Premium
Virus‐mediated swapping of zolpidem‐insensitive with zolpidem‐sensitive GABA A receptors in cortical pyramidal cells
Author(s) -
Sumegi Mate,
Fukazawa Yugo,
Matsui Ko,
Lorincz Andrea,
Eyre Mark D.,
Nusser Zoltan,
Shigemoto Ryuichi
Publication year - 2012
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2012.227538
Subject(s) - zolpidem , gabaa receptor , receptor , immunogold labelling , patch clamp , protein subunit , microbiology and biotechnology , chemistry , in vitro , wild type , biology , biophysics , transgene , biochemistry , pharmacology , gene , anatomy , mutant , ultrastructure , insomnia
Key points • We generated lenti‐ and adeno‐associated viruses which were used to replace the zolpidem‐insensitive GABA A receptors of a transgenic mouse line with wild‐type, zolpidem‐sensitive ones. • The virally expressed wild‐type, zolpidem‐sensitive GABA A receptor γ2 subunits were tagged with a small immunotag (AU1). • Light microscopic fluorescent and electron microscopic freeze‐fracture immunogold labelling revealed that the virally introduced AU1‐tagged γ2 subunit‐containing receptors had a normal synaptic distribution on cortical pyramidal cells. • In vitro patch‐clamp recordings demonstrated that the insertion of this immunotag did not alter the kinetic and pharmacological properties of the virally inserted γ2 subunits. • Our results demonstrate a novel transgenic–viral pharmacogenetic approach, which allows the selective silencing of well‐defined neuronal populations in the brain.