Functional characterization of transient receptor potential channels in mouse urothelial cells
Functional characterization of transient receptor potential channels in mouse urothelial cells
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DOI：
10.1152/ajprenal.00599.2009
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发表时间：
2010-03-01
影响因子：
4.2
通讯作者：
Nilius, Bernd
中科院分区：
文献类型：
--
作者：
Everaerts, Wouter;Vriens, Joris;Nilius, Bernd
Everaerts W，Vriens J，Owsianik G，Pastudino G，Voets T，De Ridder D，Nilius B.小鼠尿路上皮细胞瞬时受体电位通道的功能特性。美国肾脏生理学杂志298：F692-F701，2010年。首次发表于2009年12月16日; doi：10.1152/ajprenal.00599.2009。目前认为膀胱尿路上皮是一种感觉结构，有助于膀胱的机械感觉和化学感觉。瞬时受体电位（TRP）阳离子通道作为多模态传感器，可能是尿路上皮细胞某些接受特性的基础。然而，尿路上皮细胞的确切TRP通道表达谱尚不清楚。在这项研究中，我们进行了系统的分析，在小鼠尿路上皮细胞的各种TRP通道的分子和功能的表达。分离来自对照和trpv 4（-/-）小鼠的尿路上皮细胞，培养（12-48 h），并用于定量实时PCR、免疫细胞化学、钙成像和全细胞膜片钳实验。在mRNA水平上，TRPV 4、TRPV 2和TRPM 7是表达最丰富的TRP基因。免疫组化显示TRPV 4在质膜中有明显表达，而TRPV 2在细胞质中更突出。TRPM 7在质膜以及细胞质囊泡中被检测到。使用TRP通道激动剂和拮抗剂的钙成像和膜片钳实验提供了TRPV 4、TRPV 2和TRPM 7功能表达的证据，但TRPA 1、TRPV 1和TRPM 8没有。总之，我们已经证明了TRPV 4、TRPV 2和TRPM 7在小鼠尿路上皮细胞中的功能性表达。这些通道可能有助于尿路上皮层的（机械）感觉功能，并代表治疗膀胱功能障碍的潜在靶点。
Everaerts W, Vriens J, Owsianik G, Appendino G, Voets T, De Ridder D, Nilius B. Functional characterization of transient receptor potential channels in mouse urothelial cells. Am J Physiol Renal Physiol 298: F692-F701, 2010. First published December 16, 2009; doi:10.1152/ajprenal.00599.2009.-The bladder urothelium is currently believed to be a sensory structure, contributing to mechano- and chemosensation in the bladder. Transient receptor potential (TRP) cation channels act as polymodal sensors and may underlie some of the receptive properties of urothelial cells. However, the exact TRP channel expression profile of urothelial cells is unclear. In this study, we have performed a systematic analysis of the molecular and functional expression of various TRP channels in mouse urothelium. Urothelial cells from control and trpv4(-/-) mice were isolated, cultured (12-48 h), and used for quantitative real-time PCR, immunocytochemistry, calcium imaging, and whole cell patch-clamp experiments. At the mRNA level, TRPV4, TRPV2, and TRPM7 were the most abundantly expressed TRP genes. Immunohistochemistry showed a clear expression of TRPV4 in the plasma membrane, whereas TRPV2 was more prominent in the cytoplasm. TRPM7 was detected in the plasma membrane as well as cytoplasmic vesicles. Calcium imaging and patch-clamp experiments using TRP channel agonists and antagonists provided evidence for the functional expression of TRPV4, TRPV2, and TRPM7 but not of TRPA1, TRPV1, and TRPM8. In conclusion, we have demonstrated functional expression of TRPV4, TRPV2, and TRPM7 in mouse urothelial cells. These channels may contribute to the (mechano) sensory function of the urothelial layer and represent potential targets for the treatment of bladder dysfunction.