Assessing the Role of Claudins in Maintaining the Integrity of Epididymal Tight Junctions Using Novel Human Epididymal Cell Lines
Assessing the Role of Claudins in Maintaining the Integrity of Epididymal Tight Junctions Using Novel Human Epididymal Cell Lines
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DOI：
10.1095/biolreprod.109.083196
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发表时间：
2010-06-01
影响因子：
3.6
通讯作者：
Cyr, Daniel G.
中科院分区：
文献类型：
--
作者：
Dube, Evemie;Dufresne, Julie;Cyr, Daniel G.
附睾负责睾丸后精子的成熟。精子的成熟依赖于附睾沿着的管腔微环境。虽然附睾的作用已经得到了很好的证实，但由于生物学工具有限，精子成熟的分子和细胞机制仍有待阐明，特别是在人类中。我们已经建立了第一个稳定的上皮细胞系转化SV 40大T抗原（LTAg）从两个地区的人成人附睾。细胞系由二倍体主细胞的同质群体组成，其具有与体内人主细胞相似的超微结构特征。这些细胞表达粘附蛋白（钙粘蛋白CDH 1和CDH 2）和紧密连接（紧密连接蛋白CLDN 1、CLDN 2、CLDN 3、CLDN 4、CLDN 7和CLDN 8）以及桥粒（桥斑蛋白，DSP）的转录物。在可生育的人附睾头细胞系1（FHCE 1）中的跨上皮电阻（TER）测量以及紧密连接蛋白1（TJP 1）、闭合蛋白和CLDN 1的免疫定位表明，这些细胞形成功能性紧密连接。此外，使用特异性siRNA敲低CLDN 1、CLDN 3、CLDN 4或CLDN 7导致TER显著降低，表明这些CLDN对于血液-附睾屏障的屏障功能是必需的。因此，CLDN 1、CLDN 3、CLDN 4和CLDN 7的破坏可能导致附睾功能障碍，导致男性不育。
The epididymis is responsible for posttesticular sperm maturation. Sperm maturation is dependent on the luminal microenvironments along the epididymis. Though the role of the epididymis is well established, the molecular and cellular mechanisms responsible for sperm maturation remain to be elucidated, particularly in the human, as limited biological tools exist. We have established the first stable epithelial cell lines transformed with SV40 large T antigen (LTAg) from two regions of the human adult epididymis. The cell lines are composed of homogenous populations of diploid principal cells that possess ultrastructural characteristics similar to those of human principal cells in vivo. These cells express transcripts for adherens (cadherins CDH1 and CDH2) and tight (claudins CLDN1, CLDN2, CLDN3, CLDN4, CLDN7, and CLDN8) junctions as well as desmosomes (desmoplakin, DSP). Transepithelial resistance (TER) measurements in fertile human caput epididymal cell line 1 (FHCE1) as well as the immunolocalization of tight junctional protein 1 (TJP1), occludin, and CLDN1 indicate that these cells form functional tight junctions. Furthermore, knockdown of CLDN1, CLDN3, CLDN4, or CLDN7 using specific siRNAs resulted in significant decreases in TER, suggesting that these CLDNs are essential for the barrier function of the blood- epididymis barrier. Disruption of CLDN1, CLDN3, CLDN4, and CLDN7 could, therefore, lead to epididymal dysfunction, resulting in male infertility.