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- W2188713969 abstract "In recent years, both molecular biological and immunohistochemical techniques, utilizing receptor subtype-specific probes and antibodies to cloned central nervous system dopamine receptors, have revealed their presence in a number of peripheral organs and tissues. Molecular techniques have been hindered by the low abundance of receptor mRNA in these sites, and reverse transcription-polymerase chain reaction (RT-PCR) has been utilized to address this problem. However, RT-PCR is most often employed on either isolated mRNA or microdissected tissue samples, thereby limiting interpretation of whole tissue distribution. The present paper describes the use of a novel self-sustained sequence replication system (3SR) to amplify a target mRNA sequence in situ within the tissue or cell of interest that is then detected with the use of an internal labeled probe, using standard nonisotopic in situ hybridization. Specifically, D 1A receptor mRNA was amplified and detected in kidney sections of Wistar-Kyoto rats (WKY). The amplified D 1A receptor mRNA was localized to renal arterioles, juxtaglomerular apparatus, and both proximal and distal tubules. mRNA was colocalized to regions shown also to contain D 1A receptor protein. D 1A receptor mRNA was predominantly localized in the cortex. Specificity of D 1A receptor mRNA detection was confirmed by appropriate localization in rat brain sections known to express D 1A receptor mRNA. In addition, we confirmed the presence of renal D 1A receptor mRNA by RT-PCR. We conclude that D 1A receptor mRNA is expressed in a site-specific manner in the WKY kidney. The use of 3SR in situ permits elucidation of site specific mRNA localization in a manner not reported previously." @default.
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- W2188713969 date "1998-01-01" @default.
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- W2188713969 title "Detection of dopamine receptor D1Asubtype-specific mRNA in rat kidney by in situ amplification" @default.
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- W2188713969 doi "https://doi.org/10.1152/ajprenal.1998.274.1.f232" @default.
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