Elucidating the mechanisms underlying selective NR3A accumulation at non-synaptic junctions

• Project/Area Number: 17KK0160
• Research Category: Fund for the Promotion of Joint International Research (Fostering Joint International Research)
• Allocation Type: Multi-year Fund
• Research Field: General anatomy (including histology/embryology)
• Research Institution: Hokkaido University
• Principal Investigator: Yamasaki Miwako 北海道大学, 医学研究院, 准教授 (10431305)
• Project Period (FY): 2017 – 2019
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥14,430,000 (Direct Cost: ¥11,100,000、Indirect Cost: ¥3,330,000)
• Keywords: NMDA受容体 / 抑制性ニューロン / シナプス / 小脳 / グルタミン酸受容体

Outline of Final Research Achievements

In the preceding project, we found that NR3A selectively localized to non-synaptic contacts between climbing fiber and cerebellar stellate cells, and those between glutamatergic terminals and somatostatin-positive GABAergic interneurons in the cerebral cortex. To identify molecular partners that constitute biochemical complex, we immunopurified biochemical complex containing NR3A and conducted analysis by mass spectrometry. We found that NR1 but not Kv4.3 was included in the complex. Furthermore, in a reconstitution system using Xenopus oocytes, NR3A assembly required NR1, but not Kv4.3. Rescue experiment using AAV revealed that NR1 was required for selective NR3A expression at non-synaptic contacts.

Academic Significance and Societal Importance of the Research Achievements

脳機能を担う神経回路の発達においてNMDA型グルタミン酸受容体NR3Aは重要な役割を持つが、発現部位やその機能に不明な点が多く解明が期待されている。本研究では、組織学的解析により見出した「NR3A受容体と共局在する分子」の生化学的スクリーニングを行い、NR3A受容体の特徴である「特定の神経回路におけるユニークな集積パターン」に必要な分子とそうでない分子を明らかにした。こうした知見は今後の脳回路研究の発展に貢献することが期待される。

Research Products

• [Int'l Joint Research] Yale大学(米国) (2018)
  • Year and Date: 2018-03-23
• [Journal Article] Expression mapping, quantification, and complex formation of GluD1 and GluD2 glutamate receptors in adult mouse brain (2019)
  • Author(s): Nakamoto Chihiro、Konno Kohtarou、Miyazaki Taisuke、Nakatsukasa Ena、Natsume Rie、Abe Manabu、Kawamura Meiko、Fukazawa Yugo、Shigemoto Ryuichi、Yamasaki Miwako、Sakimura Kenji、Watanabe Masahiko
  • Journal Title: Journal of Comparative Neurology Volume: 528 Issue: 6 Pages: 1003-1027
  • DOI: 10.1002/cne.24792
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] mGluR1 in cerebellar Purkinje cells is essential for the formation but not expression of associative eyeblink memory (2019)
  • Author(s): Nakao Harumi、Kishimoto Yasushi、Hashimoto Kouichi、Kitamura Kazuo、Yamasaki Miwako、Nakao Kazuki、Watanabe Masahiko、Kano Masanobu、Kirino Yutaka、Aiba Atsu
  • Journal Title: Scientific Reports Volume: 9 Issue: 1 Pages: 7353-7353
  • DOI: 10.1038/s41598-019-43744-z
  • Peer Reviewed / Open Access
• [Journal Article] Localization of phospholipase C β3 in the major salivary glands of adult mice. (2019)
  • Author(s): Rawangwong A, Pidsaya A, Thoungseabyoun W, Tachow A, Sawatpanich T, Sakaew W, Yamasaki M, Watanabe M, Kondo H, Hipkaeo W.
  • Journal Title: Acta Histochem. Volume: 121 Issue: 4 Pages: 484-490
  • DOI: 10.1016/j.acthis.2019.04.006
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Heterogeneous localization of muscarinic cholinoceptor M1 in the salivary ducts of adult mice. (2019)
  • Author(s): Rawangwong A, Khrongyut S, Chomphoo S, Konno K, Yamasaki M, Watanabe M, Kondo H, Hipkaeo W.
  • Journal Title: Arch Oral Biol. Volume: 100 Pages: 14-22
  • DOI: 10.1016/j.archoralbio.2019.02.001
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Silent Learning. (2018)
  • Author(s): Rossato JI, Moreno A, Genzel L, Yamasaki M, Takeuchi T, Canals S, Morris RGM.
  • Journal Title: Curr Biol. Volume: 28 Issue: 21 Pages: 3508-3515
  • DOI: 10.1016/j.cub.2018.09.012
  • Peer Reviewed / Int'l Joint Research
• [Presentation] Distinct central synapse that shows unique property to maintain AMPAR numbers equal (2018)
  • Author(s): Miwako Yamasaki
  • Organizer: Gordon Research Conference
  • Int'l Joint Research