Martínez-Menárguez J A, Geuze H J, Ballesta J
Department of Cell Biology, Center for Electronmicroscopy, Utrecht University, The Netherlands.
J Histochem Cytochem. 1996 Apr;44(4):313-20. doi: 10.1177/44.4.8601690.
We studied the biogenesis of the acrosome in sperm cells in immunogold-labeled ultrathin cryosections of rat testis, using a variety of antibodies against endosomal/lysosomal marker protein and acrosin, the major secretory protein of sperm cells. As expected, acrosomes and proacrosomal vesicles in the trans-Golgi region contained abundant acrosin. Rat lysosomal membrane glycoprotein (lgp) 120 and mouse lysosome-associated membrane protein-1 were not detectable in the acrosomal membrane. Similarly, the late endosomal markers cation-dependent and -independent mannose 6-phosphate receptors were absent from the acrosome and proacrosomal vesicles. Therefore, acrosomes do not exhibit these endosomal/lysosomal features. Apart from (pro) acrosomal vesicles, both spermatocytes and spermatids contained classical lysosomes (positive for rat lgp 120, mouse lysosome-associated membrane protein-1, and cathepsin D) that were negative for acrosin. Quantitative analysis of the immunogold labeling showed that spermatocytes express more mannose 6-phosphate receptors and lgp 120 than spermatids, whereas the opposite situation existed for acrosin. These data indicate differential synthetic activity of lysosomal and acrosomal constituents in different states of sperm differentiation. Together, our observations argue against a lysosomal /endosomal origin of the acrosome.
我们使用多种针对内体/溶酶体标记蛋白和顶体蛋白酶(精子细胞的主要分泌蛋白)的抗体,在大鼠睾丸免疫金标记超薄冰冻切片中研究了精子细胞顶体的生物发生。正如预期的那样,反式高尔基体区域的顶体和前顶体小泡含有丰富的顶体蛋白酶。在顶体膜中未检测到大鼠溶酶体膜糖蛋白(lgp)120和小鼠溶酶体相关膜蛋白-1。同样,晚期内体标记物阳离子依赖性和非依赖性甘露糖6-磷酸受体在顶体和前顶体小泡中也不存在。因此,顶体不具有这些内体/溶酶体特征。除了(前)顶体小泡外,精母细胞和精子细胞都含有经典的溶酶体(对大鼠lgp 120、小鼠溶酶体相关膜蛋白-1和组织蛋白酶D呈阳性),而对顶体蛋白酶呈阴性。免疫金标记的定量分析表明,精母细胞比精子细胞表达更多的甘露糖6-磷酸受体和lgp 120,而顶体蛋白酶的情况则相反。这些数据表明在精子分化的不同状态下,溶酶体和顶体成分的合成活性存在差异。总之,我们的观察结果反对顶体起源于溶酶体/内体的观点。