The overall objective of this research project is to elucidate the chemical structure and the mode of action of endogenous ligands for benzodiazepine (BZ)/GABA receptors with a special focus on their role in the regulation of GABAergic transmission. More specifically, the plan is to perform the following studies: 1. Endogenous ligands for benzodiazepine receptor(s) (BZR) will be purified so that sufficient quantity of the purified endogenous ligands can be obtained for functional analysis and structural determination. 2. Ligand binding studies and behavioral tests will be conducted to determine the specificity and the mode of action of endogenous ligands, e.g. agonist, antagonist, of inverse agonist of the central type BZR. In addition, the effect of endogenous ligands on the GABA binding site, barbiturates binding site, and the Cl.. ion channel will also be investigated. 3. Purified endogenous ligands as haptens will be used and conjugated to a carrier protein (e.g. bovine serum albumin or hemocyanin) to serve as antigen for the production of polyclonal and monoclonal antibodies. These studies are important for understanding the ligand binding characteristics and competition for receptor sites on nerve cell membranes.
