Drs. Casalotti, Mamalaki, and Barnard is MRC Molecular Neurobiology Unit, MRC Centre, Hills Road, Cambridge CB2 2QH, U.K.
Antibodies Recognising the GABAA/Benzodiazepine Receptor Including Its Regulatory Sites
Article first published online: 12 DEC 2006
Journal of Neurochemistry
Volume 46, Issue 3, pages 854–861, March 1986
How to Cite
Stephenson, F. A., Casalotti, S. O., Mamalaki, C. and Barnard, E. A. (1986), Antibodies Recognising the GABAA/Benzodiazepine Receptor Including Its Regulatory Sites. Journal of Neurochemistry, 46: 854–861. doi: 10.1111/j.1471-4159.1986.tb13050.x
- Issue published online: 12 DEC 2006
- Article first published online: 12 DEC 2006
- Received June 24, 1985; accepted September 24, 1985
Abstract: Polyclonal antibodies have been raised against the GABAA/benzodiazepine receptor purified to homogeneity from bovine cerebral cortex in deoxycholate and Triton X-100 media. Radioimmunoassay was applied to measure specific antibody production using the 125I-la-belled -γ-aminobutyric acid (GABA)/benzodiazepine receptor as antigen. The antibodies specifically immuno-precipitated the binding sites for [3H]muscimol and for [3H]flunitrazepam from purified preparations. In addition, when a 3-[(3-cholamidopropyl)dimethylammonio] 1-propanesulphonate (CHAPS) extract of bovine brain membranes was treated with the antibodies, those sites as well as the [3H]propyl-ß-carboline-3-carboxylate binding, the [35S]t-butylbicyclophosphorothionate binding (TBPS), the barbiturate-enhanced [3H]fluni-trazepam binding, and the GABA-enhanced [3H]fluni-trazepam binding were all removed together into the immunoprecipitate. Western blot experiments showed that these antibodies recognise the α-subunit of the purified GABA/benzodiazepine receptor. These results further support the existence in the brain of a single protein, the GABAA receptor, containing a set of regulatory binding sites for benzodiazepines and chloride channel modulators.