| Research Abstract |
Molecular cloning identified three distinct cDNAs encoding alpha_1-adrenoceptors (alpha_<1a>, alpha_<1b>, and alpha_<1d>). Among subtypes, human prostate has been shown to express predominantly alpha_<1a>-adrenoceptors, and these mediate the contractile response. It is already known that two restriction fragment length poylmorphisms (RFLPs) of alpha_<1a>-adrenoceptor gene digested with Pst I restriction enzyme exist in human. This polymorphism might confer an additional putative palmitoylation site in the carboxy-terminal segment of alpha_<1a>-adrenoceptor, implying abnormal signal transduction mechanism. Also, mutation in the third cytoplas-mic loop of alpha_<1a>-adrenoceptor might induce the constitutive activation of the receptor/signal transduc-tion. In the present study, we looked for a possible mutation in the third intracellular loop of the alpha_<1a>-adrenoceptor gene. Fresh prostate tissues were obtained at the time of surgery from 27 patients, frozen in liquid nitrogen, and s
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tored at -80ﾟC.Total RNA was extracted from the prostate tissue, then reverse transcribed, and cDNA was prepared. Fragments of PCR products which encodes the third intracellular loop of alpha_<1a>-adrenoceptor were purified and analysed by DNA sequencer. As a result, no mutation in the third cytoplasmic loop of the alpha_<1a>-adrenoceptor gene was observed. Additionary, we examined whether the Pst I-RFLP of the alpha_<1a>-adrenoceptor is associated with female urethral syndrome. PCR product of 502 bp, which contains the DNA sequence corresponding to Arg492, has been amplified from genomic DNA, and 2 RFLPs of alpha_<1a>-adrenoceptor gene, which are digested with Pst I restriction enzyme into 477bp and 25 bp, were cheked in middle aged women of either control or urethral syndrome. The frequency of alpha_<1a>-adrenoceptor polymorphism was not differently distributed between patients with urethral syndrome and normal subjects in Japan. However, the frequency distribution of this polymorphism was significantly different between normal Japanese and U.S.populations. Our preliminary study does not provide any evidence to support an association between alpha_<1a>-adrenoceptor gene polymorphism and fern ale ure-thral syndrome, however, much larger studies of both Japan and U.S.are needed. Less
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