Analysis of the structure of axonal varicosities focusing on active zone proteins

Research Project

Project/Area Number	16K16647
Research Category	Grant-in-Aid for Young Scientists (B)
Allocation Type	Multi-year Fund
Research Field	Basic / Social brain science
Research Institution	University of Yamanashi
Principal Investigator	HAMADA Shun 山梨大学, 大学院総合研究部, 助教 (90755464)
Project Period (FY)	2016-04-01 – 2018-03-31
Project Status	Completed (Fiscal Year 2017)
Budget Amount *help	¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000) Fiscal Year 2017: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000) Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Keywords	バリコシティー / プレシナプス / アクティブゾーン / マイクロ流体デバイス / 軸索分離培養容器 / 脳・神経 / 神経科学 / 細胞・組織 / 生理学
Outline of Final Research Achievements	From morphological analysis of primary cultured hippocampal neurons derived from presynaptic active zone protein CAST, We found that the number and size of varicosities were increased by CAST deletion. Then, we attempted to recover the change of morphology of varicosities in CAST deleted neurons by the infection of the CAST expressing virus. But, there were some unsuitable problems in our experimental environment. To solve these problems, we developed smaller micro fluid device to separate axons from other neuronal cell bodies and dendrites than existing commercial products. By using this device, it was possible to split axons from cell bodies, but difficult to induce the exogenous gene expression. To investigate the efficacy of neurotransmission in individual varicosities, ChR2 was expressed in cultured neurons. Then, we undertook to measure the synaptic responses from non-transfected neurons with stimulating ChR2 expressed axon terminals.