[unreadable] [unreadable] Gene therapy techniques need substantial development to provide therapeutic possibilities for treating neurological disorders such as Parkinson's disease (PD). Based on molecular control mechanism of noradrenergic neuron-specific gene transcription, we recently devised a gene delivery system that can efficiently target transgene expression to noradrenergic neurons in a cell-specific manner. Our long-term goal is to establish gene therapy system(s) that will drive efficient transgene expression in a dopamine (DA) neuron-specific fashion based on discovery and characterization of DA-specific genes. Toward this end, we propose to identify and isolate genes that are selectively expressed in DA neurons by analyzing the gene expression profiles of the mid-brain area using the most comprehensive cDNA microar-rays such as the augmented NIA 16K chip and augmented RIKEN 16 K chip Because these chips do not cover the whole genome yet, we will identify novel DA-specific genes by the PCR-based subtractive hybridization techniques. Expression patterns of putative DA-specific genes will be tested by semiquantitative RT-PCR and confirmed by in situ hybridization. [unreadable] [unreadable] Among the isolated DA-specific genes, we will first focus on putative DNA-binding transcription factors The consensus binding sites for these putative transcription factors will be defined and their potential promoter function will be tested by cotransfection assays using cell line systems. On the basis of the mechanism of action of the novel DA-specific transcription factor(s), synthetic promoters will be developed and optimized. The optimized synthetic promoter will be subcloned in front of the reporter lacZ gene in the context of the self-inactivated Lenti viral vectors. Cell type-specific expression of the reporter gene will be examined using both in vitro mesencephalic primary neuronal cultures as well as in different rat brain areas following stereotactic injection. At the later stage of this proposal, we plan to use our developed promoter system(s) to deliver therapeutic genes (e.g., GDNF and antioxidants) to the DA neurons and will test whether they can efficiently ameliorate behavioral symptoms in animal models of PD. [unreadable] [unreadable] The proposed research will identify and isolate genes that are selectively expressed in DA neurons on a genome-wide scale and will characterize their transcriptional regulatory mechanisms. Based on these mechanisms, we will devise novel DA neuron-specific promoter systems and test them using in vitro and in vivo systems. In combination with safe viral vector systems, our developed gene delivery systems can be translated clinically into gene therapy approaches for PD and other neurological disorders, in which DA neuronal system is dysregulated.