Cell type specific analysis of intracellular signal in neural circuit

• Project/Area Number: 15K06772
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Neurochemistry/Neuropharmacology
• Research Institution: Nagoya University
• Principal Investigator: Kuroda Keisuke 名古屋大学, 医学系研究科, 特任助教 (80631431)
• Project Period (FY): 2015-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 細胞内情報伝達 / シグナル解析 / 細胞種特異的 / リン酸化 / アデノ随伴ウイルス / ドーパミン / アデノシン / 側坐核 / 神経化学 / Rho / AAV-Flexシステム

Outline of Final Research Achievements

Dopamine activates PKA via D1 receptor / Gαs pathway in D1R-MSN of the nucleus accumbens. On the other hand, the adenosine / A2A receptor / Gαs pathway and the dopamine / D2 receptor / Gαi pathway are antagonized in D2R-MSN. The D2 receptor is constantly activated by dopamine, and when dopamine secretion decreases, repression is released and PKA is activated in D2R-MSN. Activated PKA phosphorylates and activates RasGRP2, which is an activator of Rap1 (GEF), and phosphorylates and inactivates Rap1GAP, which is an inactivator of Rap1 (GAP). PKA activates Rap1 / MAPK signals and enhances neuronal excitability.

Academic Significance and Societal Importance of the Research Achievements

抗精神病薬や抗うつ薬がドーパミンやセロトニンなどのモノアミンの作用を制御していることはよく知られているが、脳内においてどのような分子メカニズムによってその作用を発揮しているかは驚くほどわかっていない。本研究により側坐核の中型有棘神経細胞においてドーパミン受容体の下流で低分子量G蛋白質Rap1が神経細胞の興奮性を制御することで、情動を生み出すスイッチとして働いていることが明らかになった。本研究のようなアプローチで細胞内シグナル伝達機構を解明することによって、新たな治療ターゲットの発見に繋がると考えられる。

