There are major medical reasons for understanding the nature of dopaminergic neurotransmission in the neostriatum. Parkinson's disease,, Huntington's chorea, schizophrenia and attention deficit hyperactivity disorder all reflect disorders of dopamine signaling pathways. This grant application proposes a multi-disciplinary study of signal Project (Biochemical Analysis of Basal Ganglia Phosphoproteins) will include: biochemical characterization of the role of phosphorylation of Th475 and DARPP-32; elucidation of the molecular basis of the interaction of neurabin. Project (Pharmacological Regulation of Basal Ganglia- enriched Phosphoproteins) will include: characterization of the phosphorylation of DARPP-32 at Th473 in intact cells; characterization of the roles of Th475 or Th434 in the regulation of glutamate and GABA receptor phosphorylation using genetically modified mice in which the Th434 and Th475 phosphorylation sites are substituted with alanine; characterization of the role of phosphorylation of spinophilin and neurabin in intact cells; characterization of gene knockout mice to evaluate the roles of PP1 isoforms, and neurabin in the phosphorylation of glutamate and GABA receptors. Project (Physiological and Cell Biological Studies of Phosphoproteins in the Basal Ganglia) will include: electrophysiological analysis of mice lacking either PP1 isoforms, spinophilin, neurabin, or in which the Thr34 and Thr75 phosphorylation sites in DARPP-32 will have been substituted with alanine; characterization of the regional and subcellular localization of spinophilin and neurabin; examination of the mechanism of targeting and the dynamic regulation of the spinophilin-PP1 and neurabin-PP1 complex in neurons. Project (Molecular Biology of Basal Ganglia and Phosphoproteins) will include: analysis of the behavioral and biochemical responses to dopamine in mice lacking striatal PP1 isoforms, mice lacking spinophilin and neurabin, and mice with mutated DARPP-32 genes, in which the Thr34 and Thr 75 phosphorylation sites are substituted with alanine; identification of proteins that interact with spinophilin and neurabin in yeast 2-hybrid systems. Project (Scientific Core) will produce key materials and perform routine required for the other projects.
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