A significant part of native gamma-aminobutyric AcidA receptors containing alpha4 subunits do not contain gamma or delta subunits.
Using a novel antibody directed against the alpha4 subunit of gamma-aminobutyric acidA (GABAA) receptors, 5% of all [3H]muscimol but only about 2% of all [3H]Ro15-4513 binding sites present in brain membrane extracts could be precipitated. This indicated that part of the alpha4 receptors containing [3H]muscimol binding sites did not contain [3H]Ro15-4513 binding sites. Immunoaffinity purification and Western blot analysis of alpha4 receptors demonstrated that not only alpha1, alpha2, alpha3, beta1, beta2, and beta3 subunits but also gamma1, gamma2, gamma3, and delta subunits can be colocalized with alpha4 subunits in native GABAA receptors. Quantification experiments, however, indicated that only 7, 33, 4, or 7% of all alpha4 receptors contained gamma1, gamma2, gamma3, or delta subunits, respectively. These data not only explain the low percentage of [3H]Ro15-4513 binding sites precipitated by the anti-alpha4 antibody but also indicate that approximately 50% of the alpha4 receptors did not contain gamma1, gamma2, gamma3, or delta subunits. These receptors, thus, either are composed of alpha4 and beta1-3 subunits only, or additionally contain epsilon, pi, or so far unidentified GABAA receptor subunits.
Bencsits
E
E
Section of Biochemical Psychiatry, University Clinic for Psychiatry, Währinger Gürtel 18-20, 1090 Vienna, Austria.
Ebert
V
V
Tretter
V
V
Sieghart
W
W
D000911:Antibodies, Monoclonal; D001386:Azides / Q000378:metabolism; D001569:Benzodiazepines / Q000378:metabolism; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D002846:Chromatography, Affinity; D006801:Humans; D009118:Muscimol / Q000378:metabolism; D011233:Precipitin Tests; D011485:Protein Binding; D011963:Receptors, GABA-A / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D001386:Azides; D011963:Receptors, GABA-A; D001569:Benzodiazepines; D009118:Muscimol; C042957:Ro 15-4513