A mouse model for visualization of GABA(B) receptors.
GABA(B) receptors are the G-protein-coupled receptors for the neurotransmitter gamma-aminobutyric acid (GABA). Receptor subtypes are based on the subunit isoforms GABA(B1a) and GABA(B1b), which combine with GABA(B2) subunits to form heteromeric receptors. Here, we used a modified bacterial artificial chromosome (BAC) containing the GABA(B1) gene to generate transgenic mice expressing GABA(B1a) and GABA(B1b) subunits fused to the enhanced green fluorescence protein (eGFP). We demonstrate that the GABA(B1)-eGFP fusion proteins reproduce the cellular expression patterns of endogenous GABA(B1) proteins in the brain and in peripheral tissue. Crossing the GABA(B1)-eGFP BAC transgene into the GABA(B1) (-/-) background restores pre and postsynaptic GABA(B) functions, showing that the GABA(B1)-eGFP fusion proteins substitute for the lack of endogenous GABA(B1) proteins. Finally, we demonstrate that the GABA(B1)-eGFP fusion proteins replicate the temporal expression patterns of native GABA(B) receptors in cultured neurons. These transgenic mice therefore provide a validated tool for direct visualization of native GABA(B) receptors.
Casanova
Emilio
E
Department of Biomedicine, Institute of Physiology, University of Basel, Basel, Switzerland.
Guetg
Nicole
N
Vigot
Réjan
R
Seddik
Riad
R
Julio-Pieper
Marcela
M
Hyland
Niall P
NP
Cryan
John F
JF
Gassmann
Martin
M
Bettler
Bernhard
B
D000818:Animals; D015153:Blotting, Western; D022202:Chromosomes, Artificial, Bacterial; D017931:DNA Primers / Q000235:genetics; D004594:Electrophysiology; D040461:Gene Components; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D023421:Models, Animal; D018080:Receptors, GABA-B / Q000235:genetics* / Q000378:metabolism*
D017931:DNA Primers; D018080:Receptors, GABA-B; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins