【作者】 秦昆；

【导师】 康现江；

【作者基本信息】 河北大学 ， 生物化学与分子生物学， 2007， 硕士

【摘要】 动物精子形成过程中,精子细胞形成精子,形态结构发生了显著变化。精子由圆形变为长形,染色质高度浓缩,这个过程中核碱性蛋白对浓缩精子核和保护DNA遗传信息起了重要作用。细胞核中与DNA结合的碱性蛋白也发生了替代或更替,这对遗传物质的稳定性和精子完成受精过程至关重要。因此细胞核中的碱性蛋白对于精子形成、受精过程等生殖功能有重要的作用。不同物种之间核碱性蛋白种类和数量有所不同。氨基酸组成和各个氨基酸所占比例差异性也很大。这与物种本身所处进化地位有密切关系。尤其是甲壳动物精子,该类精子是非浓缩核,呈现几乎裸露的状态。这一现象在十足目中是普遍存在的,由此吸引了众多研究学者。中国明对虾(Fenneropenaeus chinensis)精子形成过程中核碱性蛋白种类、核碱性蛋白替代特征的报道较少。本文通过SDS聚丙烯酰胺凝胶电泳和酸性尿素聚丙烯酰胺凝胶电泳,对其核碱性蛋白的生化性质进行了初步分析,对十足目动物精子形成过程碱性蛋白的替代特征进行了探讨。结果如下：肌细胞与生精细胞的核碱性蛋白都有5种,H1、H2A、H2B、H3、H4；H1有2条带。精子核中没有观察到碱性蛋白。根据核碱性蛋白电泳迁移率判断其分子量由小到大为H4、H2A、H2B、H3、H1。而在AUT-PAGE中,由于加入非离子型去污剂Triton X-100,其与碱性蛋白形成复合物,使蛋白的迁移率产生明显变化,迁移率由大到小为H4、H2B、H3、H1、H2A。生精细胞时期,碱性蛋白主要集中在细胞核的染色体上；精子核中却没有发现碱性蛋白的存在。结合电镜方面的研究推测：中国明对虾精子形成过程中,碱性蛋白由细胞核转移到顶体囊中。更多还原

【Abstract】 During spermiogenesis in animals, spermatids change into spermatozoa. Their morphosis changes greatly. Nuclear basic proteins that are responsible for chromatin condensation and conservative for hereditary information of DNA are very vital in the process. Round spermatid will be elongated, and its chromatin undergoes substantial compaction during spermiogenesis. There is also a replacement of basic proteins which associate with DNA in the nucleus. The basic proteins’replacement is very important in maintaining the stability of genetic material and performing fertilization of sperm. Therefore, nuclear basic proteins play a critical role in the spermiogenesis, fertilization, and other reproduction functions.These findings displayed all sorts of amino acid composition, quantity and proportion, which is important for species evolution. Especially, Decapoda spermatozoa are different from that of most animals, with nucleus being non-condensing. The phenomenon is wide-spread in Decapods, and attracts many scientists to research. The results were those:There are five kinds of nuclear basic protein (H1,H2A,H2B,H3 and H4, H1 shown 2 bands), in muscle tissue and spermatogenic cells. Basic protein was invisible in sperm nucleus. Ruled by electrophoretic mobility of nuclear basic protein, the molecular weight of them arranged from large to small:H4, H2B, H2A, H3, H1. Under condition of AUT-PAGE, above sequence of nuclear basic proteins chaged into H4、H2B、H3、H1、H2A. There was a possible reason that the non-ionic detergent Triton X-100, which was poured, would form compounds with histones.Fenneropenaeus chinensis is economic crab in China, with the common characterized of Decapods. It is unknown about category and character of nuclear basic proteins displacement in spermiogenesis. The biochemistry character of the histones of F. chinensis was also analyzed preliminarily, to research the replacement character of the basic proteins during spermiogenesis in Decapoda, by the methods of SDS-PAGE and AU-PAGE.In the stage of spermatogenic cell, the basic proteind were kepted in the nuclear chromo-some. However, the basic proteins were not found in the sperm nucleus of F. chinensis. It could be presumed that the basic proteins migrated to the acrosomal vesicle from the nucleus during sperm formation in F. chinensis, with the research by electron microscope.更多还原