Expression and Localisation of Aquaporin Water Channels in Human Urothelium In Situ and In Vitro
Expression and Localisation of Aquaporin Water Channels in Human Urothelium In Situ and In Vitro
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DOI：
10.1016/j.eururo.2008.08.013
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发表时间：
2009-12-01
期刊：
影响因子：
23.4
通讯作者：
Southgate, Jennifer
中科院分区：
文献类型：
--
作者：
Rubenwolf, Peter C.;Georgopoulos, Nikolaos T.;Southgate, Jennifer
背景：尿路上皮通常被认为对水和尿液成分是不渗透的。人的尿路上皮细胞表达水通道蛋白(AQP)水通道作为水和溶质运输的基础的可能性以前还没有被研究过。目的：研究AQP水通道在人原位尿路上皮细胞、增殖的尿路上皮细胞培养和分化的组织结构中的表达。设计、背景和对象：用逆转录聚合酶链式反应(RT-PCR)和免疫标记法检测人尿上皮细胞原位和培养的水通道蛋白的表达。从多个外科(膀胱和输尿管)标本中分离的新鲜尿路标本和培养中增殖和分化的正常人尿路上皮(NHU)细胞进行了表达筛选。结果和局限性：AQP3、AQP4、AQP7、AQP9和AQP11的转录本在新鲜分离的尿路和培养的NHU细胞中表达一致。AQP0、AQP1、AQP2、AQP5、AQP6、AQP8、AQP10和AQP12均不表达。免疫组织化学证实AQP3、AQP4、AQP7和AQP9在蛋白水平表达。AQP3在尿路上皮和体外分化的组织结构中的基底层和中间层均有较强的表达。结论：本研究首次证实AQP3在人尿路上皮细胞中表达，提示AQP3在尿路上皮水和溶质转运中可能起重要作用。我们的发现挑战了尿路作为非渗透转运和储存单元的传统概念，并为进一步研究水通道蛋白在人体尿路上皮中的生物学和临床相关性提供了一个通用的平台。(C)2008年欧洲泌尿外科协会。爱思唯尔出版公司版权所有。
Background: Urothelium is generally considered to be impermeable to water and constituents of urine. The possibility that human urothelium expresses aquaporin (AQP) water channels as the basis for water and solute transport has not previously been investigated.Objective: To investigate the expression of AQP water channels by human urothelium in situ, in proliferating urothelial cell cultures and in differentiated tissue constructs.Design, setting, and participants: AQP expression by human urothelium in situ and cultured urothelial cells was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunolabelling. Expression screening was carried out on samples of freshly isolated urothelia from multiple surgical (bladder and ureteric) specimens and on proliferating and differentiated normal human urothelial (NHU) cells in culture. Urothelial tissue constructs were established and investigated for expression of urothelial differentiation markers and AQPs.Measurements: Qualitative study.Results and limitations: Transcripts for AQP3, AQP4, AQP7, AQP9, and AQP11 were expressed consistently by freshly isolated urothelia as well as by cultured NHU cells. AQP0, AQP1, AQP2, AQP5, AQP6, AQP8, AQP10, and AQP12 were not expressed. Immunochemistry confirmed expression of AQP3, AQP4, AQP7, and AQP9 at the protein level. AQP3 was shown to be intensely expressed at cell borders in the basal and intermediate layers in both urothelium in situ and differentiated tissue constructs in vitro.Conclusions: This is the first study to demonstrate that AQPs are expressed by human urothelium, suggesting a potential role in transurothelial water and solute transport. Our findings challenge the traditional concept of the urinary tract as an impermeable transit and storage unit and provide a versatile platform for further investigations into the biological and clinical relevance of AQPs in human urothelium. (C) 2008 European Association of Urology. Published by Elsevier B. V. All rights reserved.