Updated on 2024/04/02

写真a

 
WAKE Hiroaki
 
Organization
Institutes of Innovation for Future Society Professor
Graduate School of Medicine Professor
Graduate School
Graduate School of Medicine
Title
Professor

Degree 1

  1. 医学博士 ( 2007.3   名古屋市立大学 ) 

Research Interests 6

  1. 脳梗塞

  2. 神経-グリア相関

  3. 生体

  4. ミクログリア

  5. シナプス

  6. 2光子励起顕微鏡

Research Areas 5

  1. Life Science / Clinical pharmacy

  2. Life Science / Neuroscience-general

  3. Life Science / Neuroscience-general

  4. Life Science / Neuroscience-general

  5. Life Science / Physiology

Research History 2

  1. Nagoya University   Professor

    2019.11

  2. Kobe University   Graduate School of Medicine   Professor

    2016.4 - 2019.10

Education 1

  1. 名古屋市立大学 大学院医学研究科博士課程生体情報機能制御医学専攻修了

    - 2007.3

Awards 1

  1. 神戸大学優秀若手研究賞・学長賞

    2017   神戸大学   高次脳機能の解明と操作に光を当てる

    WAKE HIROAKI

 

Papers 59

  1. Astrocytic NKCC1 inhibits seizures by buffering Cl(-) and antagonizing neuronal NKCC1 at GABAergic synapses.

    Nguyen TD, Ishibashi M, Sinha AS, Watanabe M, Kato D, Horiuchi H, Wake H, Fukuda A

    Epilepsia   Vol. 64 ( 12 ) page: 3389 - 3403   2023.12

     More details

    Language:English   Publisher:Epilepsia  

    Objective: A pathological excitatory action of the major inhibitory neurotransmitter γ-aminobutyric acid (GABA) has been observed in epilepsy. Blocking the Cl− importer NKCC1 with bumetanide is expected to reduce the neuronal intracellular Cl− concentration ([Cl−]i) and thereby attenuate the excitatory GABA response. Accordingly, several clinical trials of bumetanide for epilepsy were conducted. Although NKCC1 is expressed in both neurons and glial cells, an involvement of glial NKCC1 in seizures has not yet been reported. Astrocytes maintain high [Cl−]i with NKCC1, and this gradient promotes Cl− efflux via the astrocytic GABAA receptor (GABAAR). This Cl− efflux buffers the synaptic cleft Cl− concentration to maintain the postsynaptic Cl− gradient during intense firing of GABAergic neurons, thereby sustaining its inhibitory action during seizure. In this study, we investigated the function of astrocytic NKCC1 in modulating the postsynaptic action of GABA in acute seizure models. Methods: We used the astrocyte-specific conditional NKCC1 knockout (AstroNKCC1KO) mice. The seizurelike events (SLEs) in CA1 pyramidal neurons were triggered by tetanic stimulation of stratum radiatum in acute hippocampus slices. The SLE underlying GABAAR-mediated depolarization was evaluated by applying the GABAAR antagonist bicuculline. The pilocarpine-induced seizure in vivo was monitored in adult mice by the Racine scale. The SLE duration and tetanus stimulation intensity threshold and seizure behavior in AstroNKCC1KO mice and wild-type (WT) mice were compared. Results: The AstroNKCC1KO mice were prone to seizures with lower threshold and longer duration of SLEs and larger GABAAR-mediated depolarization underlying the SLEs, accompanied by higher Racine-scored seizures. Bumetanide reduced these indicators of seizure in AstroNKCC1KO mice (which still express neuronal NKCC1), but not in the WT, both in vitro and in vivo. Significance: Astrocytic NKCC1 inhibits GABA-mediated excitatory action during seizures, whereas neuronal NKCC1 has the converse effect, suggesting opposing actions of bumetanide on these cells.

    DOI: 10.1111/epi.17784

    Scopus

    PubMed

  2. Regulation of lipid synthesis in myelin modulates neural activity and is required for motor learning

    Kato, D; Aoyama, Y; Nishida, K; Takahashi, Y; Sakamoto, T; Takeda, I; Tatematsu, T; Go, S; Saito, Y; Kunishima, S; Cheng, JL; Hou, LN; Tachibana, Y; Sugio, S; Kondo, R; Eto, F; Sato, S; Moorhouse, AJ; Yao, IKK; Kadomatsu, K; Setou, M; Wake, H

    GLIA   Vol. 71 ( 11 ) page: 2591 - 2608   2023.11

     More details

    Language:English   Publisher:GLIA  

    Brain function relies on both rapid electrical communication in neural circuitry and appropriate patterns or synchrony of neural activity. Rapid communication between neurons is facilitated by wrapping nerve axons with insulation by a myelin sheath composed largely of different lipids. Recent evidence has indicated that the extent of myelination of nerve axons can adapt based on neural activity levels and this adaptive myelination is associated with improved learning of motor tasks, suggesting such plasticity may enhance effective learning. In this study, we examined whether another aspect of myelin plasticity—changes in myelin lipid synthesis and composition—may also be associated with motor learning. We combined a motor learning task in mice with in vivo two-photon imaging of neural activity in the primary motor cortex (M1) to distinguish early and late stages of learning and then probed levels of some key myelin lipids using mass spectrometry analysis. Sphingomyelin levels were elevated in the early stage of motor learning while galactosylceramide levels were elevated in the middle and late stages of motor learning, and these changes were correlated across individual mice with both learning performance and neural activity changes. Targeted inhibition of oligodendrocyte-specific galactosyltransferase expression, the enzyme that synthesizes myelin galactosylceramide, impaired motor learning. Our results suggest regulation of myelin lipid composition could be a novel facet of myelin adaptations associated with learning.

    DOI: 10.1002/glia.24441

    Web of Science

    Scopus

    PubMed

  3. Microglial process dynamics depend on astrocyte and synaptic activity.

    Ikegami A, Kato D, Wake H

    Nagoya journal of medical science   Vol. 85 ( 4 ) page: 772 - 778   2023.11

     More details

    Language:English   Publisher:Nagoya Journal of Medical Science  

    Microglial processes survey the brain parenchyma, but it is unknown whether this process is influenced by the cell activity of nearby microglia under physiological conditions. Herein, we showed that microglial process dynamics differ when facilitated by astrocytic activity and pre-synaptic activity. The results revealed distinct microglial process dynamics associated with the activity of other brain cells.

    DOI: 10.18999/nagjms.85.4.772

    Scopus

    PubMed

  4. Regulation of lipid synthesis in myelin modulates neural activity and is required for motor learning Reviewed

    Daisuke Kato, Yuki Aoyama, Kazuki Nishida, Yutaka Takahashi, Takumi Sakamoto, Ikuko Takeda, Tsuyako Tatematsu, Shiori Go, Yutaro Saito, Shiho Kunishima, Jinlei Cheng, Lingnan Hou, Yoshihisa Tachibana, Shouta Sugio, Reon Kondo, Fumihiro Eto, Shumpei Sato, Andrew J Moorhouse, Ikuko Yao, Kenji Kadomatsu, Mitsutoshi Setou, Hiroaki Wake

    Glia     2023.7

     More details

    Publishing type:Research paper (scientific journal)   Publisher:Wiley  

    Abstract

    Brain function relies on both rapid electrical communication in neural circuitry and appropriate patterns or synchrony of neural activity. Rapid communication between neurons is facilitated by wrapping nerve axons with insulation by a myelin sheath composed largely of different lipids. Recent evidence has indicated that the extent of myelination of nerve axons can adapt based on neural activity levels and this adaptive myelination is associated with improved learning of motor tasks, suggesting such plasticity may enhance effective learning. In this study, we examined whether another aspect of myelin plasticity—changes in myelin lipid synthesis and composition—may also be associated with motor learning. We combined a motor learning task in mice with in vivo two‐photon imaging of neural activity in the primary motor cortex (M1) to distinguish early and late stages of learning and then probed levels of some key myelin lipids using mass spectrometry analysis. Sphingomyelin levels were elevated in the early stage of motor learning while galactosylceramide levels were elevated in the middle and late stages of motor learning, and these changes were correlated across individual mice with both learning performance and neural activity changes. Targeted inhibition of oligodendrocyte‐specific galactosyltransferase expression, the enzyme that synthesizes myelin galactosylceramide, impaired motor learning. Our results suggest regulation of myelin lipid composition could be a novel facet of myelin adaptations associated with learning.

    DOI: 10.1002/glia.24441.

  5. Microglia enable cross-modal plasticity by removing inhibitory synapses. International journal

    Akari Hashimoto, Nanami Kawamura, Etsuko Tarusawa, Ikuko Takeda, Yuki Aoyama, Nobuhiko Ohno, Mio Inoue, Mai Kagamiuchi, Daisuke Kato, Mami Matsumoto, Yoshihiro Hasegawa, Junichi Nabekura, Anne Schaefer, Andrew J Moorhouse, Takeshi Yagi, Hiroaki Wake

    Cell reports   Vol. 42 ( 5 ) page: 112383 - 112383   2023.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Cross-modal plasticity is the repurposing of brain regions associated with deprived sensory inputs to improve the capacity of other sensory modalities. The functional mechanisms of cross-modal plasticity can indicate how the brain recovers from various forms of injury and how different sensory modalities are integrated. Here, we demonstrate that rewiring of the microglia-mediated local circuit synapse is crucial for cross-modal plasticity induced by visual deprivation (monocular deprivation [MD]). MD relieves the usual inhibition of functional connectivity between the somatosensory cortex and secondary lateral visual cortex (V2L). This results in enhanced excitatory responses in V2L neurons during whisker stimulation and a greater capacity for vibrissae sensory discrimination. The enhanced cross-modal response is mediated by selective removal of inhibitory synapse terminals on pyramidal neurons by the microglia in the V2L via matrix metalloproteinase 9 signaling. Our results provide insights into how cortical circuits integrate different inputs to functionally compensate for neuronal damage.

    DOI: 10.1016/j.celrep.2023.112383

    Web of Science

    Scopus

    PubMed

  6. KCC2 downregulation after sciatic nerve injury enhances motor function recovery.

    Cheung DL, Toda T, Narushima M, Eto K, Takayama C, Ooba T, Wake H, Moorhouse AJ, Nabekura J

    Scientific reports   Vol. 13 ( 1 ) page: 7871   2023.5

     More details

    Language:English   Publisher:Scientific Reports  

    Injury to mature neurons induces downregulated KCC2 expression and activity, resulting in elevated intracellular [Cl−] and depolarized GABAergic signaling. This phenotype mirrors immature neurons wherein GABA-evoked depolarizations facilitate neuronal circuit maturation. Thus, injury-induced KCC2 downregulation is broadly speculated to similarly facilitate neuronal circuit repair. We test this hypothesis in spinal cord motoneurons injured by sciatic nerve crush, using transgenic (CaMKII-KCC2) mice wherein conditional CaMKIIα promoter-KCC2 expression coupling selectively prevents injury-induced KCC2 downregulation. We demonstrate, via an accelerating rotarod assay, impaired motor function recovery in CaMKII-KCC2 mice relative to wild-type mice. Across both cohorts, we observe similar motoneuron survival and re-innervation rates, but differing post-injury reorganization patterns of synaptic input to motoneuron somas—for wild-type, both VGLUT1-positive (excitatory) and GAD67-positive (inhibitory) terminal counts decrease; for CaMKII-KCC2, only VGLUT1-positive terminal counts decrease. Finally, we recapitulate the impaired motor function recovery of CaMKII-KCC2 mice in wild-type mice by administering local spinal cord injections of bicuculline (GABAA receptor blockade) or bumetanide (lowers intracellular [Cl−] by NKCC1 blockade) during the early post-injury period. Thus, our results provide direct evidence that injury-induced KCC2 downregulation enhances motor function recovery and suggest an underlying mechanism of depolarizing GABAergic signaling driving adaptive reconfiguration of presynaptic GABAergic input.

    DOI: 10.1038/s41598-023-34701-y

    Scopus

    PubMed

  7. Distinct features of two lipid droplets types in cell nuclei from patients with liver diseases

    Norihiro Imai, Yuki Ohsaki, Jinglei Cheng, Jingjing Zhang, Fumitaka Mizuno, Taku Tanaka, Shinya Yokoyama, Kenta Yamamoto, Takanori Ito, Yoji Ishizu, Takashi Honda, Masatoshi Ishigami, Hiroaki Wake, Hiroki Kawashima

    Scientific Reports   Vol. 13 ( 1 ) page: 6851   2023.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media {LLC}  

    <jats:title>Abstract</jats:title><jats:p>Lipid droplets (LDs) have been observed in the nuclei of hepatocytes; however, their significance in liver disease remains unresolved. Our purpose was to explore the pathophysiological features of intranuclear LDs in liver diseases. We included 80 patients who underwent liver biopsies; the specimens were dissected and fixed for electron microscopy analysis. Depending on the presence of adjacent cytoplasmic invagination of the nuclear membrane, LDs in the nuclei were classified into two types: nucleoplasmic LDs (nLDs) and cytoplasmic LD invagination with nucleoplasmic reticulum (cLDs in NR). nLDs were found in 69% liver samples and cLDs in NR were found in 32%; no correlation was observed between the frequencies of the two LD types. nLDs were frequently found in hepatocytes of patients with nonalcoholic steatohepatitis, whereas cLDs in NR were absent from the livers of such patients. Further, cLDs in NR were often found in hepatocytes of patients with lower plasma cholesterol level. This indicates that nLDs do not directly reflect cytoplasmic lipid accumulation and that formation of cLDs in NR is inversely correlated to the secretion of very low-density lipoproteins. Positive correlations were found between the frequencies of nLDs and endoplasmic reticulum (ER) luminal expansion, suggesting that nLDs are formed in the nucleus upon ER stress. This study unveiled the presence of two distinct nuclear LDs in various liver diseases. </jats:p>

    DOI: 10.1038/s41598-023-33977-4

    Web of Science

    Scopus

    PubMed

  8. CD206+ macrophages transventricularly infiltrate the early embryonic cerebral wall to differentiate into microglia

    Yuki Hattori, Daisuke Kato, Futoshi Murayama, Sota Koike, Hisa Asai, Ayato Yamasaki, Yu Naito, Ayano Kawaguchi, Hiroyuki Konishi, Marco Prinz, Takahiro Masuda, Hiroaki Wake, Takaki Miyata

    Cell Reports   Vol. 42 ( 2 ) page: 112092 - 112092   2023.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.celrep.2023.112092

    Web of Science

    Scopus

    PubMed

  9. Myelinated axon as a plastic cable regulating brain functions. International journal

    Shouta Sugio, Daisuke Kato, Hiroaki Wake

    Neuroscience research   Vol. 187   page: 45 - 51   2023.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Each oligodendrocyte (OC) forms myelin approximately in around 10 different axons to coordinate information transfer by regulating conduction velocity in the central nervous system (CNS). In the classical view, myelin has been considered a static structure that rarely turns over under healthy conditions because myelin tightly holds axons by their laminar complex structure. However, in recent decades, the classical views of static myelin have been renewed with pioneering studies that showed plastic changes in myelin throughout life with new experiences, such as the acquisition of new motor skills and the formation of memory. These changes in myelin regulate conduction velocity to optimize the temporal pattern of neuronal circuit activity among distinct brain regions associated with skill learning and memory. Here, we introduce pioneering studies and discuss the implications of plastic myelin on neural circuits and brain function.

    DOI: 10.1016/j.neures.2022.11.002

    Web of Science

    Scopus

    PubMed

  10. Holographic microscope illuminates brain activity

    Kato D., Quan X., Matoba O., Wake H.

    Proceedings of SPIE - The International Society for Optical Engineering   Vol. 12608   2023

     More details

    Publisher:Proceedings of SPIE - The International Society for Optical Engineering  

    Optical bioimaging and optogenetics have recently made it possible to control brain functions. However, these methods have limitations, such as the inability to regulate neural activity with a high spatiotemporal resolution. To address this limitation, we have developed microscope for biological applications by integrating optogenetics and digital holographic technology. This provides precise spatiotemporal information about neural activity.

