原 桂香, 長谷川 麻衣子, 奥野 浩行

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    Grant amount:\4420000 ( Direct Cost: \3400000 、 Indirect Cost:\1020000 )

    神経活動レポーター遺伝子の脳内注入:定位脳手術により、前頭前野、海馬、扁桃体に注入することや、レンズプローブ植込み神経活動レポーターの発現評価:前頭前野および海馬、または偏桃体上にグリンレンズプローブを植込み、神経活動レポーターの局在と発現量を小型統合蛍光顕微鏡をもちいて評価することの実験系作成をおこなった。全身麻酔下手術侵襲中の神経活動評価:2%イソフルラン± NMDA antagonist (MK801 0.05 mg/kg i.p.)投与下に術後痛モデルを作成した。切開前後の前頭前野、海馬における神経活動による細胞内カルシウム応答GCaMP6f、また、活動依存的な遺伝子発現をArc-GFPレポーターによってリアルタイムに測定する。4術後記憶・認知行動評価と神経活動の相関:術前に学習をおこない、術後にマウスに音を聴かせ条件刺激 (音)のみですくみ反応が出現するか評価し、恐怖記憶への麻酔下手術侵襲の影響を明らかにする。
    5脳マトリックス間の相互作用:前頭前野―海馬―扁桃体の連関はArc-Pro-Venus-Pestトランスジェニックマウスをもちいて検証した。手術前後のArc依存性Venus発現神経細胞を免疫組織染色によって評価した。オプトジェネティクスをもちいた術中Arc+neuron操作と術後恐怖関連痛み行動の検討:全身麻酔中にArc-ChR2あるいはArch-T-Arc neuronをそれぞれ青色光、緑色光を当てて活性化もしくは抑制し、この影響を条件付け恐怖行動実験によって評価する過程にある。

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  • 機能的回路多重標識と転写産物プロファイリングによる脳多領野記憶痕跡モデルの検証

    2018 - 2019

    文部科学省  新学術領域研究(研究領域提案型) 

    奥野 浩行

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  • シナプス可塑性関連因子Arcによる認知機能調節機構の解明

    2015 - 2018

    文部科学省  科学研究費補助金(基盤研究(B)) 

    奥野 浩行

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