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Expression and immunolocalization of aquaporin-7 in rat gastrointestinal tract

机译:水通道蛋白7在大鼠胃肠道中的表达及免疫定位

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摘要

Background information. In the gastrointestinal tract of mammals, water can either be secreted with digestive juices or absorbed by the small and large intestine. Transcellular water movement can be mediated by the transmembrane protein family of AQPs (aquaporins), as has also been recently identified in the gastrointestinal tract. However, the localization, expression and functioning of AQPs in the gastrointestinal tract have not been completely characterized. For the present study, we investigated: (1) the expression of AQP7 in some portions of rat gastrointestinal tract by semiquantitative reverse transcriptase-PCR and by immunoblotting and (2) the cellular and subcellular localization of AQP7 by immunohistochemistry. \ud\udResults. AQP7 mRNA and proteins were highly expressed in the small intestine, weakly in the caecum, colon and rectum and were absent in the stomach. Immunoblotting analysis using rat gastrointestinal tract membrane fractions showed two major bands corresponding to a molecular mass of approx. 34 and 40 kDa for the AQP7 protein. No bands were observed when the anti-AQP7 antibody was preadsorbed with the immunizing peptide. Immunohistochemistry revealed strong AQP7 labelling in the surface epithelial cells of duodenum, jejunum, ileum, caecum, colon and rectum, whereas weak or no labelling was observed in the crypt cells. The labelling was manifest particularly in the apical membrane but intracellular staining was also observed. \ud\udConclusions. The results indicate that AQP7 is present in the small and large intestine. The higher expression of AQP7 protein at the apical pole of the superficial epithelial cells suggests its involvement in rapid fluid movement through the villus epithelium.
机译:背景信息。在哺乳动物的胃肠道中,水可以通过消化液分泌,也可以被大小肠吸收。近来在胃肠道中也发现,跨细胞的水运动可以由AQPs(水通道蛋白)的跨膜蛋白家族介导。但是,AQPs在胃肠道中的定位,表达和功能尚未完全表征。对于本研究,我们调查:(1)通过半定量逆转录酶-PCR和免疫印迹法在大鼠胃肠道的某些部位表达AQP7,以及(2)通过免疫组织化学对AQP7进行细胞和亚细胞定位。 \ ud \ ud结果。 AQP7 mRNA和蛋白在小肠中高表达,在盲肠,结肠和直肠中微弱表达,在胃中不存在。使用大鼠胃肠道膜级分的免疫印迹分析显示了两个主要谱带,其对应的分子量约为。 AQP7蛋白分别为34和40 kDa。当抗AQP7抗体被免疫肽预吸附时,没有观察到条带。免疫组织化学显示十二指肠,空肠,回肠,盲肠,结肠和直肠的表面上皮细胞中有较强的AQP7标记,而在隐窝细胞中则观察到弱或没有标记。标记尤其在顶膜中明显,但也观察到细胞内染色。 \ ud \ ud结论。结果表明,AQP7存在于小肠和大肠中。 AQP7蛋白在浅层上皮细胞的顶极高表达,表明其参与通过绒毛上皮的快速液体运动。

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