    DOI: 10.1117/12.3008169

    Scopus

  11. Activity-dependent oligodendrocyte calcium dynamics and their changes in Alzheimer's disease. International journal

    Kenji Yoshida, Daisuke Kato, Shouta Sugio, Ikuko Takeda, Hiroaki Wake

    Frontiers in cellular neuroscience   Vol. 17   page: 1154196 - 1154196   2023

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Oligodendrocytes (OCs) form myelin around axons, which is dependent on neuronal activity. This activity-dependent myelination plays a crucial role in training and learning. Previous studies have suggested that neuronal activity regulates proliferation and differentiation of oligodendrocyte precursor cells (OPCs) and myelination. In addition, deficient activity-dependent myelination results in impaired motor learning. However, the functional response of OC responsible for neuronal activity and their pathological changes is not fully elucidated. In this research, we aimed to understand the activity-dependent OC responses and their different properties by observing OCs using in vivo two-photon microscopy. We clarified that the Ca2+ activity in OCs is neuronal activity dependent and differentially regulated by neurotransmitters such as glutamate or adenosine triphosphate (ATP). Furthermore, in 5-month-old mice models of Alzheimer's disease, a period before the appearance of behavioral abnormalities, the elevated Ca2+ responses in OCs are ATP dependent, suggesting that OCs receive ATP from damaged tissue. We anticipate that our research will help in determining the correct therapeutic strategy for neurodegenerative diseases beyond the synapse.

    DOI: 10.3389/fncel.2023.1154196

    Scopus

    PubMed

  12. [Microglial regulation of brain function and pathological changes].

    Hiroaki Wake, Akari Hashimoto, Daisuke Kato, Ikuko Takeda

    Nihon yakurigaku zasshi. Folia pharmacologica Japonica   Vol. 158 ( 5 ) page: 359 - 361   2023

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)   Publisher:The Japanese Pharmacological Society  

    Microglia are the only immune cells in the central nervous system. It has been shown that microglia actively regulate the number of neurons by participating in the cell death of neural stem cells during development and maturation. In addition, recent optical techniques have enabled in vivo imaging, which has revealed the function of microglia on synapses. Microglia regularly monitor synaptic activity and remove synapses that show abnormal activity in the event of brain infarction or other disorders. During development, microglia contribute to the formation of immature synapses by contacting dendrites during early synapse formation, and they are also involved in the de-synaptic process by selectively removing weakly active synapses through the use of classical complement cascade signaling. Furthermore, these abnormalities are known to contribute to the development of autism during development and to the development of Alzheimer's disease during maturation. In addition to this, microglia also contribute to plastic changes in synapses during the learning process in maturation. Furthermore, by modifying synaptic activity, microglia are known to be involved in changes in the activity of neuronal circuits. In addition to these synaptic functions, microglia are also known to be involved in the permeability of the blood-brain barrier. In this chapter, these functions will be summarized and discussed.

    DOI: 10.1254/fpj.23010

    Scopus

    PubMed

    CiNii Research

  13. Microglia states and nomenclature: A field at its crossroads. International journal

    Rosa C Paolicelli, Amanda Sierra, Beth Stevens, Marie-Eve Tremblay, Adriano Aguzzi, Bahareh Ajami, Ido Amit, Etienne Audinat, Ingo Bechmann, Mariko Bennett, Frederick Bennett, Alain Bessis, Knut Biber, Staci Bilbo, Mathew Blurton-Jones, Erik Boddeke, Dora Brites, Bert Brône, Guy C Brown, Oleg Butovsky, Monica J Carson, Bernardo Castellano, Marco Colonna, Sally A Cowley, Colm Cunningham, Dimitrios Davalos, Philip L De Jager, Bart de Strooper, Adam Denes, Bart J L Eggen, Ukpong Eyo, Elena Galea, Sonia Garel, Florent Ginhoux, Christopher K Glass, Ozgun Gokce, Diego Gomez-Nicola, Berta González, Siamon Gordon, Manuel B Graeber, Andrew D Greenhalgh, Pierre Gressens, Melanie Greter, David H Gutmann, Christian Haass, Michael T Heneka, Frank L Heppner, Soyon Hong, David A Hume, Steffen Jung, Helmut Kettenmann, Jonathan Kipnis, Ryuta Koyama, Greg Lemke, Marina Lynch, Ania Majewska, Marzia Malcangio, Tarja Malm, Renzo Mancuso, Takahiro Masuda, Michela Matteoli, Barry W McColl, Veronique E Miron, Anna Victoria Molofsky, Michelle Monje, Eva Mracsko, Agnes Nadjar, Jonas J Neher, Urte Neniskyte, Harald Neumann, Mami Noda, Bo Peng, Francesca Peri, V Hugh Perry, Phillip G Popovich, Clare Pridans, Josef Priller, Marco Prinz, Davide Ragozzino, Richard M Ransohoff, Michael W Salter, Anne Schaefer, Dorothy P Schafer, Michal Schwartz, Mikael Simons, Cody J Smith, Wolfgang J Streit, Tuan Leng Tay, Li-Huei Tsai, Alexei Verkhratsky, Rommy von Bernhardi, Hiroaki Wake, Valérie Wittamer, Susanne A Wolf, Long-Jun Wu, Tony Wyss-Coray

    Neuron   Vol. 110 ( 21 ) page: 3458 - 3483   2022.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Microglial research has advanced considerably in recent decades yet has been constrained by a rolling series of dichotomies such as "resting versus activated" and "M1 versus M2." This dualistic classification of good or bad microglia is inconsistent with the wide repertoire of microglial states and functions in development, plasticity, aging, and diseases that were elucidated in recent years. New designations continuously arising in an attempt to describe the different microglial states, notably defined using transcriptomics and proteomics, may easily lead to a misleading, although unintentional, coupling of categories and functions. To address these issues, we assembled a group of multidisciplinary experts to discuss our current understanding of microglial states as a dynamic concept and the importance of addressing microglial function. Here, we provide a conceptual framework and recommendations on the use of microglial nomenclature for researchers, reviewers, and editors, which will serve as the foundations for a future white paper.

    DOI: 10.1016/j.neuron.2022.10.020

    Web of Science

    Scopus

    PubMed

  14. 大脳皮質体性感覚野における疼痛で誘発される局所神経回路の活動制御は急性疼痛に対する新たな治療標的になりうる

    岡田 卓也, 野村 有紀, 小幡 典彦, 加藤 大輔, 和氣 弘明, 溝渕 知司

    麻酔   Vol. 71 ( 増刊 ) page: S163 - S170   2022.11

     More details

    Language:Japanese   Publisher:克誠堂出版(株)  

    急性疼痛時や疼痛モデルマウス作製前後における大脳皮質第一次体性感覚野(S1)神経細胞の自発活動や各細胞間の活動相関性および機能的結合の経時変化を検証した。6~8週齢の雄性C57BL/6マウスおよび雄性parvalbumin-Creマウスを使用し、疼痛モデルにはcomplete Freund's adjuvantの足底注入による炎症性疼痛モデルを用いた。モデルマウス作成後の疼痛急性期から維持期では、S1神経細胞の自発活動が増加し、各細胞間の活動相関性が増加すること、疼痛の改善(作製後28日)に伴ってそれが元の状態まで低下することが分かった。疼痛急性期のS1神経細胞ではN型カルシウムイオンチャネルの発現が増加しており、拮抗薬の脳室内単回投与および徐放製剤を用いたS1局所慢性投与が疼痛改善に有効であった。

  15. Evaluation and Manipulation of Neural Activity using Two-Photon Holographic Microscopy. International journal

    Daisuke Kato, Xiangyu Quan, Yuta Tanisumi, Zhongtian Guo, Mitsuhiro Morita, Tetsuya Takiguchi, Osamu Matoba, Hiroaki Wake

    Journal of visualized experiments : JoVE   Vol. 2022 ( 187 )   2022.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Recent advances in optical bioimaging and optogenetics have enabled the visualization and manipulation of biological phenomena, including cellular activities, in living animals. In the field of neuroscience, detailed neural activity related to brain functions, such as learning and memory, has now been revealed, and it has become feasible to artificially manipulate this activity to express brain functions. However, the conventional evaluation of neural activity by two-photon Ca2+ imaging has the problem of low temporal resolution. In addition, manipulation of neural activity by conventional optogenetics through the optic fiber can only simultaneously regulate the activity of neurons with the same genetic background, making it difficult to control the activity of individual neurons. To solve this issue, we recently developed a microscope with a high spatiotemporal resolution for biological applications by combining optogenetics with digital holographic technology that can modify femtosecond infrared laser beams. Here, we describe protocols for the visualization, evaluation, and manipulation of neural activity, including the preparation of samples and operation of a two-photon holographic microscope (Figure 1). These protocols provide accurate spatiotemporal information on neural activity, which may be useful for elucidating the pathogenesis of neuropsychiatric disorders that lead to abnormalities in neural activity.

    DOI: 10.3791/64205

    Web of Science

    Scopus

    PubMed

  16. Border-associated macrophages transventricularly infiltrate the early embryonic cerebral wall to differentiate into microglia

    Yuki Hattori, Daisuke Kato, Futoshi Murayama, Sota Koike, Yu Naito, Ayano Kawaguchi, Hiroaki Wake, Takaki Miyata

        2022.7

     More details

    Publisher:Cold Spring Harbor Laboratory  

    Summary

    The relationships between microglia and macrophages, especially their lineage segregation outside the yolk sac, have been recently explored, providing a model in which a conversion from macrophages seeds microglia during brain development. However, spatiotemporal evidence to support such microglial seeding and to explain how it occurs has not been obtained. By cell tracking via slice culture, intravital imaging, and Flash tag-mediated labeling, we found that a group of intraventricular macrophages belonging to border-associated macrophages (BAMs), which were abundantly observed along the inner surface of the mouse cerebral wall at embryonic day 12, frequently entered the brain wall. Immunohistochemistry of the tracked cells showed that postinfiltrative BAMs acquired microglial properties while losing a macrophage phenotype. We also found that the intraventricular BAMs were supplied transepithelially from the roof plate. Thus, this study demonstrates that the “roof plate→ventricle→cerebral wall” route is an essential path for microglial colonization into the embryonic mouse brain.

    DOI: 10.1101/2022.07.27.501563

  17. Controlled activation of cortical astrocytes modulates neuropathic pain-like behaviour. International journal

    Ikuko Takeda, Kohei Yoshihara, Dennis L Cheung, Tomoko Kobayashi, Masakazu Agetsuma, Makoto Tsuda, Kei Eto, Schuichi Koizumi, Hiroaki Wake, Andrew J Moorhouse, Junichi Nabekura

    Nature communications   Vol. 13 ( 1 ) page: 4100 - 4100   2022.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Chronic pain is a major public health problem that currently lacks effective treatment options. Here, a method that can modulate chronic pain-like behaviour induced by nerve injury in mice is described. By combining a transient nerve block to inhibit noxious afferent input from injured peripheral nerves, with concurrent activation of astrocytes in the somatosensory cortex (S1) by either low intensity transcranial direct current stimulation (tDCS) or via the chemogenetic DREADD system, we could reverse allodynia-like behaviour previously established by partial sciatic nerve ligation (PSL). Such activation of astrocytes initiated spine plasticity to reduce those synapses formed shortly after PSL. This reversal from allodynia-like behaviour persisted well beyond the active treatment period. Thus, our study demonstrates a robust and potentially translational approach for modulating pain, that capitalizes on the interplay between noxious afferents, sensitized central neuronal circuits, and astrocyte-activation induced synaptic plasticity.

    DOI: 10.1038/s41467-022-31773-8

    Scopus

    PubMed

  18. Lighting up cosmic neuronal networks with transformative in vivo calcium imaging. International journal

    Masanori Murayama, Hiroaki Wake

    Neuroscience research   Vol. 179   page: 1 - 2   2022.6

     More details

  19. Holographic microscope and its biological application. International journal

    Xiangyu Quan, Daisuke Kato, Vincent Daria, Osamu Matoba, Hiroaki Wake

    Neuroscience research   Vol. 179   page: 57 - 64   2022.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Holographic structured illumination combined with optogenetics enables patterned stimulation of neurons and glial cells in an intact living brain. Moreover, in vivo functional imaging of cellular activity with recent advanced microscope technologies allows for visualization of the cellular responses during learning, emotion and cognition. Integrating these techniques can be used to verify the link between cell function and behavior output. However, there are technical limitations to stimulate multiple cells with high spatial and temporal resolution with available techniques of optogenetic stimulation. Here, we summarized a two-photon microscope combined with holographic system to stimulate multiple cells with high spatial and temporal resolution for living mice and their biological application.

    DOI: 10.1016/j.neures.2021.10.012

    Web of Science

    Scopus

    PubMed

  20. A Piezo1/KLF15/IL-6 axis mediates immobilization-induced muscle atrophy International journal

    Yu Hirata, Kazuhiro Nomura, Daisuke Kato, Yoshihisa Tachibana, Takahiro Niikura, Kana Uchiyama, Tetsuya Hosooka, Tomoaki Fukui, Keisuke Oe, Ryosuke Kuroda, Yuji Hara, Takahiro Adachi, Koji Shibasaki, Hiroaki Wake, Wataru Ogawa

    Journal of Clinical Investigation   Vol. 132 ( 10 ) page: 1 - 13   2022.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Society for Clinical Investigation  

    Although immobility is a common cause of muscle atrophy, the mechanism underlying this causality is unclear. We here show that Krüppel-like factor 15 (KLF15) and IL-6 are upregulated in skeletal muscle of limb-immobilized mice and that mice with KLF15 deficiency in skeletal muscle or with systemic IL-6 deficiency are protected from immobility-induced muscle atrophy. A newly developed Ca2+ bioimaging revealed that the cytosolic Ca2+ concentration ([Ca2+]i) of skeletal muscle is reduced to below the basal level by immobilization, which is associated with the downregulation of Piezo1. Acute disruption of Piezo1 in skeletal muscle induced Klf15 and Il6 expression as well as muscle atrophy, which was prevented by antibodies against IL-6. A role for the Piezo1/KLF15/IL-6 axis in immobility-induced muscle atrophy was validated in human samples. Our results thus uncover a paradigm for Ca2+ signaling in that a decrease in [Ca2+]i from the basal level triggers a defined biological event.

    DOI: 10.1172/JCI154611

    Web of Science

    Scopus

    PubMed

  21. Elucidation of the neurological effects of clothianidin exposure at the no-observed-adverse-effect level (NOAEL) using two-photon microscopy &lt;i&gt;in vivo&lt;/i&gt; imaging

    Misaki NISHI, Shouta SUGIO, Tetsushi HIRANO, Daisuke KATO, Hiroaki WAKE, Asuka SHODA, Midori MURATA, Yoshinori IKENAKA, Yoshiaki TABUCHI, Youhei MANTANI, Toshifumi YOKOYAMA, Nobuhiko HOSHI

    Journal of Veterinary Medical Science   Vol. 84 ( 4 ) page: 585 - 592   2022.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japanese Society of Veterinary Science  

    Neonicotinoid pesticides (NNs) cause behavioral abnormalities in mammals, raising concerns about their effects on neural circuit activity. We herein examined the neurological effects of the NN clothianidin (CLO) by in vivo Ca2+ imaging using two-photon microscopy. Mice were fed the no-observed-adverse-effect-level (NOAEL) dose of CLO for 2 weeks and their neuronal activity in the primary somatosensory cortex (S1) was observed weekly for 2 weeks. CLO exposure caused a sustained influx of Ca2+ in neurons in the S1 2/3 layers, indicating hyperactivation of neurons. In addition, microarray gene expression analysis suggested the induction of neuroinflammation and changes in synaptic activity. These results demonstrate that exposure to the NOAEL dose of CLO can overactivate neurons and disrupt neuronal homeostasis.

    DOI: 10.1292/jvms.22-0013

    Web of Science

    Scopus

    PubMed

    CiNii Research

  22. KLF15を介した筋萎縮の分子機構の解明

    平田 悠, 野村 和弘, 加藤 大輔, 橘 吉寿, 新倉 隆宏, 内山 奏, 細岡 哲也, 福井 友章, 大江 啓介, 黒田 良祐, 原 雄二, 安達 貴弘, 柴崎 貢志, 和氣 弘明, 小川 渉

    糖尿病   Vol. 65 ( Suppl.1 ) page: S - 122   2022.4

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

  23. KLF15を介した筋萎縮の分子機構の解明

    平田 悠, 野村 和弘, 加藤 大輔, 橘 吉寿, 新倉 隆宏, 内山 奏, 細岡 哲也, 福井 友章, 大江 啓介, 黒田 良祐, 原 雄二, 安達 貴弘, 柴崎 貢志, 和氣 弘明, 小川 渉

    糖尿病   Vol. 65 ( Suppl.1 ) page: S - 122   2022.4

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

  24. Ca2+ imaging with two-photon microscopy to detect the disruption of brain function in mice administered neonicotinoid insecticides International journal

    Anri Hirai, Shouta Sugio, Collins Nimako, Shouta M. M. Nakayama, Keisuke Kato, Keisuke Takahashi, Koji Arizono, Tetsushi Hirano, Nobuhiko Hoshi, Kazutoshi Fujioka, Kumiko Taira, Mayumi Ishizuka, Hiroaki Wake, Yoshinori Ikenaka

    Scientific Reports   Vol. 12 ( 1 ) page: 5114 - 5114   2022.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media LLC  

    Abstract

    Neonicotinoid pesticides are a class of insecticides that reportedly have harmful effects on bees and dragonflies, causing a reduction in their numbers. Neonicotinoids act as neuroreceptor modulators, and some studies have reported their association with neurodevelopmental disorders. However, the precise effect of neonicotinoids on the central nervous system has not yet been identified. Herein, we conducted in vivo Ca<sup>2+</sup> imaging using a two-photon microscope to detect the abnormal activity of neuronal circuits in the brain after neonicotinoid application. The oral administration of acetamiprid (ACE) (20 mg/kg body weight (BW) in mature mice with a quantity less than the no-observed-adverse-effect level (NOAEL) and a tenth or half of the median lethal dose (LD<sub>50</sub>) of nicotine (0.33 or 1.65 mg/kg BW, respectively), as a typical nicotinic acetylcholine receptor (nAChR) agonist, increased anxiety-like behavior associated with altered activities of the neuronal population in the somatosensory cortex. Furthermore, we detected ACE and its metabolites in the brain, 1 h after ACE administration. The results suggested that in vivo Ca<sup>2+</sup> imaging using a two-photon microscope enabled the highly sensitive detection of neurotoxicant-mediated brain disturbance of nerves.

