Flörke-Gerloff S, Töpfer-Petersen E, Müller-Esterl W, Schill W B, Engel W
Hum Genet. 1983;65(1):61-7. doi: 10.1007/BF00285030.
Using the indirect immunofluorescent staining technique, the developmental patterns of (pro) acrosin and the outer acrosomal membrane were studied in human spermatogenesis. Specific antibodies against purified acrosin and outer acrosomal membranes from boar spermatozoa were raised in the rabbit and were found to crossreact with (pro)acrosin and outer acrosomal membrane from human spermatogenic cells. It was concluded that (pro)acrosin as well as the molecules building up the outer acrosomal membrane have been highly conserved during mammalian evolution. In the course of human spermatogenesis (pro)acrosin as well as the outer acrosomal membrane first appear in the haploid spermatids; the fluorescent areas of the individual cells steadily increase during spermiogenesis. Staining for acrosin and the outer acrosomal membrane, respectively, was found in identical compartments of the spermatogenic cells in juxtaposition to the nucleus. Round-headed spermatozoa from an infertile patient did not stain for (pro)acrosin or outer acrosomal membrane. The lack of the acrosin system was further substantiated by the gelatin substrate film technique demonstrating the absence of a gelatinolytic protease in round-headed spermatozoa. Hence, round-headed spermatozoa lack the acrosome with its constituent membrane proteins and the acrosin system housed by the acrosome of normal spermatozoa.
运用间接免疫荧光染色技术,对人类精子发生过程中(原)顶体蛋白酶和顶体外膜的发育模式进行了研究。用来自公猪精子的纯化顶体蛋白酶和顶体外膜制备的特异性抗体免疫家兔,发现这些抗体可与人精子发生细胞中的(原)顶体蛋白酶和顶体外膜发生交叉反应。由此得出结论,在哺乳动物进化过程中,(原)顶体蛋白酶以及构成顶体外膜的分子高度保守。在人类精子发生过程中,(原)顶体蛋白酶和顶体外膜首先出现在单倍体精子细胞中;在精子形成过程中,单个细胞的荧光区域稳步增加。在紧邻细胞核的精子发生细胞的相同区域分别检测到顶体蛋白酶和顶体外膜的染色。一名不育患者的圆头精子未检测到(原)顶体蛋白酶或顶体外膜的染色。明胶底物膜技术进一步证实了顶体蛋白酶系统的缺失,该技术显示圆头精子中不存在明胶分解蛋白酶。因此,圆头精子缺乏正常精子顶体所含的顶体及其组成膜蛋白,也缺乏顶体蛋白酶系统。