Abstract

5-hydroxytryptamine (5-HT, serotonin) and its receptors have been implicated in the etiology and therapeutic treatment of human mental depression. A basic knowledge of 5-HT receptors will help to better understand the molecular mechanism of 5-HT mediated neural function and will provide insights into the molecular basis of mental depression. The objectives of this project are to investigate the structure and function relationships, the genomic organization and the regulation of human 5-HT-2 receptor(s). These objectives will be accomplished by using the rat brain 5-HT-2 receptor cDNA cloned recently.
The specific aims of the proposed study are outlined below. 1. To clone 5-HT-2 receptor and related cDNAs from human brain. Various 5-HT-2 receptor subtypes and related cDNA clones will be isolated and sequenced. The pharmacological profiles and the second messenger systems linked to the expressed receptors (phosphoinositol (PI) turnover, CA++ release, or adenylate cyclase) will be characterized. 2. To map the chromosomal location of the 5-HT-2 receptor gene.
This aim will be accomplished by somatic cell mapping and in situ hybridization of the chromosome. 3. To isolate human genomic 5-HT-2 receptor clones. 4. To construct the restriction map and to sequence human 5-HT-2 receptor genomic clones. 5. To identify and to analyze promoter regions of this gene.
This aim will be accomplished by RNase protection, primer extension, and chloramphenicol acetyltransferase (CAT) reporter gene assay. The cis- elements, trans-acting factors and the functions of these elements will be systematically analyzed. 6a. To investigate if 5-HT-2 receptor gene is regulated by protein kinase C (PKC) and/or protein kinase A (PKA). The effects of TPA (12-0- tetradecanoyl-phorbol-14 acetate, activator of PKC) on the 5-HT-2 receptor gene expression will be studied by using CAT report gene assay and footprinting analysis. The ability of 1-(5-isoquinolylsulfonyl)-2- methylpiperazine (H7) to block TPA activation will be studied. Similar studies will be performed using activators of PKA such as forskolin and 8- bromodibutyryl cAMP. 6b. To investigate if antagonist induced 5-HT-2 receptor down regulation is at the transcriptional level.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Research Project (R01)
Project #
5R01MH037020-08
Application #
3375999
Study Section
Neurosciences Research Review Committee (BPN)
Project Start
1991-03-01
Project End
1996-02-29
Budget Start
1992-03-01
Budget End
1993-02-28
Support Year
8
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
University of Southern California
Department
Type
Schools of Pharmacy
DUNS #
041544081
City
Los Angeles
State
CA
Country
United States
Zip Code
90089

  Publications

Wang, Tso-Jen; Huang, San-Yuan; Lin, Wei-Wen et al. (2007) Possible interaction between MAOA and DRD2 genes associated with antisocial alcoholism among Han Chinese men in Taiwan. Prog Neuropsychopharmacol Biol Psychiatry 31:108-14
Shih, J C; Chen, K (2004) Regulation of MAO-A and MAO-B gene expression. Curr Med Chem 11:1995-2005
Yeung Lam, Philip; Chen, Kevin; Shih, Jean C (2004) The circadian rhythm of 5-HT biosynthetic and degradative enzymes in immortalized mouse neuroendocrine pineal cell line--a model for studying circadian rhythm. Life Sci 75:3017-26
Ou, Xiao-Ming; Chen, Kevin; Shih, Jean C (2004) Dual functions of transcription factors, transforming growth factor-beta-inducible early gene (TIEG)2 and Sp3, are mediated by CACCC element and Sp1 sites of human monoamine oxidase (MAO) B gene. J Biol Chem 279:21021-8
Shih, Jean Chen (2004) Cloning, after cloning, knock-out mice, and physiological functions of MAO A and B. Neurotoxicology 25:21-30
Lu, Ru-Band; Lin, Wei-Wen; Lee, Jia-Fu et al. (2003) Neither antisocial personality disorder nor antisocial alcoholism is associated with the MAO-A gene in Han Chinese males. Alcohol Clin Exp Res 27:889-93
Vitalis, Tania; Alvarez, Chantal; Chen, Kevin et al. (2003) Developmental expression pattern of monoamine oxidases in sensory organs and neural crest derivatives. J Comp Neurol 464:392-403
Wong, Wai K; Chen, Kevin; Shih, Jean C (2003) Decreased methylation and transcription repressor Sp3 up-regulated human monoamine oxidase (MAO) B expression during Caco-2 differentiation. J Biol Chem 278:36227-35
Holschneider, D P; Scremin, O U; Chialvo, D R et al. (2002) Heart rate dynamics in monoamine oxidase-A- and -B-deficient mice. Am J Physiol Heart Circ Physiol 282:H1751-9
Lu, Ru-Band; Lee, Jia-Fu; Ko, Huei-Chen et al. (2002) No association of the MAOA gene with alcoholism among Han Chinese males in Taiwan. Prog Neuropsychopharmacol Biol Psychiatry 26:457-61

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