    DOI: 10.1038/s41598-022-09038-7

    Web of Science

    Scopus

    PubMed

    Other Link: https://www.nature.com/articles/s41598-022-09038-7

  25. Construction of a Multidimensional Brain andBiological Imaging Center for the Creation of a Healthy Society andInternational Collaboration

    Hiroaki Wake

    Proceedings for Annual Meeting of The Japanese Pharmacological Society   Vol. 95 ( 0 ) page: 2-S17-2   2022

     More details

    Language:Japanese   Publisher:Japanese Pharmacological Society  

    <p>Visualization (or &quot;Understanding&quot; or &quot;Researchinto&quot;) of physiological and pathological phenomena has taken a leapforward by recent advanced optical bio imaging technique. Traditionally,physiological phenomena including cell division, cell function, moleculartransport and cell death were detected by biochemical methods. However, thisinformation lacked high resolution spatial and temporal information. Usingrecent optical techniques, both spatial and temporal information can beintegrated across the molecular, cellular and systems levels that allow us tofurther investigate the hierarchical interaction of brain systems thatultimately reveal the pathological mechanism. In this session, we will discussthe current and potential future state of the construction of bio-imagingcenter, and will further discuss by understanding hierarchical physiologicalsystem that leads to the creation of healthy society.</p>

    DOI: 10.1254/jpssuppl.95.0_2-s17-2

    CiNii Research

  26. Microglial function for synapses and vessels in physiological and pathological brain

    Hiroaki Wake

    Proceedings for Annual Meeting of The Japanese Pharmacological Society   Vol. 95 ( 0 ) page: 3-S31-3   2022

     More details

    Language:Japanese   Publisher:Japanese Pharmacological Society  

    <p>Microglia are the sole immune responding cells in the central nervous system. Their role as neuro-immune cells in the pathogenesis of various neurodegenerative and infectious diseases of the brain have been extensively studied. In addition to the pathological function of microglia, recent developments in molecular probes and optical imaging in vivo have revealed that microglia are highly motile cell in the healthy brain, extending and retracting their process that extend from a largely stationary cell soma. We used in vivo two photon microscopy to reveal their physiological and pathological function on synapse and vessels. We particularly showed the functional consequence of microglial contact on synapse and vessels to indicate their role in neurological or psychiatric brain. </p><p>In this session, we will show 1. Microglial regulation of blood brain barrier, 2. Microglial role for cross modal plasticity that indicate their pathological role in schizophrenia. </p><p></p>

    DOI: 10.1254/jpssuppl.95.0_3-s31-3

    CiNii Research

  27. ミクログリアによるシナプス制御

    和氣 弘明, 橋本 明香里

    ファルマシア   Vol. 58 ( 9 ) page: 858 - 861   2022

     More details

    Language:Japanese   Publisher:公益社団法人 日本薬学会  

    アルツハイマー型認知機能障害、自閉スペクトラム症、統合失調症などの多岐にわたる疾患において、神経回路を決定するシナプスの異常を認める。近年の光学技術の発達に伴い、脳の免疫細胞であるミクログリアが、健常時、発達期、病態期の様々な場面で、シナプスと直接の接触を繰り返し、その新生、除去、維持、活動の制御などの多岐にわたる役割を果たすことが解明されてきた。本稿では、ミクログリアとシナプスに関する最新の知見をまとめ、今後の疾患研究への展望について議論する。

    DOI: 10.14894/faruawpsj.58.9_858

    CiNii Research

  28. [Microglial Regulation of Blood Brain Barrier, the Neuro-Immunological Interface].

    Ako Ikegami, Hiroaki Wake

    Brain and nerve = Shinkei kenkyu no shinpo   Vol. 73 ( 8 ) page: 913 - 919   2021.8

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)  

    The central nervous system (CNS) is an immune-privileged area. The blood-brain barrier (BBB) is thought to separate the CNS from any systemic inflammatory states to maintain homeostasis within this specialized, vulnerable organ. However, accumulating studies have challenged this concept by demonstrating systemic inflammatory effects on brain. Moreover, the coronavirus disease pandemic caused by severe acute respiratory syndrome coronavirus 2 in 2019 has rapidly evoked attention toward the BBB as the systemic-CNS immunological interface. In this review, we focus on microglia, the sole immune cells in the CNS, and briefly introduce our new findings regarding microglial BBB regulation in systemic inflammation. With a close eye on associated literature, we carefully rethink the traditional immune system in the CNS and suggest a new possible mechanism of systemic-CNS immune cell interaction, while an understanding of the BBB develops.

    DOI: 10.11477/mf.1416201861

    PubMed

    CiNii Research

  29. [A New Outlook on Mental Disorders: Pathological Dynamics of Glial Cells].

    Yuko Arioka, Daisuke Kato, Hiroaki Wake, Norio Ozaki

    Brain and nerve = Shinkei kenkyu no shinpo   Vol. 73 ( 7 ) page: 787 - 794   2021.7

     More details

    Language:Japanese   Publishing type:Research paper (scientific journal)  

    The pathophysiology of mental disorders remains unknown. This causes many gaps in pathophysiology, diagnosis, and treatment of mental disorders. To close these gaps, a new perspective, which is not bound by the existing diagnostic classifications or pathological hypotheses for mental disorders, is required. Recently, it has been reported that glial cells play active roles in normal brain function and circuit formation, and their disruption results in the onset of mental disorders. Here, we discuss mental disorders from the perspective of glial cell-related pathophysiology, along with our current efforts and research.

    DOI: 10.11477/mf.1416201836

    PubMed

    CiNii Research

  30. 二光子顕微鏡を用いた、ニコチン性アセチルコリン受容体アゴニストによる脳機能障害の検出

    平井 杏梨, 杉尾 翔太, Nimako Collins, 中山 翔太, 加藤 恵介, 高橋 圭介, 有薗 幸司, 平野 哲史, 星 信彦, 藤岡 一俊, 平 久美子, 石塚 真由美, 和氣 弘明, 池中 良徳

    The Journal of Toxicological Sciences   Vol. 46 ( Suppl. ) page: S102 - S103   2021.7

     More details

    Language:English   Publisher:(一社)日本毒性学会  

  31. Myelin plasticity modulates neural circuitry required for learning and behavior. International journal

    Daisuke Kato, Hiroaki Wake

    Neuroscience research   Vol. 167   page: 11 - 16   2021.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Oligodendrocytes, which form the myelin sheaths that insulate axons, regulate conduction velocity. Myelinated axons make up the brain's white matter and contribute to the efficiency of information processing by regulating the timing of neural activity. Traditionally, it has been thought that myelin is a static, inactive insulator around the axon. However, recent studies in humans using magnetic resonance imaging have shown that structural changes in the white matter occur during learning and training, suggesting that 1) white matter change depends on neural activity and 2) activity-dependent changes in white matter are essential for learning and behavior. Furthermore, suppression of oligodendrocytes and their progenitor cells leads to deficits in motor learning and remote fear memory consolidation, suggesting a causal relationship between glial function and the learning process. However, for technical reasons, it remains unclear how myelin-generating glia modulate neural circuitry and what underlying mechanisms they employ to affect learning and behavior. Recent advances in optical and genetic techniques have helped elucidate this mechanism. In this review, we highlight evidence that neural activities regulated by myelin plasticity play a pivotal role in learning and behavior and provide further insight into possible therapeutic targets for treating diseases accompanied by myelin impairment.

    DOI: 10.1016/j.neures.2020.12.005

    Web of Science

    Scopus

    PubMed

  32. Pain induces stable, active microcircuits in the somatosensory cortex that provide a therapeutic target International journal

    Takuya Okada, Daisuke Kato, Yuki Nomura, Norihiko Obata, Xiangyu Quan, Akihito Morinaga, Hajime Yano, Zhongtian Guo, Yuki Aoyama, Yoshihisa Tachibana, Andrew J. Moorhouse, Osamu Matoba, Tetsuya Takiguchi, Satoshi Mizobuchi, Hiroaki Wake

    Science Advances   Vol. 7 ( 12 ) page: eabd8261 - eabd8261   2021.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:American Association for the Advancement of Science (AAAS)  

    Sustained neuropathic pain from injury or inflammation remains a major burden for society. Rodent pain models have informed some cellular mechanisms increasing neuronal excitability within the spinal cord and primary somatosensory cortex (S1), but how activity patterns within these circuits change during pain remains unclear. We have applied multiphoton in vivo imaging and holographic stimulation to examine single S1 neuron activity patterns and connectivity during sustained pain. Following pain induction, there is an increase in synchronized neuronal activity and connectivity within S1, indicating the formation of pain circuits. Artificially increasing neuronal activity and synchrony using DREADDs reduced pain thresholds. The expression of N-type voltage-dependent Ca<sup>2+</sup> channel subunits in S1 was increased after pain induction, and locally blocking these channels reduced both the synchrony and allodynia associated with inflammatory pain. Targeting these S1 pain circuits, via inhibiting N-type Ca<sup>2+</sup> channels or other approaches, may provide ways to reduce inflammatory pain.

    DOI: 10.1126/sciadv.abd8261

    Web of Science

    Scopus

    PubMed

  33. Novel in vivo imaging method to evaluate "Don't eat me" signal of tumor against microglia Reviewed International journal

    Tsuji Takahiro, Wake Hiroaki, Shindo Mariko, Yamazoe Masatoshi, Ajimizu Hitomi, Yasuda Yuto, Funazo Tomoko, Yoshida Hironori, Sakamori Yuichi, Kim Young Hak, Katoh Daisuke, Ozasa Hiroaki, Hirai Toyohiro

    CANCER SCIENCE   Vol. 112   page: 193 - 193   2021.2

     More details

    Publishing type:Research paper (scientific journal)  

    Web of Science

  34. Detection of brain function disruption caused by nicotinic acetylcholine receptor agonists using Ca<sup>2+</sup> imaging with two-photon microscopy

    HIRAI Anri, SUGIO Shouta, NIMAKO Collins, NAKAYAMA Shouta M.M., KATO Keisuke, TAKAHASHI Keisuke, ARIZONO Koji, HIRANO Tetsushi, HOSHI Nobuhiko, FUJIOKA Kazutoshi, TAIRA Kumiko, ISHIZUKA Mayumi, WAKE Hiroaki, IKENAKA Yoshinori

    Annual Meeting of the Japanese Society of Toxicology   Vol. 48.1 ( 0 ) page: P-37E   2021

     More details

    Language:Japanese   Publisher:The Japanese Society of Toxicology  

    <p>Neonicotionoid pesticides are insecticides which have been thought to be one of the causes of Colony Collapse Disorder. They were considered to be less toxic to mammals than insects, but some of the recent studies suggested their association with disruption of higher brain fuction in mammals. Nevertheless, it is still unclear how neonicotionoids affect on the central nervous system. Here, we propose the use of in vivo Ca<sup>2+</sup> imaging with two-photon microscope to detect abnormal activity of neuronal circuits in brain with the application of neonicotionoids.</p><p>In this study, a less than the no-observed-adverse-effect level (NOAEL) of acetamiprid (20 mg/kg bw); and a tenth or half of the median lethal doses of nicotine (0.33 or 1.65 mg/kg bw respectively) were orally administered to mice. They were subjected to elevated plus maze test and Ca<sup>2+</sup> imaging by two-photon microscope in the somatosensory cortex. We further detected acetamiprid and metabolites in brain and blood an hour after the administration.</p><p>Mice exposed to acetamiprid or nicotine exhibited an increase in anxiety-like behavior that associated with the altered activities of the neuronal population in the somatosensory cortex. Although the dose of acetamiprid used in the current study was below the NOAEL, both acetamiprid and nicotine affected the behavior and the neuronal activity in the somatosensory cortex. The results suggested that in vivo Ca<sup>2+</sup> imaging using two-photon microscope enabled highly sensitive detection of brain neurodisruption by neurotoxicants.</p>

    DOI: 10.14869/toxpt.48.1.0_p-37e

    CiNii Research

  35. An alternative method to control and monitor neural activity in 2 photon imaging

    Xiangyu Quan, Daisuke Kato, Hiroaki Wake, Yasuhiro Awatsuji, Osamu Matoba

    Optics InfoBase Conference Papers     2021

     More details

    Publishing type:Research paper (international conference proceedings)  

    Scopus

  36. Maternal immune activation induces sustained changes in fetal microglia motility. Reviewed International journal

    Kana Ozaki, Daisuke Kato, Ako Ikegami, Akari Hashimoto, Shouta Sugio, Zhongtian Guo, Midori Shibushita, Tsuyako Tatematsu, Koichiro Haruwaka, Andrew J Moorhouse, Hideto Yamada, Hiroaki Wake

    Scientific reports   Vol. 10 ( 1 ) page: 21378 - 21378   2020.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Maternal infection or inflammation causes abnormalities in brain development associated with subsequent cognitive impairment and in an increased susceptibility to schizophrenia and autism spectrum disorders. Maternal immune activation (MIA) and increases in serum cytokine levels mediates this association via effects on the fetal brain, and microglia can respond to maternal immune status, but consensus on how microglia may respond is lacking and no-one has yet examined if microglial process motility is impaired. In this study we investigated how MIA induced at two different gestational ages affected microglial properties at different developmental stages. Immune activation in mid-pregnancy increased IL-6 expression in embryonic microglia, but failed to cause any marked changes in morphology either at E18 or postnatally. In contrast MIA, particularly when induced earlier (at E12), caused sustained alterations in the patterns of microglial process motility and behavioral deficits. Our research has identified an important microglial property that is altered by MIA and which may contribute to the underlying pathophysiological mechanisms linking maternal immune status to subsequent risks for cognitive disease.

    DOI: 10.1038/s41598-020-78294-2

    Web of Science

    Scopus

    PubMed

  37. Negative feedback control of neuronal activity by microglia. Reviewed International journal

    Ana Badimon, Hayley J Strasburger, Pinar Ayata, Xinhong Chen, Aditya Nair, Ako Ikegami, Philip Hwang, Andrew T Chan, Steven M Graves, Joseph O Uweru, Carola Ledderose, Munir Gunes Kutlu, Michael A Wheeler, Anat Kahan, Masago Ishikawa, Ying-Chih Wang, Yong-Hwee E Loh, Jean X Jiang, D James Surmeier, Simon C Robson, Wolfgang G Junger, Robert Sebra, Erin S Calipari, Paul J Kenny, Ukpong B Eyo, Marco Colonna, Francisco J Quintana, Hiroaki Wake, Viviana Gradinaru, Anne Schaefer

    Nature   Vol. 586 ( 7829 ) page: 417 - +   2020.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Microglia, the brain's resident macrophages, help to regulate brain function by removing dying neurons, pruning non-functional synapses, and producing ligands that support neuronal survival1. Here we show that microglia are also critical modulators of neuronal activity and associated behavioural responses in mice. Microglia respond to neuronal activation by suppressing neuronal activity, and ablation of microglia amplifies and synchronizes the activity of neurons, leading to seizures. Suppression of neuronal activation by microglia occurs in a highly region-specific fashion and depends on the ability of microglia to sense and catabolize extracellular ATP, which is released upon neuronal activation by neurons and astrocytes. ATP triggers the recruitment of microglial protrusions and is converted by the microglial ATP/ADP hydrolysing ectoenzyme CD39 into AMP; AMP is then converted into adenosine by CD73, which is expressed on microglia as well as other brain cells. Microglial sensing of ATP, the ensuing microglia-dependent production of adenosine, and the adenosine-mediated suppression of neuronal responses via the adenosine receptor A1R are essential for the regulation of neuronal activity and animal behaviour. Our findings suggest that this microglia-driven negative feedback mechanism operates similarly to inhibitory neurons and is essential for protecting the brain from excessive activation in health and disease.

