Normal electrophysiological and behavioral responses to ethanol in mice lacking the long splice variant of the gamma2 subunit of the gamma-aminobutyrate type A receptor.
The gamma subunit of the gamma-aminobutyric acid type A receptor (GABA(A)-R) is essential for bestowing both normal single channel conductance and sensitivity to benzodiazepines on native GABA(A)-Rs. The long splice variant of the gamma2 subunit (gamma2L) has been postulated to be essential in mediating the modulatory actions of ethanol at the GABA(A)-R. In order to evaluate this hypothesis, gene targeting was used to delete the 24bp exon which distinguishes gamma2L from the short splice variant (gamma2S). Mice homozygous for this exon deletion (gamma2L-/-) are viable and indistinguishable from wild-type (gamma2L+/+) mice. No gamma2L mRNA was detected in these mice, nor could gamma2L-containing GABA(A)-R protein be detected by specific antibodies. Radioligand binding studies showed the total amount of gamma2 subunit protein to be not significantly changed, suggesting that gamma2S replaces gamma2L in the brains of the knockout animals. Electrophysiological recordings from dorsal root ganglion neurons revealed a normal complement of functional receptors. There was no difference in the potentiation of GABA currents by ethanol (20-200 mM) observed in neurons from gamma2L+/+ or gamma2L-/- mice. Several behavioral effects of ethanol, such as sleep time, anxiolysis, acute functional tolerance, chronic withdrawal hyperexcitability and hyperlocomotor activity were also unaffected by genotype. It is concluded that gamma2L is not required for ethanol's modulatory action at the GABA(A)-R or whole animal behavioral effects.
Homanics
G E
GE
Department of Anesthesiology/Critical Care Medicine, University of Pittsburgh School of Medicine, PA 15261, USA. homanics@smtp.anes.upmc.edu
Harrison
N L
NL
Quinlan
J J
JJ
Krasowski
M D
MD
Rick
C E
CE
de Blas
A L
AL
Mehta
A K
AK
Kist
F
F
Mihalek
R M
RM
Aul
J J
JJ
Firestone
L L
LL
D017398:Alternative Splicing; D000818:Animals; D001007:Anxiety; D001921:Brain / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D002678:Chimera; D003433:Crosses, Genetic; D000431:Ethanol / Q000494:pharmacology*; D005091:Exons; D005260:Female; D005445:Flunitrazepam / Q000493:pharmacokinetics; D005727:Ganglia, Spinal / Q000502:physiology*; D014644:Genetic Variation; D046911:Macromolecular Substances; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009043:Motor Activity / Q000187:drug effects; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D011869:Radioligand Assay; D011963:Receptors, GABA-A / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D017384:Sequence Deletion; D012890:Sleep / Q000187:drug effects; D013375:Substance Withdrawal Syndrome; D014158:Transcription, Genetic
D046911:Macromolecular Substances; D011963:Receptors, GABA-A; D000431:Ethanol; D005445:Flunitrazepam