Identification of the GABAA Receptor α3 Subunit in the IMR-32 Neuroblastoma Cell Line |
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Authors: | Penelope J. Noble Shelagh M. P. Anderson Rosemarie J. De Souza Alan J. Cross F. Anne Stephenson |
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Affiliation: | Department of Pharmaceutical Chemistry, School of Pharmacy, London, England;Astra Neuroscience Research Unit, London, England |
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Abstract: | Abstract: A previous report has described the presence of f-[35S]- butylbicyclophosphorothionate binding sites and GABA-gated CI flux in the human neuroblastoma IMR-32 cell line. We now report the further characterisation of this binding site and, even more important, the identification of the GABAA receptor α3 subunit expressed in these cells. Cell membranes prepared from IMR-32 cells were screened by immunoblotting for reactivity with various GABAA receptor a subunit-specific antibodies. Of these, only anti-Cys α3 454-467 antibodies recognised specifically and in a dose-dependent manner an immunoreactive band. This Mr58,000 immunoreactive species and the N -deglycosylated derivatives were both coincident with the respective homologues found in both calf cerebral cortex membranes and purified receptor preparations. This is the first report of the identification of a specific GABAA receptor subunit expressed in a human cell line, and it therefore provides a convenient model for the study of receptor structure and regulation. |
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Keywords: | GABA Benzodiazepine IMR-32 cells Antibody Receptor |
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