    DOI: 10.1038/s41586-020-2777-8

    Web of Science

    Scopus

    PubMed

  38. 不動化はCa2+シグナルの減弱を通じて筋萎縮を制御する

    平田 悠, 野村 和弘, 新倉 隆宏, 橘 吉寿, 加藤 大輔, 内山 奏, 福井 友章, 大江 啓介, 細岡 哲也, 和氣 弘明, 黒田 良祐, 小川 渉

    糖尿病   Vol. 63 ( Suppl.1 ) page: S - 238   2020.8

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

  39. ニコチン性アセチルコリン受容体アゴニストによるシグナル毒性と2光子顕微鏡を用いた検出手法の開発

    平井 杏梨, 杉尾 翔太, 池中 良徳, Collins Nimako, 中山 翔太, 星 信彦, 和氣 弘明, 石塚 真由美

    The Journal of Toxicological Sciences   Vol. 45 ( Suppl. ) page: S86 - S86   2020.6

     More details

    Language:Japanese   Publisher:(一社)日本毒性学会  

  40. Transient microglial absence assists postmigratory cortical neurons in proper differentiation. Reviewed International journal

    Yuki Hattori, Yu Naito, Yoji Tsugawa, Shigenori Nonaka, Hiroaki Wake, Takashi Nagasawa, Ayano Kawaguchi, Takaki Miyata

    Nature communications   Vol. 11 ( 1 ) page: 1631 - 1631   2020.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    In the developing cortex, postmigratory neurons accumulate in the cortical plate (CP) to properly differentiate consolidating subtype identities. Microglia, despite their extensive surveying activity, temporarily disappear from the midembryonic CP. However, the mechanism and significance of this absence are unknown. Here, we show that microglia bidirectionally migrate via attraction by CXCL12 released from the meninges and subventricular zone and thereby exit the midembryonic CP. Upon nonphysiological excessive exposure to microglia in vivo or in vitro, young postmigratory and in vitro-grown CP neurons showed abnormal differentiation with disturbed expression of the subtype-associated transcription factors and genes implicated in functional neuronal maturation. Notably, this effect is primarily attributed to interleukin 6 and type I interferon secreted by microglia. These results suggest that "sanctuarization" from microglia in the midembryonic CP is required for neurons to appropriately fine-tune the expression of molecules needed for proper differentiation, thus securing the establishment of functional cortical circuit.

    DOI: 10.1038/s41467-020-15409-3

    Web of Science

    Scopus

    PubMed

  41. YAP1 mediates survival of ALK-rearranged lung cancer cells treated with alectinib via pro-apoptotic protein regulation. Reviewed International journal

    Takahiro Tsuji, Hiroaki Ozasa, Wataru Aoki, Shunsuke Aburaya, Tomoko Yamamoto Funazo, Koh Furugaki, Yasushi Yoshimura, Masatoshi Yamazoe, Hitomi Ajimizu, Yuto Yasuda, Takashi Nomizo, Hironori Yoshida, Yuichi Sakamori, Hiroaki Wake, Mitsuyoshi Ueda, Young Hak Kim, Toyohiro Hirai

    Nature communications   Vol. 11 ( 1 ) page: 74 - 74   2020.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Springer Science and Business Media {LLC}  

    Despite the promising clinical efficacy of the second-generation anaplastic lymphoma kinase (ALK) inhibitor alectinib in patients with ALK-rearranged lung cancer, some tumor cells survive and eventually relapse, which may be an obstacle to achieving a cure. Limited information is currently available on the mechanisms underlying the initial survival of tumor cells against alectinib. Using patient-derived cell line models, we herein demonstrate that cancer cells survive a treatment with alectinib by activating Yes-associated protein 1 (YAP1), which mediates the expression of the anti-apoptosis factors Mcl-1 and Bcl-xL, and combinatorial inhibition against both YAP1 and ALK provides a longer tumor remission in ALK-rearranged xenografts when compared with alectinib monotherapy. These results suggest that the inhibition of YAP1 is a candidate for combinatorial therapy with ALK inhibitors to achieve complete remission in patients with ALK-rearranged lung cancer.

    DOI: 10.1038/s41467-019-13771-5

    Web of Science

    PubMed

  42. Motor learning requires myelination to reduce asynchrony and spontaneity in neural activity. Reviewed International journal

    Daisuke Kato, Hiroaki Wake, Philip R Lee, Yoshihisa Tachibana, Riho Ono, Shouta Sugio, Yukio Tsuji, Yasuyo H Tanaka, Yasuhiro R Tanaka, Yoshito Masamizu, Riichiro Hira, Andrew J Moorhouse, Nobuaki Tamamaki, Kazuhiro Ikenaka, Noriyuki Matsukawa, R Douglas Fields, Junichi Nabekura, Masanori Matsuzaki

    Glia   Vol. 68 ( 1 ) page: 193 - 210   2020.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Myelination increases the conduction velocity in long-range axons and is prerequisite for many brain functions. Impaired myelin regulation or impairment of myelin itself is frequently associated with deficits in learning and cognition in neurological and psychiatric disorders. However, it has not been revealed what perturbation of neural activity induced by myelin impairment causes learning deficits. Here, we measured neural activity in the motor cortex during motor learning in transgenic mice with a subtle impairment of their myelin. This deficit in myelin impaired motor learning, and was accompanied by a decrease in the amplitude of movement-related activity and an increase in the frequency of spontaneous activity. Thalamocortical axons showed variability in axonal conduction with a large spread in the timing of postsynaptic cortical responses. Repetitive pairing of forelimb movements with optogenetic stimulation of thalamocortical axon terminals restored motor learning. Thus, myelin regulation helps to maintain the synchrony of cortical spike-time arrivals through long-range axons, facilitating the propagation of the information required for learning. Our results revealed the pathological neuronal circuit activity with impaired myelin and suggest the possibility that pairing of noninvasive brain stimulation with relevant behaviors may ameliorate cognitive and behavioral abnormalities in diseases with impaired myelination.

    DOI: 10.1002/glia.23713

    Web of Science

    Scopus

    PubMed

  43. Signal toxicity caused by nicotinic acetylcholine receptor agonist and development of detection method using two-photon microscope

    HIRAI Anri, SUGIO Shouta, IKENAKA Yoshinori, COLLINS Nimako, NAKAYAMA Shouta M.M, HOSHI Nobuhiko, WAKE Hiroaki, ISHIZUKA Mayumi

    Annual Meeting of the Japanese Society of Toxicology   Vol. 47.1 ( 0 ) page: P-4S - 4S   2020

     More details

    Language:Japanese   Publisher:The Japanese Society of Toxicology  

    <p> By acting on receptors in the brain, some chemicals exhibit "signal toxicity" that disrupts intracellular and intercellular signaling, causing unexpected effects. In particular, in brain function, a complex signal network between neurons and glial cells is formed, and disruption of this network can be a critical endpoint of signal toxicity. However, “disruption of brain function” due to signal toxicity is often not accompanied by cell death and accompanying brain tissue degeneration, and cannot be detected by current neurotoxicity test methods. In this study, we used neonicotinoids (NNs) and nicotine, which are agonists of the nicotinic acetylcholine receptor (nAChR), as model compounds, and detected changes in neuronal activity in the somatosensory cortex using a two-photon microscope. The aim was to detect disruption of brain function associated with signal toxicity.</p><p> C57BL/6N were orally administered ACE at 20 mg/kg (with reference to LOAEL in general pharmacology test of the central nervous system) and their blood was collected over time. It was analyzed quantitatively using LC-MS/MS. Then, we exposed mice to ACE at 20 mg/kg or 7.1 mg/kg (with reference to NOAEL to rats), and their behavior was observed using openfield (OF) test 1 hour after the administration at the former concentration, and using elevated plus maze (EPM) test 1 hour after the administration at the latter concentration. In addition, C57BL/6J were administered ACE at 30 mg/kg (with reference to NOAEL in the subacute toxicity test) and Nicotine, which is the typical agonist of nAChR, at 1.6 mg/kg (with reference to 1/2 LD50), and the neuronal activity in somatosensory cortex was observed over time by using two-photon microscope. </p><p> As a result, ACE absorbed quickly and the Tmax of ACE was around 25 minutes after administration. In comparison, dm-ACE, which is one of its major metabolites, absorbed a bit slower and the Tmax was around 150 minutes. An hour after the administration, the activity of mice decreased, and the anxiety-like behavior increased. In the somatosensory cortex, the firing frequency of nerve cells decreased, and the synchronous firing increased, 1 hour and 2.5 hour after ACE administration. Similarly, the firing frequency tended to decrease, and the synchronous firing increased, 5 minutes and 30 minutes after the administration of Nicotine. This suggest that changes in neuronal activity after administration of ACE are the result of ACE acting as a ligand for nAChR because we can see the same trends between ACE and Nicotine. In summary, it was observed that administration of ACE at a concentration that is originally non-toxic, such as NOAEL, affects the behavior and neural activities. This suggests that cranial nerve activity is responsible for changes in behavior.</p>

    DOI: 10.14869/toxpt.47.1.0_p-4s

    CiNii Research

  44. Dual microglia effects on blood brain barrier permeability induced by systemic inflammation. Reviewed International journal

    Koichiro Haruwaka, Ako Ikegami, Yoshihisa Tachibana, Nobuhiko Ohno, Hiroyuki Konishi, Akari Hashimoto, Mami Matsumoto, Daisuke Kato, Riho Ono, Hiroshi Kiyama, Andrew J Moorhouse, Junichi Nabekura, Hiroaki Wake

    Nature communications   Vol. 10 ( 1 ) page: 5816 - 5816   2019.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Microglia survey brain parenchyma, responding to injury and infections. Microglia also respond to systemic disease, but the role of blood-brain barrier (BBB) integrity in this process remains unclear. Using simultaneous in vivo imaging, we demonstrated that systemic inflammation induces CCR5-dependent migration of brain resident microglia to the cerebral vasculature. Vessel-associated microglia initially maintain BBB integrity via expression of the tight-junction protein Claudin-5 and make physical contact with endothelial cells. During sustained inflammation, microglia phagocytose astrocytic end-feet and impair BBB function. Our results show microglia play a dual role in maintaining BBB integrity with implications for elucidating how systemic immune-activation impacts neural functions.

    DOI: 10.1038/s41467-019-13812-z

    Web of Science

    Scopus

    PubMed

  45. Oral splint ameliorates tic symptoms in patients with tourette syndrome. Reviewed International journal

    Jumpei Murakami, Yoshihisa Tachibana, Shigehisa Akiyama, Takafumi Kato, Aya Taniguchi, Yoshiaki Nakajima, Mao Shimoda, Hiroaki Wake, Yukiko Kano, Masahiko Takada, Atsushi Nambu, Atsushi Yoshida

    Movement disorders : official journal of the Movement Disorder Society   Vol. 34 ( 10 ) page: 1577 - 1578   2019.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1002/mds.27819

    Web of Science

    PubMed

  46. Microglia: Lifelong modulator of neural circuits. Reviewed International journal

    Ako Ikegami, Koichiro Haruwaka, Hiroaki Wake

    Neuropathology : official journal of the Japanese Society of Neuropathology   Vol. 39 ( 3 ) page: 173 - 180   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Microglia, the sole immune cells in the brain, are the key player for synaptic regulation required for our brain function in both developing and adult brain. They have highly motile processes to detect synaptic functions. Recent accumulated studies have unveiled the mechanism underlying synapse detection and pruning / formation by microglia. In this review, we summarize the current knowledge of various microglial machinery recruited in synaptic modulation in the different life stages and contexts.

    DOI: 10.1111/neup.12560

    Web of Science

    PubMed

  47. Activity-Dependent Myelination. Reviewed International journal

    Daisuke Kato, Hiroaki Wake

    Advances in experimental medicine and biology   Vol. 1190   page: 43 - 51   2019

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Oligodendrocyte form myelin around the axons to regulate the conduction velocity. Myelinated axons are composed of white matter to act as cables to connect distinct brain regions. Recent human MRI studies showed that the signal from white matter change in the people with special skills such as taxi driver, piano player, and juggling. The change of the white matter suggested that (1) The plasticity of myelination depends on neuronal activity (activity-dependent myelination) and (2) White matter plasticity is essential for brain functions. In this session, we discussed that how the un-electrical components, oligodendrocytes, and its precursor cells receive the signal from electrically active neurons and differentiate, proliferate, and myelinate the axons to modulate the activity of neuronal circuits, ultimately affect on their behaviors. In this review, we highlight the physiological functions of oligodendrocyte and their neuronal activity-dependent functions and thus show new insight for their contribution to brain functions.

    DOI: 10.1007/978-981-32-9636-7_4

    Web of Science

    PubMed

  48. In Vivo Two-Photon Imaging of Microglial Synapse Contacts. Reviewed International journal

    Daisuke Kato, Ako Ikegami, Hiroshi Horiuchi, Andrew J Moorhouse, Junichi Nabekura, Hiroaki Wake

    Methods in molecular biology (Clifton, N.J.)   Vol. 2034   page: 281 - 286   2019

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Microglia are traditionally known as immune sentinels of the brain and as key player in the pathogenesis of neurodegenerative diseases such as Alzheimer's disease, Parkinson disease, or amyotrophic lateral sclerosis. Recently, they were also identified as synaptic organizer, promoting formation and maturation of synapses as well as modifying synaptic activity. Interestingly, microglia-mediated synaptic pruning and microglia-mediated changes in synaptic plasticity were observed both during brain development and in neurodegenerative diseases, stressing the key role of microglia-synapse interaction in these processes. Here we descried a technique for noninvasive in vivo monitoring of microglia-synapse interactions by means of two-photon microscopy.

    DOI: 10.1007/978-1-4939-9658-2_20

    Web of Science

    PubMed

  49. Physiological Implications of Microglia-Synapse Interactions. Reviewed International journal

    Hiroaki Wake, Hiroshi Horiuchi, Daisuke Kato, Andrew J Moorhouse, Junichi Nabekura

    Methods in molecular biology (Clifton, N.J.)   Vol. 2034   page: 69 - 80   2019

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Microglia are the sole immune responding cells in the central nervous system. Their role as neuroimmune cells in the pathogenesis of various neurodegenerative and infectious diseases of the brain have been extensively studied. Upon brain disease and infection, they adopt an activated phenotype associated with the release of cytokines and neurotrophic factors and resulting in neuroprotective or neurotoxic outcomes. However, microglia are resident also in the healthy or physiological brain, but much less is known about their role(s) in the healthy brain, partly due to technical limitations regarding investigation of these highly reactive cells in the intact brain. Recent developments in molecular probes and in vivo optical imaging techniques has now helped to characterize microglia in the physiological or healthy brain. In vivo two-photon imaging of fluorescently labeled microglia have revealed that they are highly motile cells in the healthy brain, extending and retracting their processes that extend from a largely stationary cell soma. In this chapter, we briefly summarize some of the physiological functions of microglia in the uninjured brain, with a focus on interactions they have with synapses.

    DOI: 10.1007/978-1-4939-9658-2_6

    Web of Science

    PubMed

  50. Erratum: Regulation of myelin structure and conduction velocity by perinodal astrocytes (Proceedings of the National Academy of Sciences of the United States of America (2018) 115 (11832–11837) DOI: 10.1073/pnas.1811013115)

    Dutta, D.J., Woo, D.H., Lee, P.R., Pajevic, S., Bukalo, O., Huffman, W.C., Wake, H., Basser, P.J., SheikhBahaei, S., Lazarevic, V., Smith, J.C., Douglas Fields, R.

    Proceedings of the National Academy of Sciences of the United States of America   Vol. 116 ( 25 ) page: 12574 - 12574   2019

     More details

    Publishing type:Research paper (scientific journal)  

    DOI: 10.1073/pnas.1908361116

    Web of Science

    Scopus

  51. Regulation of myelin structure and conduction velocity by perinodal astrocytes. Reviewed International journal

    Dipankar J Dutta, Dong Ho Woo, Philip R Lee, Sinisa Pajevic, Olena Bukalo, William C Huffman, Hiroaki Wake, Peter J Basser, Shahriar SheikhBahaei, Vanja Lazarevic, Jeffrey C Smith, R Douglas Fields

    Proceedings of the National Academy of Sciences of the United States of America   Vol. 115 ( 46 ) page: 11832 - 11837   2018.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The speed of impulse transmission is critical for optimal neural circuit function, but it is unclear how the appropriate conduction velocity is established in individual axons. The velocity of impulse transmission is influenced by the thickness of the myelin sheath and the morphology of electrogenic nodes of Ranvier along axons. Here we show that myelin thickness and nodal gap length are reversibly altered by astrocytes, glial cells that contact nodes of Ranvier. Thrombin-dependent proteolysis of a cell adhesion molecule that attaches myelin to the axon (neurofascin 155) is inhibited by vesicular release of thrombin protease inhibitors from perinodal astrocytes. Transgenic mice expressing a dominant-negative fragment of VAMP2 in astrocytes, to reduce exocytosis by 50%, exhibited detachment of adjacent paranodal loops of myelin from the axon, increased nodal gap length, and thinning of the myelin sheath in the optic nerve. These morphological changes alter the passive cable properties of axons to reduce conduction velocity and spike-time arrival in the CNS in parallel with a decrease in visual acuity. All effects were reversed by the thrombin inhibitor Fondaparinux. Similar results were obtained by viral transfection of tetanus toxin into astrocytes of rat corpus callosum. Previously, it was unknown how the myelin sheath could be thinned and the functions of perinodal astrocytes were not well understood. These findings describe a form of nervous system plasticity in which myelin structure and conduction velocity are adjusted by astrocytes. The thrombin-dependent cleavage of neurofascin 155 may also have relevance to myelin disruption and repair.

    DOI: 10.1073/pnas.1811013115

    Web of Science

    PubMed

  52. Three-dimensional stimulation and imaging-based functional optical microscopy of biological cells. Reviewed International journal

    Xiangyu Quan, Manoj Kumar, Osamu Matoba, Yasuhiro Awatsuji, Yoshio Hayasaki, Satoshi Hasegawa, Hiroaki Wake

    Optics letters   Vol. 43 ( 21 ) page: 5447 - 5450   2018.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    A new type of functional optical microscope system called three-dimensional (3D) stimulation and imaging-based functional optical microscopy (SIFOM) is proposed, to the best of our knowledge. SIFOM can precisely stimulate user-defined targeted biological cells and can simultaneously record the volumetric fluorescence distribution in a single acquisition. Precise and simultaneous stimulation of fluorescent-labeled biological cells is achieved by multiple 3D spots generated by digital holograms displayed on a phase-mode spatial light modulator. Single-shot 3D acquisition of the fluorescence distribution is accomplished by common-path off-axis incoherent digital holographic microscopy in which a diffraction grating with a focusing lens is displayed on another phase-mode spatial light modulator. The effectiveness of the proposed functional microscope system was verified in experiments using fluorescent microbeads and human lung cancer cells located at various defocused positions. The system can be used for manipulating the states of cells in optogenetics.

    DOI: 10.1364/OL.43.005447

    Web of Science

    PubMed

  53. Microglia mediate non-cell-autonomous cell death of retinal ganglion cells. Reviewed International journal

    Akiko Takeda, Youichi Shinozaki, Kenji Kashiwagi, Nobuhiko Ohno, Kei Eto, Hiroaki Wake, Junichi Nabekura, Schuichi Koizumi

    Glia   Vol. 66 ( 11 ) page: 2366 - 2384   2018.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Excitotoxicity is well known in the neuronal death in the brain and is also linked to neuronal damages in the retina. Recent accumulating evidence show that microglia greatly affect excitotoxicity in the brain, but their roles in retina have received only limited attention. Here, we report that retinal excitotoxicity is mediated by microglia. To this end, we employed three discrete methods, that is, pharmacological inhibition of microglia by minocycline, pharmacological ablation by an antagonist for colony stimulating factor 1 receptor (PLX5622), and genetic ablation of microglia using Iba1-tTA::DTAtetO/tetO mice. Intravitreal injection of NMDA increased the number of apoptotic retinal ganglion cells (RGCs) followed by reduction in the number of RGCs. Although microglia did not respond to NMDA directly, they became reactive earlier than RGC damages. Inhibition or ablation of microglia protected RGCs against NMDA. We found up-regulation of proinflammatory cytokine genes including Il1b, Il6 and Tnfa, among which Tnfa was selectively blocked by minocycline. PLX5622 also suppressed Tnfa expression. Tumor necrosis factor α (TNFα) signals were restricted in microglia at very early followed by spreading into other cell types. TNFα up-regulation in microglia and other cells were significantly attenuated by minocycline and PLX5622, suggesting a central role of microglia for TNFα induction. Both inhibition of TNFα and knockdown of TNF receptor type 1 by siRNA protected RGCs against NMDA. Taken together, our data demonstrate that a phenotypic change of microglia into a neurotoxic one is a critical event for the NMDA-induced degeneration of RGCs, suggesting an importance of non-cell-autonomous mechanism in the retinal neuronal excitotoxicity.

    DOI: 10.1002/glia.23475

    Web of Science

    PubMed

  54. Microglia enhance synapse activity to promote local network synchronization Reviewed International journal

    Akiyoshi, R., Wake, H., Kato, D., Horiuchi, H., Ono, R., Ikegami, A., Haruwaka, K., Omori, T., Tachibana, Y., Moorhouse, A.J., Nabekura, J.

    eNeuro   Vol. 5 ( 5 )   2018.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    DOI: 10.1523/ENEURO.0088-18.2018

    Web of Science

    Scopus

    PubMed

  55. Cortical astrocytes prime the induction of spine plasticity and mirror image pain. Reviewed International journal

    Tatsuya Ishikawa, Kei Eto, Sun Kwang Kim, Hiroaki Wake, Ikuko Takeda, Hiroshi Horiuchi, Andrew J Moorhouse, Hitoshi Ishibashi, Junichi Nabekura

    Pain   Vol. 159 ( 8 ) page: 1592 - 1606   2018.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Peripheral nerve injury causes maladaptive plasticity in the central nervous system and induces chronic pain. In addition to the injured limb, abnormal pain sensation can appear in the limb contralateral to the injury, called mirror image pain. Because synaptic remodeling in the primary somatosensory cortex (S1) has critical roles in the induction of chronic pain, cortical reorganization in the S1 ipsilateral to the injured limb may also accompany mirror image pain. To elucidate this, we conducted in vivo 2-photon calcium imaging of neuron and astrocyte activity in the ipsilateral S1 after a peripheral nerve injury. We found that cross-callosal inputs enhanced the activity of both S1 astrocytes and inhibitory neurons, whereas activity of excitatory neurons decreased. When local inhibitory circuits were blocked, astrocyte-dependent spine plasticity and allodynia were revealed. Thus, we propose that cortical astrocytes prime the induction of spine plasticity and mirror image pain after peripheral nerve injury. Moreover, this result suggests that cortical synaptic rewiring could be sufficient to cause allodynia on the uninjured periphery.

    DOI: 10.1097/j.pain.0000000000001248

    Web of Science

    PubMed

  56. Activity-dependent functions of non-electrical glial cells. Reviewed International journal

    Daisuke Kato, Kei Eto, Junichi Nabekura, Hiroaki Wake

    Journal of biochemistry   Vol. 163 ( 6 ) page: 457 - 464   2018.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Electrical activity is essential for brain function. However, neurons, the electrically active cells, are less numerous than the non-electrical glial cells in the central nervous system. The non-electrical components modify the function of neural circuits, depending on the electrical neuronal activity, by wrapping synapses, myelinating axons and phagocytozing the neuronal components. Moreover, recent evidence has suggested that they contribute to neurological and psychiatric disease by regulating neuronal circuits, ultimately affecting their behaviour. In this review, we highlight the physiological functions of glial cells, particularly the electrical activity-dependent processes, to provide further insight into their role in brain function.

    DOI: 10.1093/jb/mvy023

    Web of Science

    Scopus

    PubMed

  57. Three-dimensional stimulation and imaging-based functional optical microscopy of biological cells Reviewed International journal

    Xiangyu Quan, Manoj Kumar, MATOBA Osamu, Yasuhiro Awatsuji, Yoshio Hayasaki, Satoshi Hasegawa, WAKE Hiroki

    Optics Letters   Vol. Vol. 43 ( No. 21 ) page: pp. 5447 - 5480   2018

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

  58. High temporal and spatial pattern stimulation to manipulate brain function

    Wake, H; Haruwaka, K; Zen, K; Matoba, O

    BIOMEDICAL IMAGING AND SENSING CONFERENCE   Vol. 10711   2018

  59. Reduced Mastication Impairs Memory Function Reviewed International journal

    Y. Fukushima-Nakayama, Takehito Ono, M. Hayashi, M. Inoue, H. Wake, Takashi Ono, T. Nakashima

    JOURNAL OF DENTAL RESEARCH   Vol. 96 ( 9 ) page: 1058 - 1066   2017.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SAGE PUBLICATIONS INC  

    Mastication is an indispensable oral function related to physical, mental, and social health throughout life. The elderly tend to have a masticatory dysfunction due to tooth loss and fragility in the masticatory muscles with aging, potentially resulting in impaired cognitive function. Masticatory stimulation has influence on the development of the central nervous system (CNS) as well as the growth of maxillofacial tissue in children. Although the relationship between mastication and cognitive function is potentially important in the growth period, the cellular and molecular mechanisms have not been sufficiently elucidated. Here, we show that the reduced mastication resulted in impaired spatial memory and learning function owing to the morphological change and decreased activity in the hippocampus. We used an in vivo model for reduced masticatory stimuli, in which juvenile mice were fed with powder diet and found that masticatory stimulation during the growth period positively regulated long-term spatial memory to promote cognitive function. The functional linkage between mastication and brain was validated by the decrease in neurons, neurogenesis, neuronal activity, and brain-derived neurotrophic factor (BDNF) expression in the hippocampus. These findings taken together provide in vivo evidence for a functional linkage between mastication and cognitive function in the growth period, suggesting a need for novel therapeutic strategies in masticatory function-related cognitive dysfunction.

    DOI: 10.1177/0022034517708771

    Web of Science

    PubMed

▼display all

MISC 12

  1. 不動化はPiezo1/KLF15経路を介して筋萎縮を促進する

    平田 悠, 野村 和弘, 加藤 大輔, 橘 吉寿, 内山 奏, 細岡 哲也, 原 雄二, 安達 貴弘, 柴崎 貢志, 和氣 弘明, 小川 渉

    糖尿病   Vol. 65 ( Suppl.1 ) page: S - 185   2022.4

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

  2. 不動化はPiezo1/KLF15経路を介して筋萎縮を促進する

    平田 悠, 野村 和弘, 加藤 大輔, 橘 吉寿, 内山 奏, 細岡 哲也, 原 雄二, 安達 貴弘, 柴崎 貢志, 和氣 弘明, 小川 渉

    糖尿病   Vol. 65 ( Suppl.1 ) page: S - 185   2022.4

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

  3. 不動化はPiezo1/KLF15経路を介して筋萎縮を促進する

    平田 悠, 野村 和弘, 加藤 大輔, 橘 吉寿, 内山 奏, 細岡 哲也, 原 雄二, 安達 貴弘, 柴崎 貢志, 和氣 弘明, 小川 渉

    肥満研究   Vol. 27 ( Suppl. ) page: 328 - 328   2022.3

     More details

    Language:Japanese   Publisher:(一社)日本肥満学会  

  4. 骨格筋生体イメージングを活用した不動化性筋萎縮の発症メカニズムの解析

    平田 悠, 野村 和弘, 加藤 大輔, 橘 吉寿, 内山 奏, 細岡 哲也, 和氣 弘明, 安達 貴弘, 小川 渉

    糖尿病   Vol. 64 ( Suppl.1 ) page: II - 1   2021.5

     More details

    Language:Japanese   Publisher:(一社)日本糖尿病学会  

  5. The disruption of brain function in mice administered with nicotinic acetylcholine receptor agonists~The development of the detection method using two-photon microscopy~

    平井杏梨, 杉尾翔太, NIMAKO Collins, 中山翔太, 加藤恵介, 高橋圭介, 有薗幸司, 平野哲史, 星信彦, 石塚真由美, 和氣弘明, 池中良徳

    環境化学討論会要旨集(CD-ROM)   Vol. 29th   2021

     More details

  6. 不動化はCa2+シグナルの減弱を通じて筋萎縮を制御する

    平田 悠, 野村 和弘, 新倉 隆宏, 橘 吉寿, 加藤 大輔, 内山 奏, 福井 友章, 大江 啓介, 細岡 哲也, 和氣 弘明, 黒田 良祐, 小川 渉

    日本内分泌学会雑誌   Vol. 96 ( 1 ) page: 248 - 248   2020.8

     More details

    Language:Japanese   Publisher:(一社)日本内分泌学会  

    J-GLOBAL

  7. オリゴデンドロサイトおよびその前駆細胞の生体内カルシウムイメージング

    尾野里穂, 加藤大輔, 杉尾翔太, 橘吉寿, 和氣弘明

    日本生理学雑誌(Web)   Vol. 82 ( 1 )   2020

     More details

  8. 行動毒性試験および二光子イメージングによるアセタミプリドの神経毒性評価

    平井杏梨, 杉尾翔太, 池中良徳, COLLINS Nimako, 中山翔太, 星信彦, 和氣弘明, 石塚真由美

    日本獣医学会学術集会講演要旨集   Vol. 163rd   2020

     More details

  9. 妊娠中の母体炎症による児ミクログリアへの影響

    尾崎可奈, 春若航一郎, 橘吉寿, 加藤大輔, 和氣弘明

    日本生理学雑誌(Web)   Vol. 82 ( 1 )   2020

     More details

  10. Real-time visualization of brain metastasis in vivo Reviewed International journal

    Takahiro Tsuji, Hiroaki Wake, Hiroaki Ozasa, Koichiro Haruwaka, Hitomi Ajimizu, Yuto Yasuda, Yuichi Sakamori, Takashi Nomizo, Young Hak Kim, Toyohiro Hirai

    AACR Annual Meeting 2019   Vol. April 1, 2019   2019.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)  

  11. Simple intensity equalization methods in SLM generated multispots

    Xiangyu Quan, Manoj Kumar, Osamu Matoba, Yasuhiro Awatsuji, Hiroaki Wake

    Proceedings of SPIE - The International Society for Optical Engineering   Vol. 11140   2019

     More details

    © 2019 SPIE. Multispots in three-dimensions generated by phase mode spatial light modulators (SLMs) are very useful in laser processing or light induced biological treatments, such as optogenetics. So far, the intensity of the focus spots varies depending to the distance from optical axis. In order to equalize intensities in all generated spots, simple feedback method was applied. Further, look up table was created by dividing imaging area in sectors. Experiment results show improvement in the random spots at some degree, however, further analyzation is required.

    Scopus

  12. イオンイメージセンサによる脳内pHイメージング

    堀内浩, 石田順子, 中村友亮, 稲田浩之, 揚妻正和, 和氣弘明, 澤田和明, 鍋倉淳一

    日本生理学雑誌(Web)   Vol. 80 ( 1 ) page: WEB ONLY   2018.2

     More details

    Language:Japanese  

    J-GLOBAL

▼display all

Presentations 87

  1. 2光子励起を用いたホログラフィック光刺激の現状:生体試料への適用を目指して

    SUGIO SHOUTA, TACHIBANA YOSHIHISA, WAKE HIROAKI

    CREST第2回シンポジウム  2018.12 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:西尾  

  2. 2光子顕微鏡による生体イメージング

    Wake Hiroaki

    京都大学呼吸器内科  2017.9 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:京都  

  3. 2光子顕微鏡による生体イメージング

    Wake Hiroaki

    京都大学セミナー  2017.8 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:京都  

  4. 2光子励起を用いたホログラフィック光刺激法の開発:現状と展望

    SUGIO SHOUTA, TACHIBANA YOSHIHISA, WAKE HIROAKI

    シグナル伝達医学研究展開センター若手道場  2019.1 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:淡路  

  5. A New Type of Microscopy for Light Stimulation and 3D Imaging; Invited International conference

    Xiangyu Quan, Manoj Kumar, MATOBA Osamu, Yasuhiro Awatsuji, Yoshio Hayasaki, Satoshi Hasegawa, WAKE Hiroaki, Mitsuhiro Morita

    International Workshop on Holography and Related Technologies 2018 (IWH2018)  2018.11  IWH2018

     More details

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:Suzhou; China;  

  6. Abnormal Behavior and Malformation of Microglia in Schizophrenic Mice International conference

    Ako Ikegami, Koichiro Haruwaka, Wake Hiroaki

    第2回神戸大学・ワシントン大学・オスロ大学国際合同シンポジウム  2018.3 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:ハワイ, 米国  

  7. Activity dependent myelin regulation in information processing

    Wake Hiroaki

    第115回IIISセミナー  2017.8 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:茨城  

  8. Ca2+ imaging of oligodendrocyte and oligodendrocyte precursor cell in vivo

    Riho Ono, Shouta Sugio, Yoshihisa Tachibana, Hiroaki Wake

    新学術領域研究 スクラップ&ビルドによる脳機能の動的制御 第3回領域会議  2018.11 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:大阪  

  9. Change of lipid profile in myelin associated with motor learning affect on neuronal activity pattern in primary motor cortex.

