|
職名 |
教授 |
|
外部リンク |
|
|
|
中澤 敬信 (ナカザワ タカノブ) NAKAZAWA Takanobu 教授 |
論文 【 表示 ／ 非表示 】
-
Kazuyuki Togo, Hayato Fukusumi, Tomoko Shofuda, Hiroshi Ohnishi, Hiroyuki Yamazaki, Mariko Kato Hayashi, Nana Kawasaki, Nobuyuki Takei, Takanobu Nakazawa, Yumiko Saito, Kousuke Baba, Hitoshi Hashimoto, Yuko Sekino, Tomoaki Shirao, Hideki Mochizuki, Yonehiro Kanemura
Molecular Brain 14 ( 1 ) 2021年12月
担当区分：筆頭著者 掲載種別：研究論文（学術雑誌） 出版者・発行元：Springer Science and Business Media LLC
<title>Abstract</title>The generation of mature synaptic structures using neurons differentiated from human-induced pluripotent stem cells (hiPSC-neurons) is expected to be applied to physiological studies of synapses in human cells and to pathological studies of diseases that cause abnormal synaptic function. Although it has been reported that synapses themselves change from an immature to a mature state as neurons mature, there are few reports that clearly show when and how human stem cell-derived neurons change to mature synaptic structures. This study was designed to elucidate the synapse formation process of hiPSC-neurons. We propagated hiPSC-derived neural progenitor cells (hiPSC-NPCs) that expressed localized markers of the ventral hindbrain as neurospheres by dual SMAD inhibition and then differentiated them into hiPSC-neurons in vitro. After 49 days of in vitro differentiation, hiPSC-neurons significantly expressed pre- and postsynaptic markers at both the transcript and protein levels. However, the expression of postsynaptic markers was lower than in normal human or normal rat brain tissues, and immunostaining analysis showed that it was relatively modest and was lower than that of presynaptic markers and that its localization in synaptic structures was insufficient. Neurophysiological analysis using a microelectrode array also revealed that no synaptic activity was generated on hiPSC-neurons at 49 days of differentiation. Analysis of subtype markers by immunostaining revealed that most hiPSC-neurons expressed vesicular glutamate transporter 2 (VGLUT2). The presence or absence of NGF, which is required for the survival of cholinergic neurons, had no effect on their cell fractionation. These results suggest that during the synaptogenesis of hiPSC-neurons, the formation of presynaptic structures is not the only requirement for the formation of postsynaptic structures and that the mRNA expression of postsynaptic markers does not correlate with the formation of their mature structures. Technically, we also confirmed a certain level of robustness and reproducibility of our neuronal differentiation method in a multicenter setting, which will be helpful for future research. Synapse formation with mature postsynaptic structures will remain an interesting issue for stem cell-derived neurons, and the present method can be used to obtain early and stable quality neuronal cultures from hiPSC-NPCs.
DOI： 10.1186/s13041-021-00851-1
その他リンク： https://link.springer.com/article/10.1186/s13041-021-00851-1/fulltext.html
-
Intranasal oxytocin administration ameliorates social behavioral deficits in a POGZWT/Q1038R mouse model of autism spectrum disorder.
Kohei Kitagawa, Kensuke Matsumura, Masayuki Baba, Momoka Kondo, Tomoya Takemoto, Kazuki Nagayasu, Yukio Ago, Kaoru Seiriki, Atsuko Hayata-Takano, Atsushi Kasai, Kazuhiro Takuma, Ryota Hashimoto, Hitoshi Hashimoto, Takanobu Nakazawa
Molecular brain 14 ( 1 ) 56 - 56 2021年03月
担当区分：筆頭著者 記述言語：英語 掲載種別：研究論文（学術雑誌）
Autism spectrum disorder (ASD) is a highly prevalent neurodevelopmental disorder characterized by core symptoms of impaired social behavior and communication. Recent studies have suggested that the oxytocin system, which regulates social behavior in mammals, is potentially involved in ASD. Mouse models of ASD provide a useful system for understanding the associations between an impaired oxytocin system and social behavior deficits. However, limited studies have shown the involvement of the oxytocin system in the behavioral phenotypes in mouse models of ASD. We have previously demonstrated that a mouse model that carries the ASD patient-derived de novo mutation in the pogo transposable element derived with zinc finger domain (POGZWT/Q1038R mice), showed ASD-like social behavioral deficits. Here, we have explored whether oxytocin (OXT) administration improves impaired social behavior in POGZWT/Q1038R mice and found that intranasal oxytocin administration effectively restored the impaired social behavior in POGZWT/Q1038R mice. We also found that the expression level of the oxytocin receptor gene (OXTR) was low in POGZWT/Q1038R mice. However, we did not detect significant changes in the number of OXT-expressing neurons between the paraventricular nucleus of POGZWT/Q1038R mice and that of WT mice. A chromatin immunoprecipitation assay revealed that POGZ binds to the promoter region of OXTR and is involved in the transcriptional regulation of OXTR. In summary, our study demonstrate that the pathogenic mutation in the POGZ, a high-confidence ASD gene, impairs the oxytocin system and social behavior in mice, providing insights into the development of oxytocin-based therapeutics for ASD.
