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秀丽隐杆线虫水通道蛋白基因家族的功能分析

Functional analysis of the aquaporin gene family in Caenorhabditis elegans.

作者信息

Huang Chunyi George, Lamitina Todd, Agre Peter, Strange Kevin

机构信息

Department of Biological Chemistry, Johns Hopkins University, Baltimore, Maryland, USA.

出版信息

Am J Physiol Cell Physiol. 2007 May;292(5):C1867-73. doi: 10.1152/ajpcell.00514.2006. Epub 2007 Jan 17.

Abstract

Aquaporin channels facilitate the transport of water, glycerol, and other small solutes across cell membranes. The physiological roles of many aquaporins remain unclear. To better understand aquaporin function, we characterized the aquaporin gene family in the nematode Caenorhabditis elegans. Eight canonical aquaporin-encoding genes (aqp) are present in the worm genome. Expression of aqp-2, aqp-3, aqp-4, aqp-6, or aqp-7 in Xenopus oocytes increased water permeability five- to sevenfold. Glycerol permeability was increased three to sevenfold by expression of aqp-1, aqp-3, or aqp-7. Green fluorescent protein transcriptional and translational reporters demonstrated that aqp genes are expressed in numerous C. elegans cell types, including the intestine, excretory cell, and hypodermis, which play important roles in whole animal osmoregulation. To define the role of C. elegans aquaporins in osmotic homeostasis, we isolated deletion alleles for four aqp genes, aqp-2, aqp-3, aqp-4, and aqp-8, which are expressed in osmoregulatory tissues and mediate water transport. Single, double, triple, and quadruple aqp mutant animals exhibited normal survival, development, growth, fertility, and movement under normal and hypertonic culture conditions. aqp-2;aqp-3;aqp-4;aqp-8 quadruple mutants exhibited a slight defect in recovery from hypotonic stress but survived hypotonic stress as well as wild-type animals. These results suggest that C. elegans aquaporins are not essential for whole animal osmoregulation and/or that deletion of aquaporin genes activates mechanisms that compensate for loss of water channel function.

摘要

水通道蛋白通道促进水、甘油和其他小分子溶质跨细胞膜运输。许多水通道蛋白的生理作用仍不清楚。为了更好地理解水通道蛋白的功能,我们对线虫秀丽隐杆线虫中的水通道蛋白基因家族进行了表征。线虫基因组中存在8个典型的水通道蛋白编码基因(aqp)。在非洲爪蟾卵母细胞中表达aqp - 2、aqp - 3、aqp - 4、aqp - 6或aqp - 7可使水通透性提高5至7倍。通过表达aqp - 1、aqp - 3或aqp - 7,甘油通透性提高了3至7倍。绿色荧光蛋白转录和翻译报告基因表明,aqp基因在秀丽隐杆线虫的多种细胞类型中表达,包括在整个动物渗透压调节中起重要作用的肠道、排泄细胞和皮下组织。为了确定秀丽隐杆线虫水通道蛋白在渗透稳态中的作用,我们分离了四个在渗透压调节组织中表达并介导水运输的aqp基因aqp - 2、aqp - 3、aqp - 4和aqp - 8的缺失等位基因。在正常和高渗培养条件下,单、双、三、四重aqp突变动物表现出正常的存活、发育、生长、繁殖力和运动能力。aqp - 2;aqp - 3;aqp - 4;aqp - 8四重突变体在从低渗应激中恢复时表现出轻微缺陷,但与野生型动物一样能在低渗应激中存活。这些结果表明,秀丽隐杆线虫水通道蛋白对于整个动物的渗透压调节并非必不可少,和/或水通道蛋白基因的缺失激活了补偿水通道功能丧失的机制。

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