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Sindbis viral‐mediated expression of Ca 2+ ‐permeable AMPA receptors at hippocampal CA1 synapses and induction of NMDA receptor‐independent long‐term potentiation
Author(s) -
Okada Takashi,
Yamada Nobuaki,
Kakegawa Wataru,
Tsuzuki Keisuke,
Kawamura Meiko,
Nawa Hiroyuki,
Iino Masae,
Ozawa Seiji
Publication year - 2001
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1046/j.0953-816x.2001.01523.x
Subject(s) - ampa receptor , long term potentiation , glutamate receptor , microbiology and biotechnology , biology , neuroscience , excitatory postsynaptic potential , neurotransmission , sindbis virus , receptor , chemistry , inhibitory postsynaptic potential , biochemistry , gene , rna
Abstract Gene manipulation in order to artificially express a particular gene in neurons in the central nervous system is a powerful tool for the analysis of brain function. Sindbis viral vectors have been developed to express high levels of foreign genes in postmitotic brain neurons with little transfection of glial cells. In this study, we expressed the gene encoding the unedited GluR2 (GluR‐B) subunit of the AMPA‐type glutamate receptor that forms inwardly rectifying and Ca 2+ ‐permeable channels, in rat CA1 hippocampal neurons in slice cultures using Sindbis viral vectors. The pyramidal cell layer of the CA1 region was injected with recombinant Sindbis viruses encoding both enhanced green fluorescent protein (GFP) and unedited GluR2. The GFP fluorescence from CA1 neurons could be detected as early as 6 h and reached a maximal level about 48 h postinfection. The inwardly rectifying and Ca 2+ ‐permeable AMPA receptors were expressed in most CA1 pyramidal cells expressing GFP. These AMPA receptors expressed by gene transfer were involved in fast excitatory neurotransmission elicited by electrical stimulation of the Schaffer collaterals in the stratum radiatum. Tetanic stimulation of Schaffer collaterals induced NMDA receptor‐independent, long‐term potentiation due to Ca 2+ influx through the newly expressed AMPA receptors in the area densely stained with GFP. Thus, the combined use of Sindbis viral vectors with the GFP reporter allowed physiological examination of the roles of a specific gene product in synaptic function in well‐characterized brain neurons.