    Kazuki Nishida, Yoshihisa Tachibana, Shumpei Sato, Fumiyoshi Yamazaki, Mitsutoshi Setou, Hiroaki Wake

    第23 回グリア研究会  2018.12 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:名古屋  

  10. Ca2+ imaging of oligodendrocyte and oligodendrocyte precursor cell in vivo

    Shouta Sugio, Riho Ono, Yoshihisa Tachibana, Hiroaki Wake

    次世代脳プロジェクト 冬のシンポジウム  2018.12 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:東京  

  11. Fetal microglia changes by maternal immune activation

    Kana Ozaki, Wake Hiroaki, Hideto Yamada

    第70回日本産科婦人科学会学術講演会  2018.5 

     More details

    Language:English   Presentation type:Symposium, workshop panel (public)  

    Venue:仙台  

  12. Lipid composition of frontal white matter is altered with motor learning.

    Yukio Tsuji, Tachibana Yoshihisa, Fumiyoshi Yamazaki, Daisuke Kato, Shinohara Masakazu, Miyamoto Akiko, Ikuko Yao, Toda Tatsushi, Mitsutoshi Setou, Wake Hiroaki

    第40回日本神経科学大会  2017.7 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:千葉  

  13. In vivo tracing of single neuron activity with Ca2+ imaging of primary somatosensory cortex in mouse models of postoperative pain and inflammatory pain International conference

    岡田 卓也, 橘 吉寿, NOMURA YUKI, OBATA NORIHIKO, MIZOBUCHI SATOSHI, WAKE HIROAKI

    Cold Spring Harbor Asia Confference  2018.9 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:淡路  

  14. In vivo tracing of single neuron activity with Ca2+ imaging of primary somatosensory cortex in mouse models of postoperative pain and inflammatory pain International conference

    Takuya Okada, Yoshihisa Tachibana, Yuki Nomura, Norihiko Obata, Satoshi Mizobuchi, Hiroaki Wake

    Neuroscience 2018  2018.11 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:San Diego, CA  

  15. High Temporal and Spatial Pattern Stimulation to Manipulate Brain Function Invited International conference

    Hiroaki Wake

    International Workshop on Bioimaging2019  2019.2 

     More details

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:宇都宮  

  16. High Temporal and Spatial Pattern Stimulation to Manipulate Brain Function International conference

    Hiroaki Wake

    ABiS International Symposium  2019.2 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:岡崎  

  17. High Temporal and Spatial Pattern Stimulation to Manipulate Brain Function International conference

    Hiroaki Wake, Koichiro Haruwaka, Xiangyu Quan, Osamu Matoba

    OPIC2018  2018.4 

     More details

    Language:English   Presentation type:Oral presentation (general)  

    Venue:横浜  

  18. High Temporal and Spatial Pattern Stimulation to Manipulate Brain Function International conference

    WAKE Hiroaki, Koichiro Haruwaka, Xiangyu Quan, MATOBA Osamu

    BISC  2018.4  OPIC2018

     More details

    Language:English   Presentation type:Oral presentation (general)  

    Venue:神奈川、横浜  

  19. Microglia and Synapses Invited International conference

    Hiroaki Wake

    ICN2018 世界神経病理学会  2018.9 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:東京  

  20. Microglia in health and disease -lesson from schizophreniamodel mice Invited

    WAKE HIROAKI

    第41回日本神経科学大会  2018.7 

     More details

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:神戸  

  21. Microglia in physiological brain—Focusing on microglia—synapse interactions Invited International conference

    Hiroaki Wake

    Cold Spring Harbor Asia Conferences  2018.12 

     More details

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:蘇州  

  22. Microglia in Health and Disease.- Interaction with synapses- Invited

    Hiroaki Wake

    日本生化学大会  2018.9 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:京都  

  23. Microglia sense systemic immune status to modify activity of neuronal circuit

    Koichiro Haruwaka, Wake Hiroaki, Junichi Nabekura

    Life Science Retreat 2017  2017.10 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:山梨  

  24. Microglia sense systemic immune status to modify activity of neuronal circuit; 免疫状態依存的なミクログリアによる神経回路活動の制御

    Koichiro Haruwaka, Wake Hiroaki, Junichi Nabekura

    第40回日本神経科学大会  2017.7 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:千葉  

  25. Microglial contribute to dendritic spine formation in postnatal mice somatosensory cortex Invited

    Miyamoto Akiko, Wake Hiroaki, Ayako Ishikawa, Hideji Murakoshi, Kei Eto, Yumiko Yoshimura, Junichi Nabekura

    日本神経化学学会  2017.9 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:仙台  

  26. Myelination for information processing

    Wake Hiroaki

    第90回日本神経化学会大会  2017.9 

     More details

    Language:English   Presentation type:Oral presentation (general)  

    Venue:仙台  

  27. Multimodal Digital Holographic Microscopy Invited International conference

    Xiangyu Quan, Manoj Kumar, MATOBA Osamu, Yasuhiro Awatsuji, Yoshio Hayasaki, Satoshi Hasegawa, WAKE Hiroaki, Mitsuhiro Morita

    The 8th Japana-Korea workshop on digital holography and information photonics (DHIP 2018)  2018.11  DHIP2018

     More details

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:Osaka; Japan  

  28. Neurochemistry of neuron-glia interaction Invited International conference

    Hiroaki Wake

    WFSBP2018 The ISN-JSN  2018.9 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:神戸  

  29. oligodendrocyte and myelin remodeking in information processing

    Wake Hiroaki

    第2回「スクラップビルド」領域会議  2017.8 

     More details

    Language:English   Presentation type:Oral presentation (general)  

    Venue:山梨  

  30. Physiclogocal function of microglia and their role for disease formation

    WAKE HIROAKI

    武田薬品社内シンポジウム  2018.12 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:藤沢  

  31. Physiology of microglia -New Insights- Invited International conference

    Wake Hiroaki

    Physiology of microglia XXIII World Congres of Neurology  2017.9 

     More details

    Language:English   Presentation type:Symposium, workshop panel (nominated)  

    Venue:京都  

  32. Physiology of microglia Invited

    Wake Hiroaki

    九州大学歯学部  2017.9 

     More details

    Language:English   Presentation type:Oral presentation (invited, special)  

    Venue:福岡  

  33. Physiologiacl function of microglia and their effect on neuronal circuits International conference

    Hiroaki Wake

    第9回アジアオセアニア生理学会(神戸市)  2019.3 

     More details

    Language:English   Presentation type:Symposium, workshop panel (public)  

    Venue:神戸  

  34. The response to whisker stimulation in the visual cortex of monocular deprived mice in vivo International conference

    橋本 明香里, MIYAMOTO AKIKO, TACHIBANA YOSHIHISA, 春若航一路, WAKE HIROAKI

    9th FAOPS  2019.3 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:神戸  

  35. The scrap and build of oligodendrocyte function and myelin components

    WAKE HIROAKI

    スクラップ&ビルド領域会議  2018.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:大阪  

  36. Two-photon in vivo imaging of oligodendrocyte Ca2+ activity in mice International conference

    Riho Ono, Shouta Sugio, Yoshihisa Tachibana, Hiroaki Wake

    Young glia  2018.10 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:シュパイヤー  

  37. Visualization and manipulation of glial cell functions in vivo International conference

    Hiroaki Wake

    glial heterogeneity SPP1757  2018.10 

     More details

    Language:English   Presentation type:Oral presentation (general)  

    Venue:シュパイヤー  

  38. 光で照らし出すグリアの新機能および脳機能表出

    Wake Hiroaki

    Glial assembly, The 5th Summer Workshop  2017.6 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:新潟  

  39. 光で照らし出すグリアの新機能および脳機能への寄与 Invited

    Wake Hiroaki

    (株)ファイザー 第12回緑内障若手研究者の会(東京)  2017.10 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:東京  

  40. 中枢神経系免疫細胞ミクログリアの新規生理機能とその病態への関与

    WAKE HIROAKI

    日本薬学会北陸支部特別講演会  2018.12 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:金沢  

  41. ミクログリアの生理機能とその破綻による疾患について

    WAKE HIROAKI

    東京女子医科大セミナー  2018.10 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:東京  

  42. ミクログリアの生理・病態を踏まえた認知症治療戦略 Invited

    WAKE HIROAKI

    第37回日本認知症学会学術集会  2018.10 

     More details

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:札幌  

  43. ミクログリアの新規生理機能の可視化

    WAKE HIROAKI

    北海道大学医学部免疫代謝内科学セミナー  2019.2 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:札幌  

  44. ミクログリアの新規生理機能とその破綻による病態の表出

    WAKE HIROAKI

    第11回先端脳科学セミナー  2019.2 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:中央  

  45. ミクログリアによる神経機能修飾

    WAKE HIROAKI

    Neurovascular Unit研究会2019  2019.1 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:東京  

  46. ホログラフィック光照射による蛍光励起の強度均一化"

    小管啓仁, Xiangyu Quan, MATOBA Osamu, 早崎芳夫, 粟辻安浩, WAKE Hiroaki

    日本光学会年次学術講演会(Optics & Photonics Japan 2018)  2018.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:筑波大学東京キャンパス文京校舎,東京  

  47. グリア細胞の生理機能と神経回路機能

    WAKE HIROAKI

    第48回小児神経セミナー  2018.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:大阪  

  48. グリアの生理機能とその破綻による精神疾患の可能性

    WAKE HIROAKI

    動的恒常性研究会  2018.6 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:東京  

  49. オリゴデンドロサイトおよびその前駆細胞のin vivo カルシウムイメージング

    尾野 里穂, SUGIO SHOUTA, TACHIBANA YOSHIHISA, WAKE HIROAKI

    シグナル伝達医学研究展開センター若手道場  2019.1 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:淡路  

  50. What we see in microglia about schizophrenia

    池上暁湖, 春若航一路, TACHIBANA YOSHIHISA, WAKE HIROAKI

    第5回先進イメージング医学研究会  2018.8 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:京都  

  51. 光で迫る脳免疫細胞の機能

    Wake Hiroaki

    脳科学の達人2017  2017.6 

     More details

    Language:Japanese   Presentation type:Symposium, workshop panel (public)  

    Venue:東京  

  52. Microglia sense systemic immune activation in autoimmune disease model.

    Koichiro Haruwaka, Junichi Nabekura, Wake Hiroaki

    次世代脳冬のシンポジウム  2017.12 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:東京  

  53. 自己免疫疾患におけるミクログリアの変化

    春若 航一路, 鍋倉 淳一, Wake Hiroaki

    第111回近畿生理学談話会  2017.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:兵庫  

  54. 脳科学について

    WAKE HIROAKI

    santec社内講演会  2018.10 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:小牧  

  55. 統合失調症モデルマウスにおけるミクログリア動態・形態の変容

    Ako Ikegami, Koichiro Haruwaka, Wake Hiroaki

    新学術領域 S&B第2回班会議  2017.8 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:山梨  

  56. 統合失調症モデルマウスにおけるミクログリア動態・形態の変容

    Ako Ikegami, Koichiro Haruwaka, Wake Hiroaki

    次世代脳冬のシンポジウム  2017.12 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:東京  

  57. 統合失調症モデルマウスにおけるミクログリア動態・形態の変容

    Ako Ikegami, Koichiro Haruwaka, Wake Hiroaki

    第7回ニューロサイエンス研究会  2017.12 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:兵庫  

  58. 統合失調症モデルマウスにおけるミクログリア動態・形態の変容

    Ako Ikegami, Koichiro Haruwaka, Wake Hiroaki

    第110回近畿生理学談話会  2017.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:兵庫  

  59. Optical control of neuron and glial cells by the 3D multi-points stimulation

    春若 航一路, 全 香玉, 森田 光洋, Miyamoto Akiko, 的場 修, Wake Hiroaki

    CREST「光の特性を活用した生命機能の時空間制御技術の開発と応用」研究領域 第2回領域会議  2018.1 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:東京  

  60. 生体イメージング最前線

    WAKE HIROAKI

    小野薬品工業「脳移転治療について考える会」  2018.9 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:京都  

  61. 生体イメージングが明らかにする脳-免疫系の相互作用

    春若 航一路, 鍋倉 淳一, Wake Hiroaki

    岡崎信用金庫先端奨学金制度 成果発表会  2018.2 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:愛知  

  62. 生体 in Vivoイメージング

    WAKE HIROAKI

    第2回ニコンイメージングフォーラム  2019.2 

     More details

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:東京  

  63. 大脳皮質感覚野の生体イメージングによる疼痛発症機構解明へのアプローチ

    岡田 卓也, TACHIBANA YOSHIHISA, NOMURA YUKI, OBATA NORIHIKO, MIZOBUCHI SATOSHI, WAKE HIROAKI

    第111回 近畿生理学談話会  2018.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:和歌山  

  64. 単眼遮蔽による視覚野のヒゲ刺激への応答変化のin vivoイメージング

    橋本 明香里, MIYAMOTO AKIKO, TACHIBANA YOSHIHISA, 春若航一路, WAKE HIROAKI

    関西5医科大学コンソーシアム合宿  2018.9 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:大阪  

  65. 単眼遮蔽による、高次視覚野のヒゲ刺激への応答変化のin vivo イメージング

    橋本 明香里, MIYAMOTO AKIKO, TACHIBANA YOSHIHISA, 春若航一路, WAKE HIROAKI

    第41回日本神経科学大会  2018.7 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:神戸  

  66. 全身炎症時におけるミクログリアによる血液脳関門の制御

    春若航一路, WAKE HIROAKI

    シグナル伝達医学研究展開センター若手道場  2019.1 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:淡路  

  67. 全身炎症時におけるミクログリアによる血液脳関門の制御

    春若航一路, WAKE HIROAKI

    第6回先進イメージング医学研究会・学術集会  2019.1 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:神戸  

  68. Microglia sense systemic immune status to modify activity of neuronal circuit

    Koichiro Haruwaka, Junichi Nabekura, Wake Hiroaki

    第95回日本生理学会大会  2018.3 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:香川  

  69. 光による高次脳機能の計測と操作を目指して

    WAKE HIROAKI

    レーザー学会第39回年次大会  2019.1 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:東京  

  70. 光で迫る脳免疫細胞の機能について Invited

    Wake Hiroaki

    第49回藤田学園医学会シンポジウム(愛知県豊名市)  2017.10 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:愛知  

  71. 光で迫る脳免疫細胞の機能 Invited

    Wake Hiroaki

    第40回日本神経科学大会  2017.7 

     More details

    Language:Japanese   Presentation type:Symposium, workshop panel (nominated)  

    Venue:千葉  

  72. 術後痛モデルマウスにおける第一次体性感覚野 in vivo カルシウムイメージング

    岡田 卓也, 橘 吉寿, NOMURA YUKI, OBATA NORIHIKO, MIZOBUCHI SATOSHI, WAKE HIROAKI

    第41回日本神経科学大会  2018.7 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:神戸  

  73. 身体疾患で惹起される免疫変容が起こす神経回路恒常性の破綻と精神症状の解明

    Wake Hiroaki

    さきがけ領域会議  2017.6 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:秋田  

  74. 術後痛モデルマウスにおける第一次体性感覚野のin vivoカルシウムイメージング

    岡田 卓也, TACHIBANA YOSHIHISA, NOMURA YUKI, OBATA NORIHIKO, MIZOBUCHI SATOSHI, WAKE HIROAKI

    第41回日本神経科学大会  2018.7 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:神戸  

  75. 術後痛モデルマウスにおける第一次体性感覚野 in vivo カルシウムイメージング

    岡田 卓也, 橘 吉寿, NOMURA YUKI, OBATA NORIHIKO, MIZOBUCHI SATOSHI, WAKE HIROAKI

    第5回イメージング数理研究会  2018.7 

     More details

    Language:Japanese   Presentation type:Poster presentation  

    Venue:神戸  

  76. 運動学習と関連した白質における脂質の変化が一次運動野の神経活動に及ぼす影響

    西田 一貴, TACHIBANA YOSHIHISA, 佐藤 駿平, 山崎 文義, 瀬藤 光利, WAKE HIROAKI

    第111回 近畿生理学談話会  2018.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:和歌山  

  77. 高精度時空間分解能を持つ光刺激法による脳機能操作

    Wake Hiroaki

    第14回生体イメージング研究会  2017.8 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:京都  

  78. 高精度時空間分解能を持つ光による脳機能計測・操作を目指して

    WAKE HIROAKI

    イメージング数理研究会  2018.7 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:神戸  

  79. 高次脳機能に関わる中枢神経系免疫細胞の生理機能 Invited

    Wake Hiroaki

    東京大学大学院医学研究科  2017.11 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:東京  

  80. 髄鞘制御不全による情報処理異常の可視化 Invited

    Wake Hiroaki

    神経組織培養研究会  2017.10 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:愛知  

  81. 髄鞘制御不全による情報処理異常の可視化

    Wake Hiroaki

    大阪大学生命機能セミナー  2017.7 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:大阪  

  82. 髄鞘のスクラップアンドビルドによる脳情報処理の効率化

    Wake Hiroaki

    次世代脳冬のシンポジウム  2017.12 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:東京  

  83. 髄鞘制御不全による情報処理異常の可視化 Invited

    Wake Hiroaki

    第29回臨床MR脳機能研究会  2017.4 

     More details

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:東京  

  84. 革新的予防・診断・治療法開発に向けたシグナル伝達医学研究

    Wake Hiroaki

    神戸大学先端融合研究環新規プロジェクトキックオフシンポジウム  2017.5 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:神戸  