Research Products

• [Journal Article] Repetitive and compulsive-like behaviors lead to cognitive dysfunction in Disc1Δ2-3/Δ2-3 mice (2018)
  • Author(s): Wulaer B, Nagai T, Sobue A, Itoh N, Kuroda K, Kaibuchi K, Nabeshima T, Yamada K
  • Journal Title: Genes Brain Behav Volume: 印刷中 Issue: 8
  • DOI: 10.1111/gbb.12478
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cell type-specific activation of mitogen-activated protein kinase in D1 receptor-expressing neurons of the nucleus accumbens potentiates stimulus-reward learning in mice (2018)
  • Author(s): Bin Saifullah Md. Ali、Nagai Taku、Kuroda Keisuke、Wulaer Bolati、Nabeshima Toshitaka、Kaibuchi Kozo、Yamada Kiyofumi
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 14413-14413
  • DOI: 10.1038/s41598-018-32840-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Balance between dopamine and adenosine signals regulates the PKA/Rap1 pathway in striatal medium spiny neurons (2018)
  • Author(s): Zhang XJ, Nagai T, Ahammad RU, Kuroda K, Nakamuta S, Nakano T, Yukinawa N, Funahashi Y, Yamahashi Y, Amano M, Yoshimoto J, Yamada K, and Kaibuchi K.
  • Journal Title: Neurochemistry International Volume: 122 Pages: 8-18
  • DOI: 10.1016/j.neuint.2018.10.008
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Comprehensive analysis of kinase-oriented phospho-signalling pathways (2018)
  • Author(s): Amano Mutsuki、Nishioka Tomoki、Tsuboi Daisuke、Kuroda Keisuke、Funahashi Yasuhiro、Yamahashi Yukie、Kaibuchi Kozo
  • Journal Title: The Journal of Biochemistry Volume: 165 Issue: 4 Pages: 301-307
  • DOI: 10.1093/jb/mvy115
  • Peer Reviewed / Open Access
• [Journal Article] Neuropeptide Y neuronal network dysfunction in the frontal lobe of a genetic mouse model of schizophrenia (2017)
  • Author(s): Morosawa S, Iritani S, Fujishiro H, Sekiguchi H, Torii Y, Habuchi C, Kuroda K, Kaibuchi K, Ozaki N.
  • Journal Title: Neuropeptides Volume: 16 Pages: 30121-30124
  • DOI: 10.1016/j.npep.2016.12.010
  • Peer Reviewed
• [Journal Article] Focused Proteomics Revealed a Novel Rho-kinase Signaling Pathway in the Heart (2016)
  • Author(s): Yura Y, Amano M, Takefuji M, Bando T, Suzuki K, Kato K, Hamaguchi T, Hasanuzzaman Shohag M, Takano T, Funahashi Y, Nakamuta S, Kuroda K, Nishioka T, Murohara T, Kaibuchi K
  • Journal Title: Cell Structure and Function Volume: 41 Issue: 2 Pages: 105-120
  • DOI: 10.1247/csf.16011
  • NAID: 130005405432
  • ISSN: 0386-7196, 1347-3700
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Stimulation of Synaptic Vesicle Exocytosis by the Mental Disease Gene DISC1 is Mediated by N-Type Voltage-Gated Calcium Channels (2016)
  • Author(s): Tang W, Thevathasan JV, Lin Q, Lim KB, Kuroda K, Kaibuchi K, Bilger M, Soong TW, Fivaz M
  • Journal Title: Frontiers in Synaptic Neuroscience Volume: 8
  • DOI: 10.3389/fnsyn.2016.00015
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Survival of corticostriatal neurons by Rho/Rho-kinase signaling pathway. (2016)
  • Author(s): Kobayashi K, Sano H, Kato S, Kuroda K, Nakamuta S, Isa T, Nambu A, Kaibuchi K, Kobayashi K.
  • Journal Title: Neurosci Lett. Volume: 630 Pages: 45-52
  • DOI: 10.1016/j.neulet.2016.07.020
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Phosphorylation Signals in Striatal Medium Spiny Neurons (2016)
  • Author(s): Nagai T, Yoshimoto J, Kannon T, Kuroda K, Kaibuchi K
  • Journal Title: Trends in Pharmacological Sciences Volume: 37 Issue: 10 Pages: 858-71
  • DOI: 10.1016/j.tips.2016.07.003
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Immunohistochemical evaluation of the GABAergic neuronal system in the prefrontal cortex of a DISC1 knockout mouse model of schizophrenia (2016)
  • Author(s): Umeda K, Iritani S, Fujishiro H, Sekiguchi H, Torii Y, Habuchi C, Kuroda K, Kaibuchi K, Ozaki N.
  • Journal Title: Synapse Volume: 70 Issue: 12 Pages: 508-518
  • DOI: 10.1002/syn.21924
  • Peer Reviewed
• [Journal Article] Catecholaminergic neuronal network dysfunction in the frontal lobe of a genetic mouse model of schizophrenia (2016)
  • Author(s): Shuji Iritani, Hirotaka Sekiguchi, Chikako Habuchi, Youta Torii, Keisuke Kuroda, Kozo Kaibuchi and Norio Ozaki
