Ablation of glutamate receptor GluD2 in adult Purkinje cells causes multiple innervation of climbing fibers by ectopic innervation of transverse collaterals

Content Provider	Semantic Scholar
Author	Miyazaki, Taisuke Yamasaki, Miwako Takeuchi, Tomonori Sakimura, Kenji Watanabe, Masahiko
Copyright Year	2010
Abstract	Trafficking of AMPA-type glutamate receptor (AMPAR) around the postsynaptic membrane is crucial for normal synaptic function and plasticity, and the accumulation of postsynaptic AMPAR occurs in the hippocampal longterm potentiation (LTP). There are two routes for the postsynaptic delivery of AMPAR: lateral diffusion on the cell membrane and exocytosis from the intracellular vesicles. However, the detailed process is elusive. To address this issue, we have developed a novel experimental method to live-image the location and movement of AMPAR with a high signal-to-noise ratio using TIRFM (Total Internal Reflection Fluorescence Microscopy). The cover glass was coated with Neurexin, which is a type of presynaptic adhesion molecule binding to postsynaptic Neuroligin and contributing to synapse formation. Then, rat hippocampal neurons were cultured on the glass. A postsynaptic marker protein PSD-95 was detected just on the glass in the culture, suggesting that the postsynaptic membrane was formed directly on the glass. We next expressed an AMPAR subunit GluA1 fused to SEP (Super Ecliptic pHluorin, a pH-sensitive fluorescent protein) in cultured hippocampal neurons, and observed the GluA1 movement around the postsynaptic membrane by TIRFM. Both exocytosis and lateral diffusion of GluA1 were recorded. The electrical tetanic stimulation increased the occurrence of exocytosis around the postsynaptic membrane, suggesting the implication of GluA1 exocytosis in LTP.
File Format	PDF HTM / HTML
DOI	10.1016/j.neures.2010.07.146
Volume Number	68
Alternate Webpage(s)	https://api.elsevier.com/content/article/pii/S0168010210003172
Alternate Webpage(s)	https://www.sciencedirect.com/science/article/pii/S0168010210003172?dgcid=api_sd_search-api-endpoint
Alternate Webpage(s)	https://doi.org/10.1016/j.neures.2010.07.146
Journal	Neuroscience Research
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article