  85. 生体適合性マイクロチップレーザーを用いた高次脳機能の4次元操作

    Wake Hiroaki

    融合発展促進研究プロジェクト中間実績報告会  2017.4 

     More details

    Language:Japanese   Presentation type:Oral presentation (general)  

    Venue:東京  

  86. 横浜市立大学「神経医科学序説」における講義

    Wake Hiroaki

    神経医科学序説  2017.5 

     More details

    Language:Japanese   Presentation type:Public lecture, seminar, tutorial, course, or other speech  

    Venue:横浜  

  87. Microglia Sense Systemic Immune Status to Modify Activity of Neuronal Circuit

    Koichiro Haruwaka, Wake Hiroaki, Junichi Nabekura

    Glial assembly, The 4th Summer Workshop  2017.6 

     More details

    Language:English   Presentation type:Poster presentation  

    Venue:新潟  

▼display all

KAKENHI (Grants-in-Aid for Scientific Research) 20

  1. ミクログリアによる感覚モダリティーの制御と精神病態への寄与

    Grant number:21H02662  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業  基盤研究(B)

    和氣 弘明

      More details

    Authorship:Principal investigator 

    Grant amount:\17550000 ( Direct Cost: \13500000 、 Indirect Cost:\4050000 )

    本研究では感覚領域間の機能的結合に着目し、感覚喪失時に引き起こされる異種感覚の可塑性のメカニズムをミクログリアのシナプス修飾の観点から明らかにする。視覚遮断マウスモデルを用いて、第一次感覚野→高次視覚野の機能的結合に着目し、その機能的結合および高次視覚野内で起こる神経回路再編のメカニズムをミクログリアの観点から分子基盤を明らかにする。
    本研究では感覚領域間の機能的結合に着目し、感覚喪失時に引き起こされる異種感覚の可塑性のメカニズムをミクログリアのシナプス修飾の観点から明らかにする。視覚遮断マウスモデルを用いて、第一次感覚野(S1)→高次視覚野(V2L)の異種感覚をつなぐ機能的結合に着目し、その機能的結合および高次視覚野内で起こる神経回路再編のメカニズムをミクログリアの観点から分子基盤を明らかにすることを目的とした。これまでS1-V2Lの回路結合に基づく、先天的視覚遮断においてミクログリアがV2Lの抑制性シナプス伝達を阻害、剥がすことによって、興奮性神経細胞のヒゲ刺激に対する応答を増強させることがわかった。さらにこれによって先天系視覚遮断では感覚識別学習能力が向上する一方、成熟期における視覚遮断ではミクログリアによってヒゲ刺激に対するV2L神経細胞の活動が増強するのにも関わらず、感覚識別学習能力が向上せず異なる神経回路メカニズムが存在することを示した。またこのような臨界期を経る神経回路再編のメカニズムについて経時的なイメージングを行うことでその変化を抽出した (Hashimoto et al., in press)。現在、引き続き統合失調症などをはじめとする精神疾患においてこの異種感覚をつなぐ機能的結合強度に着目し、研究を進めている。
    異種感覚の可塑性のメカニズムについてミクログリアの観点から検証し、論文発表に持っていくことができた
    まず異種感覚をつなぐ領域を感覚統合におけるハブと考え、この領域におけるミクログリアを単離しRNA-seqを用いてミクログリアに発現する分子群を感覚の第一次感覚領野のミクログリアと比較する(特に申請者らが着目しているシナプスに関連する因子MMP9やC4など)。またCx3Cr-GFPマウスの神経細胞にtdtomatoを発現させ、ミクログリアシナプス相関を可視化し、数理学的に解析し、その感覚種別依存性があるのかを検証する。さらに感覚識別力を検証するために感覚学習行動を用いて個体計測する。さらに統合失調症モデルとし臨床患者を実際に反映する22q11.2欠失モデルマウス(入手済み)を用いて、このマウスを先天的な視覚遮断になるようにし、この際のS1の活動をファイバーフォトメトリーで検出しながらV2L内の神経細胞集団活動を可視化し、これを相関付ける。またミクログリアを単離し、その遺伝子発現を解析し、シナプスに対する作用を可視化し、これと相関づける。さらに静脈注射によって大規模に脳内の神経細胞にGCaMPを発現させる系(ウィルス作成済み)を用いて脳のマクロイメージングを実施し、これによって領域間結合(とくに感覚領域間結合)に着目してその機能的結合を評価する

  2. 脳と免疫系相互作用の観点からの多階層的研究による精神疾患病態解明

    Grant number:21H04815  2021.4 - 2024.3

    日本学術振興会  科学研究費助成事業  基盤研究(A)

    尾崎 紀夫, 財津 桂, 大野 欽司, 野田 幸裕, 和氣 弘明

      More details

    Authorship:Coinvestigator(s) 

    脳と免疫系相互作用の観点から精神疾患病態解明の達成を目指し、ゲノム変異を同定した患者由来試料と同一変異に基づくモデル細胞・マウスを用いた多階層的研究を実施する。具体的には、患者ゲノム解析及びゲノム変異患者由来末梢血・腸内細菌・死後脳の解析及びiPS細胞由来神経細胞・オルガノイドの解析。新生仔期免疫活性化状態を経た遺伝子改変マウスの行動解析。ミクログリアと神経細胞の相互作用による神経回路病態解明を目指した、モデルマウスのin vivoイメージング解析。腸内細菌と脳の連関解明を目指した、ヒト・マウスの解析。脳病態と末梢病態との関係の明確化を目指した、ヒト・マウスのインフラマソーム解析。
    代表尾崎は、ゲノム因子と免疫系の相互作用による脳病態を解明するため、22q11.2欠失を代表的なモデルとし、①当該バリアント等の精神疾患患者を対象とした全ゲノム解析・臨床表現型解析、②当該バリアント患者由来iPS細胞由来ミクログリアの機能解析、③当該バリアントモデルマウスを対象とした全脳c-fos解析、④当該バリアント患者死後脳収集を引き続き実施した。
    分担和氣は、統合失調症の周産期免疫活性化モデル(胎児期TLR7注入モデル)を確立し、ストレスを付加することによる行動異常の観察に成功、現在神経回路解析を進めている。一方で感覚統合のメカニズム解明のため、ミクログリアの異種感覚の統合に対する寄与を明らかにし、論文化した。
    分担大野は、22q11.2欠失を有する患者、睡眠障害や摂食障害の患者便からDNA抽出後、MiSeqを用いて16S rRNA V3-V4のamplicon-seqを行い、QIIME2パイプラインによる細菌叢解析を実施。さらに便中短鎖脂肪酸分析により摂食障害の患者便ではイソ酪酸およびイソ吉草酸が上昇しており、腸内細菌叢の変化が示唆された。
    分担野田は、ポリイノシン:ポリシチジル酸(PolyI:C)を投与した仔マウスにおいて増加していたプロスタグランジンE2(PGE2)の高次脳機能障害への影響を検討した。その結果、PolyI:Cによる新生仔期免疫活性化をおこなったマウスにおいて、TNF-αやIL-6、MCP-1、COX-2の遺伝子発現が増加。PGE2による新生仔期の免疫を活性化させたマウスでは、成体期において社会性、認知機能および感覚情報処理などの高次脳機能の障害が認められた。
    分担財津は、PiTMaPメソッドの見直しを行い、キヌレニン経路代謝物の分析条件を検討した。LPSで炎症惹起したマウスの脳試料等を採取し、腹腔内液中IL-1βの上昇が確認された。
    ゲノム解析・iPS細胞解析・モデルマウス解析・死後脳解析のいずれのパートも新たな検体及びデータが得られ、論文化もできていることから概ね順調に進んでいると考えている。また異種感覚統合についても成果を論文化することができ、さらに研究を進めている。22q11.2欠失モデルマウスの解析については、繁殖に遅れが生じていたため、野生型マウスの脳内免疫活性化したモデルマウス(新生仔期免疫活性化マウス)の分子病態の検討に切り替えた。現在、22q11.2欠失モデルマウスの繁殖が安定し、当該マウスに対する新生仔期免疫活性化を順次実施し行動解析を進めている。次年度中旬までに全ての解析を終えることを目標に計画することでこれまでの遅れを取り戻していく予定である。
    代表尾崎は、精神疾患患者のゲノム・血液・死後脳・臨床表現型については引き続き収集し、適宜解析を進める。22q11.2欠失等の発症に関連するゲノムバリアントを有する患者からはiPS細胞を樹立するとともに、表現型解析についても進めていく。モデルマウスについても空間トランスクリプトーム解析等新しい研究手法を取り入れながら機能解析を進めていく。
    分担和氣は、今後異種感覚の統合領域における免疫学的なミクログリアのシナプスに対する寄与を明らかにする。さらに免疫学的な観点から統合失調症の免疫モデル(胎児期TLR7注入モデル)の感覚統合に対するメカニズムの変化を明らかにしていく。
    分担大野は、22q11.2欠失を有する患者、睡眠障害、摂食障害の患者において変化をする腸内細菌叢と短鎖脂肪酸の変化の臨床的な意義づけを今後探る。
    分担野田は、22q11.2欠失モデルマウスの新生仔期に脳内免疫系を活性化させ、各種行動試験を用いて情動・社会性・認知機能・感覚情報処理機能を計画にしたがって検討する予定である。

  3. Effects of maternal undernutrition on offspring brain development via a newly discovered GABAergic CRH release pathway

    Grant number:21H02661  2021.4 - 2024.3

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

      More details

    Authorship:Coinvestigator(s) 

  4. Glia decoding: deciphering information critical for brain-body interactions

    Grant number:20H05894  2020.11 - 2025.3

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

      More details

    Authorship:Coinvestigator(s) 

  5. 全身臓器の生理的・病理的免疫状態遷移の脳による検出機構

    Grant number:20H05899  2020.11 - 2025.3

    日本学術振興会  科学研究費助成事業  学術変革領域研究(A)

    和氣 弘明, 足澤 悦子

      More details

    Authorship:Principal investigator 

    Grant amount:\103220000 ( Direct Cost: \79400000 、 Indirect Cost:\23820000 )

    「全身臓器の情報がグリアインターフェイスによってどのように中枢神経系に伝えられるか」の科学的問いに答えるために脳環境と体循環系環境の境界領域に着目し、境界領域における体循環系免疫細胞と脳免疫細胞の相互作用を生体イメージングを主とした方法で解き明かし、これによって臓器情報がどのように中枢神経系に伝えられるかを明らかにする。
    SLEモデルであるTLR7誘導性SLEモデル(TLR7モデル)を使用し、SLEがウィルス感染などに伴って増悪することに着目し、TLR7モデルにPoly I:C投与を行い、行動変化と脳内炎症の有無を検討した(TLR7+Poly I:Cモデル)。TLR7+Poly I:Cモデルにおいても脾腫がみられ、SLE様症状を呈することが確認された。TLR7+Poly I:Cモデルはコントロール群(Vehicle群)と比較し、オープンフィールドテストで有意に中央滞在率が低下し、不安様症状を呈した。さらに、大脳皮質において、Poly I:C単独投与群(Vehicle+Poly I:C群)と比較して有意なCD3+CD11b-CD45+細胞数の増加がみられた。次に、CD3+細胞の動員にミクログリアが関与しているか検討するため、ミクログリア除去時のCD3+細胞数を解析した。Poly I:C投与の1週間前よりPLX3397(PLX)の投与を行い、ミクログリアを除去した。ミクログリア除去においてVehicle群と比較して有意に活動性の低下がみられ、TLR7+Poly I:Cモデルよりも低下の程度が大きかった。また、大脳皮質のCD3+CD11b-CD45+細胞数は、TLR7+Poly I:Cモデルと比較し、ミクログリア除去で有意に増加していた。CD4/CD8比は有意に低下し、TLR7+Poly I:Cモデルと比較してミクログリア除去でCD4+細胞数は有意差がないが、CD8+細胞数は有意に増加した。以上の結果から、TLR7+Poly I:Cモデルにおいて、大脳皮質でミクログリアがCD8+細胞の浸潤を抑制している可能性が示唆された。CD8+細胞の浸潤はループス腎炎における組織損傷に影響することが知られ、ミクログリアはSLEにおける脳実質の組織損傷から保護していることが考えられた。
    新規SLEモデルの解析が順調に進んでる
    今後は、生体内イメージング技術を用いて、ミクログリアの局性、血液脳関門の透過性の変化を経時的に観察し、解析する。また、蛍光標識したリンパ球を移入し、脳実質内へ浸潤する過程を可視化する。ミクログリアがCD8+細胞を他細胞種と区別し検出する相互作用の挙動を明らかにする。また、RNA-seqによって、TLR7+Poly I:Cモデルとコントロール、TLR7モデルにおけるミクログリアの遺伝子発現解析を行う。さらに先行研究 (Haruwaka et al., 2019)で、ミクログリアは全身炎症に伴って脳血管周囲に遊走し、血液脳関門の透過性に寄与することを明らかにした。この研究の過程で、炎症が消退したのち、ミクログリアは血管周囲から離れ、再配置されることがわかった。この際に炎症時の血液脳関門の透過性増大によって血管内に存在していた蛍光色素を取り込んで、再配置されることがわかった。これはミクログリアを生体で標識することが可能であることを意味する。そこで、蛍光色素として、磁性コーティングされた量子ドットを用いて、ミクログリアを再配置させ、これをMRIで撮像させることを目指した。リポ多糖類で全身炎症を起こしたマウスの静脈内に磁性コーティングされた量子ドットを注入し、ミクログリアに特異的に取り込ませることに成功した。さらに炎症が治った後の再配置されたミクログリア内に量子ドットが取り込まれているのを生体2光子顕微鏡で可視化し、明らかにしたのちに、マウスMRIを撮像した。この状態で1細胞レベルでの検出は困難であることがわかったため、現在7Tの小動物用MRIを用いて、多発性硬化症モデル、およびアルツハイマー型認知症モデルでの検出検証を行っている。

  6. 感覚モダリティ理解のためのミクログリア・シナプス接触の多角的解析

    Grant number:20KK0170  2020.10 - 2024.3

    日本学術振興会  科学研究費助成事業  国際共同研究加速基金(国際共同研究強化(B))

    和氣 弘明, 大野 伸彦, 加藤 大輔, 竹田 育子

      More details

    Authorship:Principal investigator 

    Grant amount:\18720000 ( Direct Cost: \14400000 、 Indirect Cost:\4320000 )

