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水通道蛋白-7在大鼠胃肠道中的表达及免疫定位

Expression and immunolocalization of aquaporin-7 in rat gastrointestinal tract.

作者信息

Laforenza Umberto, Gastaldi Giulia, Grazioli Monica, Cova Emanuela, Tritto Simona, Faelli Alide, Calamita Giuseppe, Ventura Ulderico

机构信息

Department of Experimental Medicine, Section of Human Physiology, University of Pavia, Via Forlanini 6, I-27100 Pavia, Italy.

出版信息

Biol Cell. 2005 Aug;97(8):605-13. doi: 10.1042/BC20040090.

Abstract

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.

RESULTS

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.

CONCLUSIONS

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.

摘要

背景信息

在哺乳动物的胃肠道中,水既可以随消化液分泌,也可以被小肠和大肠吸收。跨细胞的水转运可由水通道蛋白(AQPs)这一跨膜蛋白家族介导,最近在胃肠道中也已得到证实。然而,水通道蛋白在胃肠道中的定位、表达和功能尚未完全明确。在本研究中,我们调查了:(1)通过半定量逆转录聚合酶链反应和免疫印迹法检测大鼠胃肠道某些部位水通道蛋白7(AQP7)的表达;(2)通过免疫组织化学法检测AQP7的细胞和亚细胞定位。

结果

AQP7 mRNA和蛋白在小肠中高表达,在盲肠、结肠和直肠中弱表达,在胃中未表达。使用大鼠胃肠道膜组分进行的免疫印迹分析显示,AQP7蛋白有两条主要条带,分子量约为34 kDa和40 kDa。当抗AQP7抗体与免疫原性肽预先吸附时,未观察到条带。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强烈的AQP7标记,而在隐窝细胞中标记较弱或未观察到。标记尤其在顶端膜明显,但也观察到细胞内染色。

结论

结果表明AQP7存在于小肠和大肠中。AQP7蛋白在浅表上皮细胞顶端的高表达表明其参与了通过绒毛上皮的快速液体转运。

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