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水通道蛋白-2是一种受调控的水通道,表达于大鼠远端结肠上皮细胞的顶端膜中。

Aquaporin-2, a regulated water channel, is expressed in apical membranes of rat distal colon epithelium.

作者信息

Gallardo P, Cid L P, Vio C P, Sepúlveda F V

机构信息

Centro de Estudios Científicos, Valdivia, Chile.

出版信息

Am J Physiol Gastrointest Liver Physiol. 2001 Sep;281(3):G856-63. doi: 10.1152/ajpgi.2001.281.3.G856.

Abstract

Aquaporin-2 (AQP-2) is the vasopressin-regulated water channel expressed in the apical membrane of principal cells in the collecting duct and is involved in the urinary concentrating mechanism. In the rat distal colon, vasopressin stimulates water absorption through an unknown mechanism. With the hypothesis that AQP-2 could contribute to this vasopressin effect, we studied its presence in rat colonic epithelium. We used RT-PCR, in situ hybridization, immunoblotting, and immunocytochemistry to probe for AQP-2 expression. An AQP-2 amplicon was obtained through RT-PCR of colon epithelium RNA, and in situ hybridization revealed AQP-2 mRNA in colonic crypts and, to a lesser extent, in surface absorptive epithelial cells. AQP-2 protein was localized to the apical membrane of surface absorptive epithelial cells, where it colocalized with H(+)-K(+)-ATPase but not with Na(+)-K(+)-ATPase. AQP-2 was absent from the small intestine, stomach, and liver. Water deprivation increased the hybridization signal and the protein level (assessed by Western blot analysis) for AQP-2 in distal colon. This was accompanied by increased p-chloromercuriphenylsulfonic acid-sensitive water absorption. These results indicate that AQP-2 is present in the rat distal colon, where it might be involved in a water-sparing mechanism. In addition, these results support the idea that AQP-2, and probably other aquaporins, are involved in water absorption in the colon.

摘要

水通道蛋白-2(AQP-2)是一种受抗利尿激素调节的水通道,表达于集合管主细胞的顶端膜,参与尿液浓缩机制。在大鼠远端结肠,抗利尿激素通过未知机制刺激水的吸收。基于AQP-2可能参与抗利尿激素这一作用的假设,我们研究了其在大鼠结肠上皮中的存在情况。我们运用逆转录聚合酶链反应(RT-PCR)、原位杂交、免疫印迹和免疫细胞化学技术来检测AQP-2的表达。通过对结肠上皮RNA进行RT-PCR获得了AQP-2扩增子,原位杂交显示在结肠隐窝以及程度较轻的表面吸收上皮细胞中有AQP-2信使核糖核酸(mRNA)。AQP-2蛋白定位于表面吸收上皮细胞的顶端膜,在该处它与氢钾三磷酸腺苷酶(H(+)-K(+)-ATPase)共定位,但不与钠钾三磷酸腺苷酶(Na(+)-K(+)-ATPase)共定位。小肠、胃和肝脏中不存在AQP-2。禁水增加了远端结肠中AQP-2的杂交信号和蛋白水平(通过蛋白质免疫印迹分析评估)。这伴随着对氯汞苯磺酸敏感的水吸收增加。这些结果表明AQP-2存在于大鼠远端结肠,可能参与节水机制。此外,这些结果支持了AQP-2以及可能其他水通道蛋白参与结肠水吸收的观点。

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