Klinefelter G R, Roberts N L, Suarez J D
United States Environmental Protection Agency Health Effects Research Laboratory, Research Triangle Park, North Carolina.
J Androl. 1992 Sep-Oct;13(5):409-21.
It was recently demonstrated that the Leydig cell toxicant ethane dimethanesulphonate (EDS) produces multiple effects on the epididymis after a single in vivo exposure. To determine whether any of the perturbations were mediated by a direct action of the compound, we used a novel system for the coculture of epididymal epithelial cells and sperm from the caput epididymidis. This system maintains the morphologic integrity and cell polarity of the epididymal epithelial cells before and during coculture, and the sperm recovered after coculture have intact plasma and acrosomal membranes. In addition, several functions required for epididymal sperm maturation are expressed, including the secretion of protein by the epididymal epithelium, the association of secreted protein with the plasma membrane of cocultured sperm, and the acquisition of progressive motility by cocultured sperm. In vitro exposure of epididymal epithelial cells and sperm to EDS results in a significant decline in protein secretion by the epithelial cells during coculture, and in particular, a dose-dependent decline in a 36- to 38-kd protein (PI 4.0 to 4.5) and a 34- to 36-kd protein (PI 4.5 to 5.0). Moreover, these and other proteins are not recovered from the sperm membrane of cocultured sperm after EDS treatment. Finally, EDS results in a dose-dependent decline in the percentage of both motile and progressively motile sperm recovered after coculture compared with that of sperm from untreated cocultures. These effects on sperm motility were not observed when sperm were pretreated with EDS and subsequently cocultured with untreated epithelial cells. We conclude that EDS alters epididymal sperm maturation by acting directly on the epididymal epithelium to mediate changes in sperm membrane protein, and that this may subsequently alter the development of the progressive motility of sperm.
最近有研究表明，莱迪希细胞毒性剂乙烷二甲磺酸盐（EDS）在体内单次暴露后会对附睾产生多种影响。为了确定这些扰动是否由该化合物的直接作用介导，我们使用了一种新的系统来共培养附睾上皮细胞和来自附睾头的精子。该系统在共培养之前和期间维持附睾上皮细胞的形态完整性和细胞极性，并且共培养后回收的精子具有完整的质膜和顶体膜。此外，还表达了附睾精子成熟所需的几种功能，包括附睾上皮分泌蛋白质、分泌的蛋白质与共培养精子的质膜结合，以及共培养精子获得渐进性运动能力。附睾上皮细胞和精子在体外暴露于EDS会导致共培养期间上皮细胞蛋白质分泌显著下降，特别是36至38kd蛋白质（PI 4.0至4.5）和34至36kd蛋白质（PI 4.5至5.0）呈剂量依赖性下降。此外，EDS处理后，这些蛋白质和其他蛋白质未从共培养精子的精子膜中回收。最后，与未处理的共培养精子相比，EDS导致共培养后回收的活动精子和渐进性活动精子的百分比呈剂量依赖性下降。当精子先用EDS预处理，然后与未处理的上皮细胞共培养时，未观察到对精子运动的这些影响。我们得出结论，EDS通过直接作用于附睾上皮来介导精子膜蛋白的变化，从而改变附睾精子成熟，这可能随后改变精子渐进性运动的发育。
