PROACROSIN ACROSIN DURING GUINEA-PIG SPERMATOGENESIS
PROACROSIN ACROSIN DURING GUINEA-PIG SPERMATOGENESIS
复制标题
DOI：
10.1016/0012-1606(88)90075-9
复制
发表时间：
1988-01-01
影响因子：
2.7
通讯作者：
GERTON, GL
中科院分区：
文献类型：
--
作者：
ARBOLEDA, CE;GERTON, GL
通过单位重力沉降速度分离富集的豚鼠生精细胞群。通过酶谱、免疫印迹和免疫荧光分析每个细胞群中顶体素原/顶体素家族成员的存在。在含有0.1%明胶的十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后，在圆形精子细胞提取物中检测到分子量为55,000（主要）和50,000（次要）的蛋白酶活性。浓缩精细胞提取物含有蛋白酶活性，分子量在 55,000 至 50,000 之间。这些主要蛋白酶活性的分子量与用针对纯化的公猪顶体蛋白的单特异性兔抗血清进行免疫印迹检测到的抗原相似。豚鼠精子提取物和离子载体 A23187 诱导顶体反应后释放的可溶性顶体成分含有三种主要蛋白酶活性（Mr 32,000、34,000、47,000），但只有 47,000 Mr 蛋白酶与抗体发生交叉反应。精子细胞和精子蛋白酶活性被来自其他物种的顶体蛋白活性的经典效应子抑制和激活。免疫荧光证明顶体素原/顶体素早在精子发生的高尔基期就存在。此外，免疫反应性以每个精子细胞阶段的特征方式局限于顶体。这些结果表明顶体素原/顶体素可以在最早的精子形成阶段通过电泳和免疫学技术检测到，并表明顶体素原/顶体素的分子量随着精子细胞的成熟而发生变化。
Enriched populations of guinea pig spermatogenic cells were isolated by sedimentation velocity at unit gravity. Each cell population was analyzed for the presence of members of the proacrosin/acrosin family by enzymography, immunoblotting, and immunofluorescence. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis in gels containing 0.1% gelatin, protease activities with molecular weights of 55,000 (major) and 50,000 (minor) were detected in round spermatid extracts. Condensing spermatid extracts contained protease activities with molecular weights between 55,000 and 50,000. These major protease activities had molecular weights similar to antigens detected by immunoblotting with a monospecific rabbit antiserum directed against purified boar acrosin. Extracts of guinea pig sperm and the soluble acrosomal components released following the acrosome reaction induced with ionophore A23187 contained three major protease activities (Mr 32,000, 34,000, 47,000) but only the 47,000 Mr protease cross-reacted with the antibody. The spermatid and sperm protease activities were inhibited and activated by classical effectors of acrosin activity from other species. Immunofluorescence demonstrated that proacrosin/acrosin was present as early as the Golgi phase of spermiogenesis. In addition, immunoreactivity was confined to the acrosomes in a manner characteristic of each spermatid stage. These results demonstrate that proacrosin/acrosin can be detected in the earliest spermiogenic stages by electrophoretic and immunological techniques and suggest that changes in the molecular weights of proacrosin/acrosin occur as spermatids mature.