Measuring dynamics of synaptic glutamate receptors

2014 Fiscal Year Final Research Report

• Project/Area Number: 26560457
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Basic / Social brain science
• Research Institution: The University of Tokyo
• Principal Investigator: BITO Haruhiko 東京大学, 医学(系)研究科(研究院), 教授 (00291964)
• Project Period (FY): 2014-04-01 – 2015-03-31
• Keywords: 脳神経 / 神経科学 / シグナル伝達

Outline of Final Research Achievements

In this study, we aimed to develop a sensitive real-time imaging method which takes advantage of recently introduced wash-free chemical ligands to probe the lateral diffusion and exo-endocytosis of glutamate receptors during synaptic plasticity in single spines. AMPA receptor subunits in which enzyme tags were introduced were expressed were transfected into primary hippocampal neurons in culture, and an expression protocol was determined to express the exogenous surface receptors to a degree comparable to endogenous synaptic glutamate receptors. Using screened small molecular weight chemicals, we further developed a versatile surface receptor imaging protocol. Finally, we identified a useful chemical ligand that enabled pulse-chase labeling and imaging of newly membrane-inserted receptors. Preliminary experiments were carried out to test these imaging tools and to measure real-time GluA1 dynamics in neurons undergoing plasticity.

Free Research Field

神経生化学