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来自附睾的分离原代上皮细胞的全细胞膜片钳记录

Whole-cell Patch-clamp Recordings of Isolated Primary Epithelial Cells from the Epididymis.

作者信息

Zhang Bao Li, Gao Da Yuan, Zhang Xiao Xu, Shi Shuo, Shum Winnie

机构信息

School of Life Science and Technology, ShanghaiTech University; Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences; University of Chinese Academy of Sciences.

School of Life Science and Technology, ShanghaiTech University; University of Chinese Academy of Sciences.

出版信息

J Vis Exp. 2017 Aug 3(126):55700. doi: 10.3791/55700.

Abstract

The epididymis is an essential organ for sperm maturation and reproductive health. The epididymal epithelium consists of intricately connected cell types that are distinct not only in molecular and morphological features but also in physiological properties. These differences reflect their diverse functions, which together establish the necessary microenvironment for the post-testicular sperm development in the epididymal lumen. The understanding of the biophysical properties of the epididymal epithelial cells is critical for revealing their functions in sperm and reproductive health, under both physiological and pathophysiological conditions. While their functional properties have yet to be fully elucidated, the epididymal epithelial cells can be studied using the patch-clamp technique, a tool for measuring the cellular events and the membrane properties of single cells. Here, we describe the methods of cell isolation and whole-cell patch-clamp recording to measure the electrical properties of primary dissociated epithelial cells from the rat cauda epididymides.

摘要

附睾是精子成熟和生殖健康的重要器官。附睾上皮由错综复杂连接的细胞类型组成,这些细胞类型不仅在分子和形态特征上不同,而且在生理特性上也不同。这些差异反映了它们多样的功能,共同为附睾管腔内睾丸后精子发育建立了必要的微环境。了解附睾上皮细胞的生物物理特性对于揭示其在生理和病理生理条件下对精子和生殖健康的作用至关重要。虽然它们的功能特性尚未完全阐明,但可以使用膜片钳技术研究附睾上皮细胞,该技术是一种用于测量单细胞的细胞事件和膜特性的工具。在这里,我们描述了细胞分离和全细胞膜片钳记录的方法,以测量来自大鼠附睾尾部的原代解离上皮细胞的电特性。

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Physiology of epithelial chloride and fluid secretion.上皮细胞氯离子和液体分泌的生理学。
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