Assembly of functional GABAA receptors in insect cells using baculovirus expression vectors.
We have constructed recombinant baculoviruses containing cDNAs encoding either the alpha 1- or the beta 1-subunit of the bovine GABAA receptor. In Spodoptera frugiperda (IPLB-Sf-21) cells infected with recombinant virus expressing either the alpha 1- or beta 1-subunit, or in cells co-infected with both viruses, functional GABAA receptors were detected by whole-cell electrophysiological recordings. The threshold for the responses mediated by the homo-oligomeric channels (alpha- or beta-) was 2-3 x 10(-6) M GABA, and for the co-infected cells was 8 x 10(-8) M GABA, suggesting that hetero-oligomeric channels formed in these cells. All GABA-induced currents were found to be inhibited by bicuculline and picrotoxin, potentiated by pentobarbital but were insensitive to benzodiazepines.
Atkinson
A E
AE
School of Biological and Molecular Sciences, Oxford Polytechnic, Headington, UK.
Bermudez
I
I
Darlison
M G
MG
Barnard
E A
EA
Earley
F G
FG
Possee
R D
RD
Beadle
D J
DJ
King
L A
LA
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001640:Bicuculline / Q000494:pharmacology; D002417:Cattle; D002460:Cell Line; D003001:Cloning, Molecular; D005071:Evoked Potentials / Q000187:drug effects; D005822:Genetic Vectors; D046911:Macromolecular Substances; D009036:Moths; D010424:Pentobarbital / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D014162:Transfection; D010566:Virulence Factors, Bordetella / Q000494:pharmacology; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology*
D046911:Macromolecular Substances; D011963:Receptors, GABA-A; D011994:Recombinant Proteins; D010566:Virulence Factors, Bordetella; D005680:gamma-Aminobutyric Acid; D010424:Pentobarbital; D001640:Bicuculline