雌核发育银鲫和两性生殖彩鲫精子蛋白组份的比较研究

A COMPARATIVE OF SPERM PROTEINS BETWEEN MALES FROM GYNOGENETIC AND GONOCHORISTIC CRUCIAN CARP

对雌核发育银鲫和两性融合彩鲫精子蛋白组份进行了比较分析。通过分级抽提得到精子的不同组份精浆、精头的膜、鞭毛和脱膜精头等 ,然后经不同的凝胶电泳系统 ,比较分析了银鲫精子和其两性亲缘种彩鲫精子相应组份可溶性蛋白成份的差异。研究表明 ,经分级抽提的银鲫精子和彩鲫精子的各个组份都含有其特定的蛋白谱带。精浆蛋白在两种鱼之间和两种鱼的不同个体之间都存在一定差异。精头膜、鞭毛和脱膜精头的可溶性蛋白在同种鱼不同个体间高度一致 ,但在两种鱼之间表现出差异。两种鱼精头膜的可溶性蛋白在SDS PAGE电泳图谱上基本一致 ,而在非变性聚丙烯酰胺凝胶电泳图谱上则具有各自的特征性谱带。鞭毛可溶性蛋白的SDS PAGE分析在雌核发育银鲫中揭示出一条特异的蛋白带。脱膜精头的可溶性蛋白在SDS PAGE电泳图谱上差异明显 ,存在几条特征性蛋白带 ,并经Acid UreaPAGE系统分析 ,证实这些特征性蛋白为碱性蛋白。这些发现为进一步鉴定雌核发育银鲫雄鱼精子的特异性蛋白和揭示其分子机制打下了基础。

In order to reveal the difference of sperm proteins between males from gynogenetic Silver crucian carp ( Carassius auratus gibelio ) and gonochoristic transparent Color crucian carp ( Carassius auratus transparent colored varity), the two types of sperm were respectively extracted by the same methods at same conditions, and various fractions were collected. The fractions include sperm plasma, sperm head membrane, flagella, and demembranated sperm head. The soluble protein components of these extracted fractions and the whole sperm were analysed by means of different polyacrylamide gel electrophoresis system, such as nondenaturing polyacrylamide gel electrophoresis, SDS PAGE and Acid Urea PAGE system. The obtained results show that each fraction contains specific soluble protein components, and a few of soluble proteins are different in some fraction between gynogenetic Silver crucian carp and gonochoristic Color crucian carp. The protein components of sperm plasma are different not only between the two kinds of fishes, but also among the individuals in the same kind of fish. It is suggested that the difference of sperm plasma proteins could be resulted from the contamination of blood and body fluid. In the flagella fraction, one soluble protein component specific for gynogenetic Silver crucain carp was observed by SDS PAGE system. The soluble protein components in sperm head membrane fraction were analysed by SDS PAGE system and nondenaturing PAGE system, and found that they have same patterns in SDS PAGE system but various pattern in nondenaturing PAGE system. It was suggested that the difference might be related to complex and functional status of the head membrane proteins. The significant difference of the soluble protein components between two types of fishes were discovered in demembranated sperm head fraction. Several protein bands specific for gynogenetic Silver crucain carp were revealed by SDS PAGE analysis. Furthermore, the specific proteins were demonstrated to be basic proteins by Acid Urea PAGE system. These discoveries laid a foundation for identifying specific proteins and revealing the mloe cular mechanism in gynogenetic Silver crucian carp sperm