Study On The Physiological And Biochemical Characteristic Changes Post-insemination And During Acrosome Reaction In The Sperm Of Chinese Shrimp, Fenneropenaeus Chinensis

By using cytobiology and biochemistry methods, we studied the wall structure of the seminal receptacle and the proteins of egg water and ultrastructure and protein composition changes during acrosome reaction of the sperm in Chinese shrimp, Fenneropenaeus chinensis. A few of physiological and biochemical characteristics that can affect sperm capacitation and associate with acrosome reaction were found. These provide experimental basis for advanced research in the mechanism of sperm capacitation and acrosome reaction in Chinese shrimp, F. chinensis.Thelycum wall was studied by histotomy. Its structure is similar to the primary structure of epidermis of shrimp although great differentiation occurred. There is great difference of structure character in different position of thelycum. The wall and crunode are thick in the entrance but thin inside. A single antrum in thelycum, one spermatophore lie in either side, and the interspace filled in by jel.Capillary zone electrophoresis was used to detect the biochemical components of egg water. Egg water was composed of two different components, which were also demonstrated by SDS-PAGE. The molecular weights of the two kinds of egg jellies are both about 200kDa.Gelatin-substrate SDS-PAGE results did not show any hydrolytic enzyme activity in egg water.Egg water was used to induce acrosome reaction in mature sperm of Fenneropenaeus chinensis. Transmission electron microscope and SDS-PAGE were used to study the ultrastructure and protein changes of the sperm in F. chinensis, which has undergone acrosome reaction. The results demonstrated that the sperms from female thelycum could be induced acrosome reaction in vitro by egg water, which was a kind of egg jelly released from oocyte into seawater during oocyte activation. More than fifty percent of sperm finished acrosome reaction during the first 30 min, in vitro. The whole event consists of the retraction of the spike and acrosome exocytosis. Spike was retracted and fused to acrosome cap followed by the release of acrosome granules. After acrosome reaction, many sorts of proteins in sperm are degraded. When the reacted sperms were examined with gelatin-substrate SDS-PAGE, six major peptide bands with hydrolytic enzyme activity were detected. Their molecular weights are 200kDa, 130 kDa, 66 kDa, 53 kDa, 48 kDa and 41 kDa respectively. These hydrolases cannot be detected in the sperms before acrosome reaction.At the same time, primary study were made in the research of sperm proteomics in F. chinensis. Raise a mathod of twice dissolution with different pH Lysis in sperm sample preparation, and found 2-D electrophoresis technology in the research of sperm proteomics in F. chinensis.