THE SPERM SURFACE IN SPERMATOGENESIS AND FERTILIZATION

精子发生和受精过程中的精子表面

• 批准号: 3316220
• 负责人: DAVID J NISHIOKA
• 金额: $ 6.01万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1992-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3316220
• 关键词: acrosome; autoradiography; calcium channel; cell fusion; cell nucleus; cytoplasm; egg /ovum; fertilization; genetic transcription; immunocytochemistry; immunofluorescence technique; in situ hybridization; membrane activity; membrane proteins; molecular cloning; monoclonal antibody; nonmammalian vertebrate embryology; phase contrast microscopy; saltwater environment; sea urchins; spermatogenesis; surface antigens; testis

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.

本研究计划的总体目标是确定如何 精子表面通过精子发生进行分化以及精子如何分化 受精后表面成分分散在卵子表面。 更具体的目标包括：（1）单克隆抗体的使用 针对位点特异性精子表面抗原，以确定何时 精子发生以及睾丸中这些抗原被添加到的位置 生精细胞表面； (2) 克隆编码这些基因的基因 位点特异性表面抗原，并使用这些克隆基因，（3） 确定精子发生过程中的时间以及睾丸中的位置 基因被转录； (4)准备并筛选合适的表达方式 可以从中分离这些基因的文库，(5) 亚克隆这些基因 将基因转化为适当的转录载体，并且（6）设计 有效的切片技术可进行原位确定 杂交和抗体结合研究； (7) 判断是否 位点特异性精子表面抗原横向分散在表面上 多精受精卵的数量，如果是，则 (8) 确定 细胞骨架和代谢抑制剂对该侧向的影响 分散；并且，使用最近开发的融合鸡蛋程序，（9） 确定精子表面抗原是否均匀分散在未受精的细胞中 受精卵和受精卵与受精卵的融合。 由于要研究的表面抗原具有 Ca2+ 通道 能力并已知参与精子顶体反应 和精卵结合，这些目标和决定应提供 更深入地了解精子发生、精子生育力和 不孕症、精卵结合，这些目标和决心应该 提供对精子发生、精子生育力和 不孕症、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。 用更一般的生物学术语来说，它们应该提供 有关质膜运动的其他基本信息 细胞分化过程中的蛋白质之间的关系 细胞骨架和膜流动性、膜融合和细胞 去压抑。