  • Journal Title: Acta Neuropsychiatrica Volume: 28 Issue: 2 Pages: 117-123
  • DOI: 10.1017/neu.2015.51
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Phosphoproteomics of the Dopamine Pathway Enables Discovery of Rap1 Activation as a Reward Signal In Vivo (2016)
  • Author(s): Nagai T, Nakamuta S, Kuroda K, Nakauchi S, Nishioka T, Takano T, Zhang X, Tsuboi D, Funahashi Y, Nakano T, Yoshimoto J, Kobayashi K, Uchigashima M, Watanabe M, Miura M, Nishi A, Kobayashi K, Yamada K, Amano M, Kaibuchi K
  • Journal Title: Neuron Volume: 89 (3) Issue: 3 Pages: 550-65
  • DOI: 10.1016/j.neuron.2015.12.019
  • Peer Reviewed / Open Access
• [Journal Article] Disrupted-in-schizophrenia-1 (DISC1) Regulates Endoplasmic Reticulum Calcium Dynamics. (2015)
  • Author(s): Park SJ, Jeong J, Park YU, Park KS, Lee H, Lee N, Kim SM, Kuroda K, Nguyen MD, Kaibuchi K, Park SK.
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 8694-8694
  • DOI: 10.1038/srep08694
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Disrupted-in-schizophrenia 1 regulates transport of ITPR1 mRNA for synaptic plasticity. (2015)
  • Author(s): Tsuboi D, Kuroda K, Tanaka M, Namba T, Iizuka Y, Taya S, Shinoda T, Hikita T, Muraoka S, Iizuka M, Nimura A, Mizoguchi A, Shiina N, Sokabe M, Okano H, Mikoshiba K, Kaibuchi K
  • Journal Title: Nature Neuroscience Volume: in press Issue: 5 Pages: 698-707
  • DOI: 10.1038/nn.3984
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Potential involvement of kinesin-1 in the regulation of subcellular localization of Girdin (2015)
  • Author(s): Aya Muramatsu, Atsushi Enomoto, Takuya Kato, Liang Weng, Keisuke Kuroda, Naoya Asai, Masato Asai, Shinji Mii, Masahide Takahashia
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 463 Issue: 4 Pages: 999-1005
  • DOI: 10.1016/j.bbrc.2015.06.049
  • Peer Reviewed / Open Access
• [Presentation] KANPHOS (Kinase-Associated Phospho-Signaling) Platform - A comprehensive database for kinase-associated neural phosphorylation signaling (2019)
  • Author(s): Keisuke Kuroda, Taku Nagai, Mutsuki Amano, Junichiro Yoshimoto, Takayuki Kannon, Tomoki Nishioka, Shiro Usui, Kozo Kaibuchi
  • Organizer: 第92回薬理学会年会
• [Presentation] What kind of intracellular signals are controlled by the antipsychotics in the brain? (2018)
  • Author(s): Keisuke Kuroda
  • Organizer: 第40回日本生物学的精神医学会・第61回日本神経化学会大会 合同年会
  • Invited
• [Presentation] KANPHOS (Kinase-Associated Phospho-Signaling) Platform - 新規リン酸化DBプラットフォーム - (2017)
  • Author(s): 黒田 啓介, 永井 拓, 天野 睦紀, 吉本 潤一郎, 観音 隆幸, 西岡 朋生, 臼井 支朗, 貝淵 弘三
  • Organizer: 第90回日本薬理学会年会
  • Place of Presentation: 長崎ブリックホール、長崎新聞文化ホール（長崎県長崎市）
  • Year and Date: 2017-03-15
• [Presentation] RhoAはスパイン形態と行動を制御する (2017)
  • Author(s): 中ノ瀬 友稀, 黒田 啓介, 張 心健, 織田 海秀, 永井 拓, 貝淵 弘三
  • Organizer: 第90回日本薬理学会年会
  • Place of Presentation: 長崎ブリックホール、長崎新聞文化ホール（長崎県長崎市）
  • Year and Date: 2017-03-15
• [Presentation] 14－3－3を用いた細胞種特異的リン酸化タンパク質の単離 (2017)
  • Author(s): 吉川 麻里奈, 黒田 啓介, 永井 拓, 貝淵 弘三
  • Organizer: 第90回日本薬理学会年会
  • Place of Presentation: 長崎ブリックホール、長崎新聞文化ホール（長崎県長崎市）
  • Year and Date: 2017-03-15
• [Presentation] RhoA activity regulates spine morphology and memory associated behavior. (2017)
  • Author(s): K. Kuroda, X. Zhang, K. Oda, Y. Nakanose, M. Yoshikawa, T. Nagai, K. Kaibuchi
  • Organizer: ASCB | EMBO 2017 Meeting
  • Int'l Joint Research
• [Presentation] Cell‐type‐specific isolation of 14‐3‐3 associated phosphoprotein from complex brain tissues. (2017)
  • Author(s): M. Yoshikawa, K. Kuroda, T. Nagai, K. Kaibuchi
  • Organizer: ASCB | EMBO 2017 Meeting
  • Int'l Joint Research
• [Presentation] 精神疾患の分子メカニズム解明への挑戦～ドーパミンの細胞内シグナル伝達機構解析について～ (2016)
  • Author(s): 黒田 啓介
  • Organizer: 第３８回日本生物学的精神医学会大会・第５９回日本神経化学会大会 合同年会
  • Place of Presentation: 福岡国際会議場（福岡県福岡市）
  • Year and Date: 2016-09-08
• [Presentation] Analysis of effect of RhoA activity on spine morphology and memory formation (2016)
  • Author(s): 黒田啓介、張心健、織田海秀、永井拓、貝淵弘三
  • Organizer: 第89回日本薬理学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-03-09
• [Remarks] 大規模リン酸化プロテオミクス解析で快感を生み出すメカニズムを解明
  • URL: http://www.med.nagoya-u.ac.jp/medical/dbps_data/_material_/nu_medical/_res/topix/2015/Rap1_20160122jp.pdf