Newton S C, Millette C F
Department of Cell Biology and Neurosciences, University of South Carolina, Columbia 29208.
J Androl. 1992 Mar-Apr;13(2):160-71.
This study concerns Sertoli cell-spermatogenic cell adhesive interactions in the seminiferous tubule. Sertoli cell surface polypeptides involved in germ cell-Sertoli cell adhesion were identified by serological inhibition of an in vitro Sertoli-germ cell adhesion assay. This assay was modified from a previously reported adhesion assay, and employs a scanning laser cytometer for quantification of adherent cells. Reactivity of the polyclonal antiserum raised against rat Sertoli cells was also assessed via immunofluorescent microscopy. The addition of antiserum to the adhesion assay resulted in a 42% to 66% inhibition of cell-cell adhesion. Moreover, preincubation of antiserum with Sertoli cell monolayers resulted in a significant reduction of spermatogenic cell binding. Conversely, preincubation of antiserum with germ cells resulted in no reduction. Western blot analysis of the antiserum against purified Sertoli cell membranes indicated reactivity with four polypeptides. The data suggest that one or more of these polypeptides are directly involved in the adhesion of germ cells to Sertoli cell monolayers in vitro.
本研究关注生精小管中支持细胞与生精细胞的黏附相互作用。通过体外支持细胞 - 生殖细胞黏附试验的血清学抑制,鉴定了参与生殖细胞与支持细胞黏附的支持细胞表面多肽。该试验是在先前报道的黏附试验基础上改进的,采用扫描激光细胞仪对黏附细胞进行定量分析。还通过免疫荧光显微镜评估了针对大鼠支持细胞产生的多克隆抗血清的反应性。在黏附试验中加入抗血清导致细胞间黏附受到42%至66%的抑制。此外,抗血清与支持细胞单层预孵育导致生精细胞结合显著减少。相反,抗血清与生殖细胞预孵育则未导致减少。针对纯化的支持细胞膜的抗血清的蛋白质印迹分析表明与四种多肽有反应性。数据表明这些多肽中的一种或多种在体外直接参与生殖细胞与支持细胞单层的黏附。