The aim of this proposal is to continue as well as to institute research in the following areas: 1) To study the phosphorylation and activation of tyrosine hydroxylase (TH) and tryptophan hydroxylase (Try-0H) by multiple protein kinases. The role of Ca++/calmodulin multiple protein kinases and of Kinase """"""""C"""""""" in the phosphorylation of TH and Try-0H in specific brain regions and in cultured PC12 cells will be investigated. 2) To develop monoclonal antibodies (m-Ab) to monoamine synthesizing enzymes and to use them as probes for biochemical and immunohistochemical studies. The antibodies will be used to differentiate between various forms of the corresponding enzymes and to distinguish between phospho and dephospho forms of TH and Try-0H. 3) To investigate the mechanisms involved in the neuroleptic-induced activation of TH. The role of various protein kinases in the neuroleptic-induced activation of TH in various brain regions will be studied. (4) To study the recovery rates of DA and Alpha-2 adrenoceptor subtypes after irreversible inactivation with the alkylating agent EEDQ. (5) To study the dopamine (DA) receptors at the molecular level. The solubilized antigens will be used for production of specific monoclonal and/or polyclonal antibodies. (6) To study the dopamine receptors at the molecular level. The solubilized DA antigens will be used for production of specific monoclonal and/or polyclonal antibodies. (7) To study the effects of selective and non-selectived DA agonists on extrapyramidal dysfunctions in monkeys with unilateral ventromedial tegmental lesions of the brain stem and in NMPTP-treated monkeys. (8) To study the DA involvement in a nonhuman primate model which mimics Lesch-Nyhan syndrome.
