Immunohistochemical analysis of neuronal networks in brain using antibody for neurotransmitter receptors

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Immunohistochemical procedures for cerebral GABA_A and GABA_B receptors using specific monoclonal antibodies have been developed. The results obtained are as follow. 1)Albin o rabbits were immunized against partially purified GABA_A receptor complex from the bovine brain using baclofen-affinity column, and the specific antibody for GABA_A receptor complex was obtained. 2) Mice were immunized against partially purified GABA_B receptor protein using baclofen-affinity column, and the monoclonal antibody which recognizes 80KDa GABA_B receptor was obtained. 3) The immunoaffinity column using GABA_B receptor monoclonal antibody was obtained, and complete purification of the GABA_B receptor protein(80KDa) was achieved using this immunoaffinity column. 4) The purified GABA_B receptor possessed pharmacological properties as GABA_B receptor and exhibited the inhibition of cAMP formation when it was reconstituted in phospholipid vesicles with purified adenylate cyclase and Gi/Go type of GTP binding protein. 5) Immunohistochemical studies using above GABA_A receptor and GABA_B receptor antibodies revealed that the localization of GABA_A receptor and GABA_B receptors at GABAergic synapses was different. 6) It has been confirmed that these immunohistochemical procedures are useful for the analysis of GABAergic pathways in the brain.