THE SPERM SURFACE IN SPERMATOGENESIS AND FERTILIZATION

这项研究计划的总体目标是确定 精子表面通过精子发生分化以及精子是如何 受精后，卵表面成分分散在卵子表面。 更具体的目标包括：(1)使用单抗 针对特定部位的精子表面抗原，以确定在 精子发生以及这些抗原在睾丸中被添加到 生精细胞表面；(2)克隆这些细胞的编码基因 位点特异性表面抗原，并使用这些克隆的基因，(3) 确定精子发生的时间和在睾丸中的位置 基因转录；(4)准备和筛选合适的表达 可从中分离这些基因的文库，(5)亚克隆 将基因导入适当的转录载体，以及(6)设计 有效的分段技术，实现现场最终确定 杂交和抗体结合研究；(7)确定 部位特异性精子表面抗原横向分散在精子表面 多精受精卵，如果是，(8)以确定 细胞骨架和代谢抑制剂对这一侧方的影响 分散；以及，使用最近开发的融合鸡蛋的程序(9) 为了确定精子表面抗原是否均匀地分散在未受精的- 受精和受精-受精卵融合。 由于将要研究的表面抗原具有钙离子通道 能力和已知参与精子顶体反应 和精子-卵子结合，这些目标和决定应该提供 更深入地了解精子发生、精子受精率和 不孕不育，精子-卵子结合，这些目标和决定应该 更深入地了解精子发生、精子受精率和 不育症，精卵结合，精子成分转移到 受精卵和早期精子成分的功能 胚胎。在更一般的生物学术语中，它们应该提供 关于质膜运动的其他基本信息 分化细胞中的蛋白质，之间的关系 细胞骨架与膜流动性、膜融合与细胞 去压抑。

The general objectives of this research plan are to determine how the sperm surface differentiates through spermatogenesis and how sperm surface components disperse in the egg surface after fertilization. More specific aims include: (1) the use of monoclonal antibodies against site-specific sperm surface antigens to determine when during spermatogenesis and where in the testis these antigens are added to the surfaces of spermatogenic cells; (2) to clone the genes coding for these site-specific surface antigens and, using these cloned genes, (3) to determine when during spermatogenesis and where in the testis these genes are transcribed; (4) to prepare and screen appropriate expression libraries from which these genes may be isolated, (5) to subclone these genes into appropriate transcription vectors, and (6) to devise effective sectioning techniques to carry out definitive in situ hybridization and antibody binding studies; (7) to determine whether site-specific sperm surface antigens disperse laterally on the surfaces of polyspermically fertilized eggs and, if so, (8) to determine the effects of cytoskeletal and metabolic inhibitors on this lateral dispersal; and, using recently developed procedures for fusing eggs, (9) to determine if sperm surface antigens disperse evenly in unfertilized- fertilized and fertilized-fertilized egg fusions. Since the surface antigens that will be studied possess Ca2+ -channeling capabilities and are known to be involved in the sperm acrosome reaction and sperm-egg binding, these aims and determinations should provide closer understandings of spermatogenesis, sperm fertility and infertility, sperm-egg binding, these aims and determinations should provide closer understandings of spermatogenesis, sperm fertility and infertility, sperm-egg binding, the transfer of sperm components to the fertilized egg, and the functions of sperm components in the early embryo. In more general biological terms, they should provide additional basic information about the movement of plasma membrane proteins in differentiating cells, the relationship between the cytoskeleton and membrane fluidity, membrane fusion, and cellular derepression.