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Tissue distribution of UT-A and UT-B mRNA and protein in rat.

机译:大鼠中UT-A和UT-B mRNA和蛋白质的组织分布。

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摘要

Mammalian urea transporters are facilitated membrane transport proteins belonging to two families, UT-A and UT-B. They are best known for their role of maintaining the renal inner medullary urinary concentrating gradient. Urea transporters have also been identified in tissues not typically associated with urea metabolism. The purpose of this study was to survey the major organs in rat to determine the distribution of UT-A and UT-B mRNA transcripts and protein forms and determine their cellular localization. Five kidney subregions and 17 extrarenal tissues were screened by Northern blot analysis using two UT-A and three UT-B probes and by Western blot analysis using polyclonal COOH-terminal UT-A and UT-B antibodies. Immunohistochemistry was performed on 16 extrarenal tissues using the same antibodies. In kidney, we detected mRNA transcripts and protein bands consistent with previously-identified UT-A and UT-B isoforms, as well as novel forms. We found that UT-A mRNA and protein are widely expressed in extrarenal tissues in various forms that are different from the known isoforms. We determined the cellular localization of UT-A and UT-B in these tissues. We found that both UT-A and UT-B are ubiquitously expressed as numerous tissue-specific mRNA transcripts and protein forms that are localized to cell membranes, cytoplasm, or nuclei.
机译:哺乳动物尿素转运蛋白是属于两个家族UT-A和UT-B的促进膜转运蛋白。他们以维持肾脏内髓质尿液浓度梯度的作用而闻名。还已经在通常不与尿素代谢相关的组织中鉴定出尿素转运蛋白。这项研究的目的是调查大鼠的主要器官,以确定UT-A和UT-B mRNA转录本的分布和蛋白质形式,并确定它们的细胞定位。通过使用两个UT-A和三个UT-B探针的Northern印迹分析和使用多克隆COOH末端UT-A和UT-B抗体的Western印迹分析,筛选了五个肾脏子区域和17个肾外组织。使用相同的抗体对16个肾外组织进行免疫组织化学。在肾脏中,我们检测到与先前确定的UT-A和UT-B同工型以及新颖形式一致的mRNA转录本和蛋白带。我们发现UT-A mRNA和蛋白在肾外组织中以不同于已知同工型的各种形式广泛表达。我们确定了UT-A和UT-B在这些组织中的细胞定位。我们发现UT-A和UT-B都以大量组织特异性mRNA转录本和蛋白质形式广泛表达,这些转录本和蛋白质形式定位于细胞膜,细胞质或细胞核。

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