-
Methylation Analysis in Monozygotic Twins With Treatment-Resistant Schizophrenia and Discordant Responses to Clozapine.
Masataka Kikuchi, Takanobu Nakazawa, Makoto Kinoshita, Hidenaga Yamamori, Yuka Yasuda, Michiko Fujimoto, Ryota Hashimoto, Shusuke Numata
Frontiers in psychiatry 12 734606 - 734606 2021年
担当区分：筆頭著者 記述言語：英語 掲載種別：研究論文（学術雑誌）
Schizophrenia is a mental illness that involves both genetic and environmental factors. Clozapine, an atypical antipsychotic, is a well-established therapy for treatment-resistant schizophrenia. In this study, we focused on a set of monozygotic twins with treatment-resistant schizophrenia in which one twin effectively responded to clozapine treatment and the other did not. Our previous study generated neurons from induced pluripotent stem (iPS) cells derived from these patients and compared the transcriptome profiles between mock- and clozapine-treated neurons. In this study, we performed genome-wide DNA methylation profiling to investigate the mechanisms underlying gene expression changes. First, we extracted the differentially methylated sites from each twin based on statistical analysis. Then, we combined the DNA methylation profiling with transcriptome profiling from our previous RNA-seq data. Among the genes with altered methylation and expression, we found the different proportions of the genes related to neuronal and synaptic functions between the clozapine responder and non-responder (35.7 and 6.7%, respectively). This trend was observed even when the basal differences between the responder and non-responder was excluded. These results suggest that effective clozapine action may correct the abnormalities of neuronal and synapse functions in schizophrenia via changes in methylation.
-
An Autism-Associated Neuroligin-3 Mutation Affects Developmental Synapse Elimination in the Cerebellum.
Esther Suk King Lai, Hisako Nakayama, Taisuke Miyazaki, Takanobu Nakazawa, Katsuhiko Tabuchi, Kouichi Hashimoto, Masahiko Watanabe, Masanobu Kano
Frontiers in neural circuits 15 676891 - 676891 2021年
担当区分：筆頭著者 記述言語：英語 掲載種別：研究論文（学術雑誌）
Neuroligin is a postsynaptic cell-adhesion molecule that is involved in synapse formation and maturation by interacting with presynaptic neurexin. Mutations in neuroligin genes, including the arginine to cystein substitution at the 451st amino acid residue (R451C) of neuroligin-3 (NLGN3), have been identified in patients with autism spectrum disorder (ASD). Functional magnetic resonance imaging and examination of post-mortem brain in ASD patients implicate alteration of cerebellar morphology and Purkinje cell (PC) loss. In the present study, we examined possible association between the R451C mutation in NLGN3 and synaptic development and function in the mouse cerebellum. In NLGN3-R451C mutant mice, the expression of NLGN3 protein in the cerebellum was reduced to about 10% of the level of wild-type mice. Elimination of redundant climbing fiber (CF) to PC synapses was impaired from postnatal day 10-15 (P10-15) in NLGN3-R451C mutant mice, but majority of PCs became mono-innervated as in wild-type mice after P16. In NLGN3-R451C mutant mice, selective strengthening of a single CF relative to the other CFs in each PC was impaired from P16, which persisted into juvenile stage. Furthermore, the inhibition to excitation (I/E) balance of synaptic inputs to PCs was elevated, and calcium transients in the soma induced by strong and weak CF inputs were reduced in NLGN3-R451C mutant mice. These results suggest that a single point mutation in NLGN3 significantly influences the synapse development and refinement in cerebellar circuitry, which might be related to the pathogenesis of ASD.
