The GABAA/benzodiazepine receptor complex in rat brain neuronal cultures. Characterization by immunoprecipitation.
The expression of the GABAA/benzodiazepine receptor (GABAR/BZDR) complex in primary neuronal cultures from rat brain embryos has been investigated. The GABAR/BZDR complex was photoaffinity labeled with [3H]flunitrazepam [3H]FNZ and immunoprecipitated with subunit specific antibodies. These were the mAb 62-3G1 which is specific for the 57-kDa GABA binding subunit, and the rabbit antiserum A which recognizes the 51-kDa [3H]FNZ binding subunit. The results indicate that the cultured neurons express 5 different peptides of 51, 53, 54, 57 and 59 kDa that can be photoaffinity labeled with [3H]FNZ and that all of them are physically coupled to the GABAA receptor. Most of the [3H]FNZ photolabeled peptides have similar mobilities to those found in the brain of the newborn rat. Nevertheless, some of the quantitative changes in the photolabeled peptides observed during the normal development of the rat brain were not observed or occurred at much slower pace in the cultured neurons.
Vitorica
J
J
Division of Molecular Biology and Biochemistry, School of Basic Life Sciences, University of Missouri-Kansas City 64110-2499.
Park
D
D
de Blas
A L
AL
D000345:Affinity Labels; D000818:Animals; D000831:Animals, Newborn / Q000378:metabolism; D001921:Brain / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D002417:Cattle; D002478:Cells, Cultured; D005260:Female; D005445:Flunitrazepam; D066298:In Vitro Techniques; D009474:Neurons / Q000378:metabolism*; D011233:Precipitin Tests; D011247:Pregnancy; D051381:Rats; D011963:Receptors, GABA-A / Q000276:immunology / Q000378:metabolism*
D000345:Affinity Labels; D011963:Receptors, GABA-A; D005445:Flunitrazepam