GABAA receptor subtypes immunopurified from rat brain with alpha subunit-specific antibodies have unique pharmacological properties.
The unique cytoplasmic loop regions of the alpha 1, alpha 2, alpha 3, and alpha 5 subunits of the GABAA receptor were expressed in bacterial and used to produce subunit-specific polyclonal antisera. Antibodies immobilized on protein A-Sepharose were used to isolate naturally occurring alpha-specific populations of GABAA receptors from rat brain that retained the ability to bind [3H]muscimol, [3H]flunitrazepam, [3H]Ro15-1788, and [125I]iodo-clonazepam with high affinity. Pharmacological characterization of these subtypes revealed marked differences between the isolated receptor populations and was generally in agreement with the reported pharmacological profiles of GABAA receptors in cells transiently transfected with alpha 1 beta 1 gamma 2, alpha 2 beta 1 gamma 2, alpha 3 beta 1 gamma 2, and alpha 5 beta 1 gamma 2 combinations of subunits. Additional subtypes were also identified that bind [3H]muscimol but do not bind benzodiazepines with high affinity. The majority of GABAA receptor oligomers contains only a single type of alpha subunit, and we conclude that alpha 1, alpha 2, alpha 3, and alpha 5 subunits exist in vivo in combination with the beta subunit and gamma 2 subunit.
McKernan
R M
RM
Department of Biochemistry, Merck Sharp and Dohme Research Laboratories, Harlow, Essex, England.
Quirk
K
K
Prince
R
R
Cox
P A
PA
Gillard
N P
NP
Ragan
C I
CI
Whiting
P
P
D000818:Animals; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D010455:Peptides / Q000276:immunology; D011233:Precipitin Tests; D051381:Rats; D011963:Receptors, GABA-A / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
D000906:Antibodies; D010455:Peptides; D011963:Receptors, GABA-A