-
Pathogenic POGZ mutation causes impaired cortical development and reversible autism-like phenotypes. 査読あり
Kensuke Matsumura, Kaoru Seiriki, Shota Okada, Masashi Nagase, Shinya Ayabe, Ikuko Yamada, Tamio Furuse, Hirotoshi Shibuya, Yuka Yasuda, Hidenaga Yamamori, Michiko Fujimoto, Kazuki Nagayasu, Kana Yamamoto, Kohei Kitagawa, Hiroki Miura, Nanaka Gotoda-Nishimura, Hisato Igarashi, Misuzu Hayashida, Masayuki Baba, Momoka Kondo, Shigeru Hasebe, Kosei Ueshima, Atsushi Kasai, Yukio Ago, Atsuko Hayata-Takano, Norihito Shintani, Tokuichi Iguchi, Makoto Sato, Shun Yamaguchi, Masaru Tamura, Shigeharu Wakana, Atsushi Yoshiki, Ayako M Watabe, Hideyuki Okano, Kazuhiro Takuma, Ryota Hashimoto, Hitoshi Hashimoto, Takanobu Nakazawa
Nature communications 11 ( 1 ) 859 - 859 2020年02月
担当区分：筆頭著者 記述言語：英語 掲載種別：研究論文（学術雑誌）
Pogo transposable element derived with ZNF domain (POGZ) has been identified as one of the most recurrently de novo mutated genes in patients with neurodevelopmental disorders (NDDs), including autism spectrum disorder (ASD), intellectual disability and White-Sutton syndrome; however, the neurobiological basis behind these disorders remains unknown. Here, we show that POGZ regulates neuronal development and that ASD-related de novo mutations impair neuronal development in the developing mouse brain and induced pluripotent cell lines from an ASD patient. We also develop the first mouse model heterozygous for a de novo POGZ mutation identified in a patient with ASD, and we identify ASD-like abnormalities in the mice. Importantly, social deficits can be treated by compensatory inhibition of elevated cell excitability in the mice. Our results provide insight into how de novo mutations on high-confidence ASD genes lead to impaired mature cortical network function, which underlies the cellular pathogenesis of NDDs, including ASD.
科研費（文科省・学振）獲得実績 【 表示 ／ 非表示 】
-
患者iPS細胞由来脳オルガノイド及び疾患モデルマウスを用いた自閉症の分子病態研究
2021年04月 - 2024年03月
科学研究費補助金 基盤研究(B)
中澤敬信
-
iPS細胞技術及び患者臨床情報を用いた精神疾患の分子病態の多階層解析
2021年04月 - 2023年03月
科学研究費補助金 新学術領域研究
中澤敬信
-
社会行動制御に関わる神経細胞の単一細胞レベルの解析及びその活動操作の基盤技術開発
2020年10月 - 2022年03月
科学研究費補助金 挑戦的萌芽研究
中澤 敬信
担当区分：研究代表者
その他競争的資金獲得実績 【 表示 ／ 非表示 】
-
自閉症における社会性相互作用障害に注目した環境要因の中枢分子薬理学研究
2021年04月 - 2025年03月
旭硝子財団
中澤敬信
資金種別：競争的資金
-
分子病態に基づく自閉症の分類化のトランスレーショナル研究
2020年10月 - 2022年10月
武田科学振興財団
中澤 敬信
担当区分：研究代表者 資金種別：競争的資金
-
自閉症に高頻度に認めるPOGZ遺伝子変異に注目した自己と他者との社会的相互作用制御の分子基盤解明
2018年 - 2019年
旭硝子財団
中澤 敬信、中澤敬信
担当区分：研究代表者 資金種別：競争的資金
受託研究受入実績 【 表示 ／ 非表示 】
-
iPS細胞技術とデータ科学を融合した精神疾患横断的な双方向トランスレーショナル研究
2021年07月 - 2025年03月
日本医療研究開発機構（AMED） 一般受託研究 一般受託研究
橋本亮太
-
神経発達障害の病態解明を目指した革新的イメージングプラットフォーム
2019年10月 - 2025年03月
日本医療研究開発機構（AMED） 一般受託研究 一般受託研究
岡部繁男
担当区分：研究代表者
-
治療抵抗性統合失調症に対する客観的診断法及び実用的治療プロトコールの開発
2016年04月 - 2017年03月
日本医療研究開発機構（AMED） 一般受託研究 一般受託研究
橋本亮太
担当区分：研究代表者
担当経験のある科目（授業） 【 表示 ／ 非表示 】
-
分子生物学（一）
機関名：東京農業大学
-
分子遺伝学
機関名：東京農業大学
-
臨床薬理学演習
機関名：大阪大学歯学部
-
臨床薬理学講義
機関名：大阪大学歯学部
-
先端生命科学特別講義
機関名：大阪大学大学院薬学研究科