Dopamine is an important neurotransmitter in CNS and contributes profoundly to a variety of motor and emotional behaviors. Dopaminergic dysfunction has been associated with several major neuropsychiatric disorders, ranging from drug addiction to schizophrenia to Parkinson's disease. Molecular studies of dopamine function have revealed a set of Dopamine-Regulated Genes (DRGs), which contribute critically to the unique neurochemical and behavioral properties of the striatum. The central hypothesis of this proposal is that these DRGs are co-regulated by a common but complex set of cis-elements and transcription factors. Our primary goal is to identify and validate the clusters of cis-elements for DRG expression (CEDRG) using integrated molecular and bioinformatics approaches.
Specific Aim 1 : We will employ a set of bioinformatics and molecular analysis tools to characterize as fully as possible the genomic organization of the DRGs, with particular effort to determine the TSSs of DRG. We will integrate all genomic information for DRGs into a Web-accessible database, DopamineDB, which will provide a unique resource for investigation of dopamine neurobiology.
Specific Aim 2 : Following phylogenetic analysis of human and mouse DRGs to reveal evolutionally conserved regions within their promoters, we will employ a range of statistical model-based algorithms (Clover [1]) and Glam [2]) to identify statistically over-represented cis-Elements for Dopamine-Regulated Gene expression (CEDRG). We will systematically evaluate the predicted known and novel cis-element binding activity by ChIP-chip analysis and gel shift assay, respectively, in the putative proximal DRG promoters.
Specific Aim 3 : We will determine functional interactions of CEDRGs by detecting statistically significant CEDRG clusters, and by assaying transcription activity of CEDRG clusters in a striatal cloned cell line (ST14A). Furthermore, we will employ a """"""""safe-haven"""""""" transgenic strategy to evaluate in vivo function of identified CEDRG in transgenic mice. Finally, we will examine DRG expression in mice deficient in transcription factors corresponding to the CEDRG to conclusively determine their involvement in DRG expression. The molecular and bioinformatics analyses are integrated throughout the project to overcome major limitations of each individual technique. The information derived from systematic analyses of DRGs will provide critical insights into dopamine functions and identify novel dopamine-regulated transcription factors and thus greatly facilitate the development of novel treatment strategies for dopamine-associated neuropsychiatric disorders such as drug addiction.

Agency
National Institute of Health (NIH)
Institute
National Institute on Drug Abuse (NIDA)
Type
Research Project (R01)
Project #
1R01DA019362-01
Application #
6887605
Study Section
Special Emphasis Panel (ZRG1-IFCN-B (50))
Program Officer
Rutter, Joni
Project Start
2004-07-15
Project End
2009-04-30
Budget Start
2004-07-15
Budget End
2005-04-30
Support Year
1
Fiscal Year
2004
Total Cost
$363,375
Indirect Cost
Name
Boston University
Department
Neurology
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Wei, Catherine J; Augusto, Elisabete; Gomes, Catarina A et al. (2014) Regulation of fear responses by striatal and extrastriatal adenosine A2A receptors in forebrain. Biol Psychiatry 75:855-63
Wei, Catherine J; Singer, Philipp; Coelho, Joana et al. (2011) Selective inactivation of adenosine A(2A) receptors in striatal neurons enhances working memory and reversal learning. Learn Mem 18:459-74
Lazarus, Michael; Shen, Hai-Ying; Cherasse, Yoan et al. (2011) Arousal effect of caffeine depends on adenosine A2A receptors in the shell of the nucleus accumbens. J Neurosci 31:10067-75
Hung, Jui-Hung; Whitfield, Troy W; Yang, Tun-Hsiang et al. (2010) Identification of functional modules that correlate with phenotypic difference: the influence of network topology. Genome Biol 11:R23
Sturgess, Jessica E; Ting-A-Kee, Ryan A; Podbielski, Dominik et al. (2010) Adenosine A1 and A2A receptors are not upstream of caffeine's dopamine D2 receptor-dependent aversive effects and dopamine-independent rewarding effects. Eur J Neurosci 32:143-54
Chen, Jiang-Fan; Yu, Liqun; Shen, Hai-Ying et al. (2010) What knock-out animals tell us about the effects of caffeine. J Alzheimers Dis 20 Suppl 1:S17-24
Boison, D; Chen, J-F; Fredholm, B B (2010) Adenosine signaling and function in glial cells. Cell Death Differ 17:1071-82
Yu, Liqun; Coelho, Joana E; Zhang, Xiaoling et al. (2009) Uncovering multiple molecular targets for caffeine using a drug target validation strategy combining A 2A receptor knockout mice with microarray profiling. Physiol Genomics 37:199-210
Shen, H-Y; Kalda, A; Yu, L et al. (2008) Additive effects of histone deacetylase inhibitors and amphetamine on histone H4 acetylation, cAMP responsive element binding protein phosphorylation and DeltaFosB expression in the striatum and locomotor sensitization in mice. Neuroscience 157:644-55
Yu, Liqun; Haverty, Peter M; Mariani, Juliana et al. (2005) Genetic and pharmacological inactivation of adenosine A2A receptor reveals an Egr-2-mediated transcriptional regulatory network in the mouse striatum. Physiol Genomics 23:89-102