Cloning, expression and immunolocalization of alpha(1)-adrenoceptor in different tissues from rhesus monkey and human male reproductive tract
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2008-02-01
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Patrão, Marilia Tavares Coutinho da Costa [UNIFESP]
Queiroz, Daniel Barboza Cava [UNIFESP]
Grossman, Gail
Petrusz, Peter
Lazari, Maria de Fatima Magalhaes [UNIFESP]
Avellar, Maria Christina Werneck [UNIFESP]
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This study reports the genomic organization of the rhesus alpha(1A)-adrenoceptor gene (ADRA1A). Full-length cloning of rhesus ADRA1A splice variants was achieved by combining PCR screening of a seminal vesicle cDNA library and 5'-RACE assays with total RNA from seminal vesicle. the classical ADRA1A mRNA (ADRA1A_nu 1) and six full-length ADRA1A splice variants were identified representing transcripts that code for functional (ADRA1A_nu 1, ADRA1A_nu 2a, ADRA1A_nu 3a, ADRA1A_nu 3d, ADRA1A_nu 3e) and truncated (ADRA1A_nu 2c and ADRA1A_nu 3c) receptor isoforms. Comparative analysis of the deduced amino acid sequence indicated that rhesus ADRA1A_i1 isoform (corresponding to the ADRA1A_nu 1 transcript) shares high identity to the amino acid sequence present in the classical alpha(1A)-adrenoceptor from human and other mammalian species. Partial nucleotide sequences for rhesus alpha(1B)-(ADRA1B) and alpha(1D)-adrenoceptor (ADRA1D) transcripts were also characterized. RT-PCR studies indicated differential distribution of all ADRA1A-related splice variants as well as ADRA1B and ADRA1D mRNAs, in tissues from rhesus and human male reproductive tract. Immunohistochemistry revealed alpha(1A)-adrenoceptor (ADRA1A_ i1) immunostaining in smooth muscle cells and epithelial cells of rhesus efferent ductules, epididymis and seminal vesicle. Taken together the present results demonstrate that the complexity of the splicing mechanisms involved in the regulation of the ADRA1A gene is not restricted to human and is a common characteristic among Old World monkeys.
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Molecular Human Reproduction. Oxford: Oxford Univ Press, v. 14, n. 2, p. 85-96, 2008.