    本国際研究では中枢神経系唯一の免疫細胞であるミクログリアに着目し、ミクログリアがシナプスの構造的・機能的可塑性を修飾する背景を踏まえ、多角的階層的な技術を相互補完することによって、ミクログリアのシナプスに対する時間的(発達・成熟)および空間(脳領域)特異的な生理機能を明らかにする。さらにこれを異種感覚の可塑的変化のメカニズムに繋げ、そこから精神病態を考察する。
    本研究では感覚領域間の機能的結合に着目し、感覚喪失時に引き起こされる異種感覚の可塑性のメカニズムをミクログリアのシナプス修飾の観点から明らかにする。視覚遮断マウスモデルを用いて、第一次感覚野(S1)→高次視覚野(V2L)の異種感覚をつなぐ機能的結合に着目し、その機能的結合および高次視覚野内で起こる神経回路再編のメカニズムをミクログリアの観点から分子基盤を明らかにすることを目的とした。これまでS1-V2Lの回路結合に基づく、先天的視覚遮断においてミクログリアがV2Lの抑制性シナプス伝達を阻害、剥がすことによって、興奮性神経細胞のヒゲ刺激に対する応答を増強させることがわかった。さらにこれによって先天系視覚遮断では感覚識別学習能力が向上する一方、成熟期における視覚遮断ではミクログリアによってヒゲ刺激に対するV2L神経細胞の活動が増強するのにも関わらず、感覚識別学習能力が向上せず異なる神経回路メカニズムが存在することを示した。さらに共同研究者のAnne Schaefer博士とミクログリアの分子情報を比較することで、その分子基盤の一部を明らかにすることに成功した。またこのような臨界期を経る神経回路再編のメカニズムについて経時的なイメージングを行うことでその変化を抽出した (Hashimoto et al., in press)。現在、引き続き統合失調症などをはじめとする精神疾患においてこの異種感覚をつなぐ機能的結合強度に着目し、研究を進め、得られた分子情報を統合失調症モデルのミクログリアの分子情報と比較することでその病態基盤の抽出を行う。
    国際共同研究を論文化することに成功し、順調に進捗している
    まず異種感覚をつなぐ領域を感覚統合におけるハブと考え、この領域におけるミクログリアを単離しRNA-seqを用いてミクログリアに発現する分子群を感覚の第一次感覚領野のミクログリアと比較する(特に研究代表者らが着目しているシナプスに関連する因子MMP9やC4など)。またCx3Cr-GFPマウスの神経細胞にtdtomatoを発現させ、ミクログリアシナプス相関を可視化し、数理学的に解析し、その感覚種別依存性があるのかを検証する。さらに感覚識別力を検証するために感覚学習行動を用いて個体計測する。さらに統合失調症モデルとし臨床患者を実際に反映する22q11.2欠失モデルマウス(入手済み)を用いて、このマウスを先天的な視覚遮断になるようにし、この際のS1の活動をファイバーフォトメトリーで検出しながらV2L内の神経細胞集団活動を可視化し、これを相関付ける。またミクログリアを単離し、その遺伝子発現を解析し、シナプスに対する作用を可視化し、これと相関づける。さらに静脈注射によって大規模に脳内の神経細胞にGCaMPを発現させる系(ウィルス作成済み)を用いて脳のマクロイメージングを実施し、これによって領域間結合(とくに感覚領域間結合)に着目してその機能的結合を評価する。

  7. Development of cell function editing technology with fast 3D measurement and local-field light perturbation

    Grant number:20K20379  2020.4 - 2023.3

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

    Mimori-Kiyosue Yuko

      More details

    Authorship:Coinvestigator(s) 

    The objective of this research project was to construct a "lattice light-sheet microscope" and to introduce cell manipulation techniques by optical stimulation to micro area. Using the constructed microscope, we succeeded in visualizing the three-dimensional dynamics of the cytoskeleton in cell division and migration, and the process by which extracellular vesicles are generated by cleavage of cell protrusions. The optical stimuli were irradiated to targeted submicron to several micron regions by sweeping Bessel beams. In addition, using two-photon microscopy, excitation light was created in living organisms in ~100 multi-spots by holographic optical pathways, and the fluorescence generated was successfully detected at 100-400 Hz. We are working to apply these technologies to cells while advancing their sophistication.

  8. Local response of oligodendrocyte progenitor cell regulate axonal activity

    Grant number:19H04753  2019.4 - 2021.3

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research  Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

      More details

    Authorship:Principal investigator 

    Grant amount:\9490000 ( Direct Cost: \7300000 、 Indirect Cost:\2190000 )

  9. 免疫異常の記憶から捉える精神疾患の階層的理解

    Grant number:19H05219  2019.4 - 2021.3

    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)

    和氣 弘明

      More details

    Authorship:Principal investigator 

    Grant amount:\7540000 ( Direct Cost: \5800000 、 Indirect Cost:\1740000 )

    大脳皮質感覚野に着目し、各発達期において免疫異常の履歴をもつマウスのエピジェネティックな変化および遺伝子発現の変化を、DNAメチル化に対する検証およびミクログリア特異的遺伝子発現解析法を用いて明らかにし、成熟過程のミクログリアを2光子顕微鏡による機能イメージングと相関づける。続いて、この免疫履歴をもつマウスに対して拘束ストレスを付加し、誘導されるミクログリアの機能変化を検出し、その後表出する遺伝子異常と結びつける。さらにミクログリアの機能変化によって引き起こされる表現型の差異を、特にミクログリアによるシナプス修飾・血液脳関門の透過性制御に着目して、2光子顕微鏡で可視化することで明らかにする。
    本研究ではミクログリアによる精神疾患の関与を解明する研究を行った。ミクログリアはこれまで成熟期においてシナプスなどの恒常性を維持する働きがあることがわかっている。近年全身炎症などが精神症状に寄与することが知られているが、この全身炎症に伴ってミクログリアが血管にCCL5-CCR5のシグナルによって遊走し、炎症早期はCldn5を発現することによって血液脳関門(BBB)の透過性に保護的に作用すること炎症後期においてはCD68を発現することによって、BBBの透過性を増加させることを明らかにした。これによってBBBを介した精神疾患の治療戦略を組み立てることができるようになった。さらに妊娠中のマウスに炎症を引き起こすことで児の自閉症を引き起こすことが知られているが、この際に増加するサイトカインプロファイルを明らかにし、ミクログリアの成熟を促すマーカーの変化を伴うこと、さらにこの極性が行動異常と相関することから、ミクログリアの動態の極性を誘導することで、その生理機能を損なうことを明らかにした。また2Hit論理で統合失調症様症状を発症するモデルマウスの創出に成功し、このマウスにおけるミクログリアのエピゲノム変化が成熟期の発現分子変化およびシナプスに対する作用を変化させることを長期生体イメージングを用いて明らかにし、さらにこのエピゲノムの変化がストレスによって行動異常をもたらす分子基盤及び神経回路基盤を明らかにした。
    令和2年度が最終年度であるため、記入しない。
    令和2年度が最終年度であるため、記入しない。

  10. Microglial contribution on higher order of brain function and their pathology

    Grant number:18H02598  2018.4 - 2021.3

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

    WAKE Hiroaki

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\17420000 ( Direct Cost: \13400000 、 Indirect Cost:\4020000 )

    In this study, we focused on microglia and aimed to elucidate psychiatric pathology from their physiological functions on synapses and blood vessels. Using in vivo imaging, we found that microglia migrate into cerebral blood vessels during systemic inflammation and contribute to the permeability of the blood-brain barrier in a time-specific manner by altering the expression of tight junction-related factors and phagocytic factors. In addition, we found that microglia modulate their synaptic effects by regulating their projection dynamics during higher brain functions (motor learning). These synaptic modulation mechanisms are dependent on the sensory input in the sensory cortex, suggesting a role for microglia in heterogeneous sensory plasticity.

  11. Therapeutic strategy for the abnormal neuronal circuit activity associated with systemic inflammation

    Grant number:17K19458  2017.6 - 2019.3

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

    WAKE Hiroaki

      More details

    Authorship:Principal investigator 

    Grant amount:\6500000 ( Direct Cost: \5000000 、 Indirect Cost:\1500000 )

    Microglia survey brain parenchyma and respond to any disruptions. Microglia also respond to systemic disease, but how this relates to blood brain barrier (BBB) integrity is largely unknown. Here we use simultaneous in vivo imaging to demonstrate that systemic inflammation induces migration of brain resident microglia to cerebral vessels. Vessel-associated microglia initially maintain BBB integrity, associated with expression of the tight junction protein Claudin-5. Further sustained inflammation results in microglia phagocytosing astrocytic end-feet and impairing BBB function. Our results show dual microglial role for BBB and have important implications for understanding how systemic immune-activation can impact on neural circuit functions.

  12. オリゴデンドロサイトの制御による神経科回路活動の精緻化

    Grant number:17H05747  2017.4 - 2019.3

    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)

    和氣 弘明

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\11050000 ( Direct Cost: \8500000 、 Indirect Cost:\2550000 )

    オリゴデンドロサイトは髄鞘を形成する細胞で、これまで神経活動依存的に髄鞘化することが知られている。これまで私たちはオリゴデンドロサイトが神経活動依存的に髄鞘化する分子メカニズムを示し、さらに軸索との結合様式を明らかにしてきた。本研究ではこの神経活動依存的髄鞘化を担う、オリゴデンドロサイトの活動を生体で可視化するとともに、その髄鞘を構成する脂質成分を捉えるべく研究を行った。
    1.生体でオリゴデンドロサイトおよびその前駆細胞を可視化するために、PLPプロモーターの下流にカルシウム感受性蛍光タンパク質がテトラサイクリン依存的に発現するマウスを用いて、これを2光子顕微鏡によって生体覚醒下で可視化した。神経細胞活動を操作するために覚醒下、麻酔下の2条件で比較した。細胞体のカルシウム上昇頻度はオリゴデンドロサイトおよびその前駆細胞すべての例において覚醒下氏比して麻酔下では有意に低下した。さらにその突起の活動も有意に低下することを明らかにした。一方でDREAAD法を用いて神経細胞活動を上昇させたところ細胞体の活動は大きく変化せず、突起活動は突起によるバリエーションを認めた。この中からカルシウム上昇が高頻度に起こる部位をホットスポットと定義し、ホットスポットにおけるカルシウム上昇頻度が有意に上昇するのを認めた。またこのホットスポットが学習などの負荷によって高頻度にターンオーバーすることがわかり、現在結果をまとめている。またこのような髄鞘は脂質で構成されていることが知られている。このような脂質変化を神経回路活動の変化と結びつけるため、マウスにレバー引き水報酬学習を行わせ、神経細胞集団の活動を可視化した後、質量分析顕微鏡で脂質成分の変化を検出し、その相関を得られることができた。
    平成30年度が最終年度であるため、記入しない。
    平成30年度が最終年度であるため、記入しない。

  13. 全身免疫異常に伴う神経回路活動の変容に対する治療戦略

    2017.4 - 2019.3

    学術研究助成基金助成金/挑戦的研究(萌芽) 

    和氣 弘明

      More details

    Authorship:Principal investigator  Grant type:Competitive

  14. ホログラム光刺激による神経回路再編の人為的創出

    2017

    国立研究開発法人科学技術振興機構  戦略的創造研究推進事業(CREST) 

    和氣 弘明

      More details

    Authorship:Principal investigator  Grant type:Competitive

  15. ミクログリアによるシナプス活動修飾と神経回路の空間的活動制御

    Grant number:16H01346  2016.4 - 2018.3

    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)

    和氣 弘明

      More details

    Authorship:Principal investigator 

    Grant amount:\9490000 ( Direct Cost: \7300000 、 Indirect Cost:\2190000 )

    本課題では、感覚刺激、運動学習において生理学的なミクログリア突起の動きによるシナプスへの接触でシナプス活動ひいては神経回路活動にどのように影響し、シナプス機能、可塑性、数を制御するかを検証する。次にこの制御機構の破綻で発達障害、精神・神経疾患を惹起しうるかを考える。これまで、2光子顕微鏡を用いた生体イメージングによってシナプス活動とミクログリアを同時に可視化し、ミクログリアの接触時にシナプス活動が増加することを示し、さらにこのミクログリアによるシナプス活動の修飾はミクログリアをリポ多糖類で活性化させることで消失することを明らかにした。またミクログリアをジフテリア毒素によって遺伝的に時期特異的に除去できるマウス(Iba1-tTa::TetO-DTA)を用いることで、ミクログリアを除去すると神経細胞活動の同期性が減少することから、ミクログリアによるシナプス活動の修飾は神経細胞集団の同期性に寄与していることを示した(論文投稿中)。またミクログリアがシナプス活動を修飾するのであれば、学習過程によってその動態が変化し、シナプス活動の修飾機構が変化する可能性がある。そこでレバー引きによる水報酬学習を用いてその前後でミクログリア突起の動態の比較をしたところ、学習前に比べて、学習後ではミクログリア突起の動態の複雑性が低下し、より定型的になっていることを明らかにした。またこのようなミクログリア突起動態の変化が精神疾患などのモデルでも認められるかどうか検証するために統合失調症モデル(Shn2-KO)で同様に比較したところ統合失調症モデルではミクログリア突起の動態の複雑性が増していることがわかった。そこで、これらを踏まえ、今後ミクログリアのシナプス修飾機構から精神疾患を考察していく。
    29年度が最終年度であるため、記入しない。
    29年度が最終年度であるため、記入しない。

  16. 身体疾患で惹起される免疫変容が起こす神経回路恒常性の破綻と精神症状の解明

    2016

    国立研究開発法人科学技術振興機構  戦略的創造研究推進事業(さきがけ) 

    和氣 弘明

      More details

    Authorship:Principal investigator  Grant type:Competitive

  17. ミクログリアによるシナプス活動修飾と神経回路の空間的活動制御

    2015.4 - 2018.3

    科学研究費補助金/新学術領域研究 

    和氣 弘明

      More details

    Authorship:Principal investigator  Grant type:Competitive

  18. グリア細胞からみる精神疾患

    Grant number:15H01300  2015.4 - 2017.3

    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)

    和氣 弘明

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\4680000 ( Direct Cost: \3600000 、 Indirect Cost:\1080000 )

    多様化する現代社会において、高次脳機能に異常を呈する発達障害・精神疾患の病態の解明および治療法の開発は喫緊の課題である。本課題では統合失調症モデルマウスにおける神経回路基盤の変化を脳内免疫と結びつけて検証する。すなわち統合失調症モデルとしてSchnurri-2 欠損マウスを用い、Schnurri-2 欠損マウスの中枢神経系炎症反応の増大の結果としてミクログリアにおけるシナプス制御分子、髄鞘制御分子の変化を検出し、これを、2光子顕微鏡によるin vivoイメージングを組み合わせることにより行動異常と神経回路動作の相関を神経細胞の発火パターン、シナプスの活動パターンを可視化することによって明らかにする。これまでSchnurri-2 欠損マウスにおける神経細胞の発火率、発火パターンを検証するためにカルシウム感受性蛍光タンパク質であるGCaMP6fをコードする遺伝子をアデノ随伴ウィルス(AAV)の大脳皮質感覚野・運動野への注入によって発現させ、その神経細胞集団音発火を解析することによって覚醒下の神経細胞活動の協調性の低下が認められていることを明らかにした。さらに中枢神経系免疫細胞であるミクログリアを免疫染色することによってその突起の短縮および複雑性の低下を認めた。さらにSchnurri-2 欠損マウスのミクログリアにGFPが発現するマウスを作成し、2光子顕微鏡を用いて突起の動きを解析したところ、その動きのランダムさが亢進していることがわかった(Haruwaka et al., in preparation)。これはシナプス活動のランダムさを反映するものであると考えている。今後この変化を担う分子の同定を行うとともに、行動との相関を検証する予定である

  19. The effect of activity dependent myelination on to the neuronal circuit activity

    Grant number:26710004  2014.4 - 2018.3

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

    WAKE Hiroaki, KATO Daisuke, AKIYOSHI Ryohei

      More details

    Authorship:Principal investigator  Grant type:Competitive

    Grant amount:\24570000 ( Direct Cost: \18900000 、 Indirect Cost:\5670000 )

    Myelination increases conduction velocity and promotes brain functions. Myelin dysregulation is frequently associated with learning and cognition deficits, ultimately causing neurological and psychiatric disorders. However, it has not been revealed what perturbation of neural activity induced by myelin dysregulation impairs learning. Here, we measured neural activity in the motor cortex during motor learning in transgenic mice with a subtle impairment of their myelin regulation.This myelin dysregulation impaired motor learning and was accompanied by a decrease in the amplitude of movement-related activity, an increase in the frequency of spontaneous activity, and a widening in the timing of cortical responses to thalamic stimulation. Repetitive pairing of forelimb movements with optogenetic stimulation of thalamocortical axon terminals partially restored learning.

  20. スキャンレス3D ホログラフィック計測・刺激顕微鏡の開発と生体応用

    国立研究開発法人科学技術振興機構  A-STEP産学共同【本格型】 

▼display all

 

Teaching Experience (Off-campus) 17

  1. 文献解析・プレゼンテーション演習

    Kobe University)

  2. Medical Science A

    Kobe University)

  3. Literature Analysis and Presentation

    Kobe University)

  4. Forefront of Research at Kobe University B

    Kobe University)

  5. Biomedical ScienceA

    Kobe University)

  6. Basic Course of Bioscience

    Kobe University)

  7. Medical Science B

    Kobe University)

  8. 神戸大学の研究最前線B

    Kobe University)

  9. 生理学

    Kobe University)

  10. 医学B

    Kobe University)

  11. 医学A

    Kobe University)

  12. 医学序説

    Kobe University)

  13. バイオメディカルサイエンス特別研究

    Kobe University)

  14. バイオメディカルサイエンスA

    Kobe University)

  15. バイオサイエンス基本実習

    Kobe University)

  16. Research in Biomedical Science

    Kobe University)

  17. Physiology

    Kobe University)

▼display all