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: 3316225
• 负责人: DAVID J NISHIOKA
• 金额: $ 6.08万
• 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/3316225
• 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 精子表面在精子发生过程中的分化以及精子 受精后卵表面成分分散。 更具体的目标包括：（1）使用单克隆抗体 针对位点特异性精子表面抗原，以确定何时在 精子发生和在睾丸中这些抗原被添加到 生精细胞表面;（2）克隆这些基因的编码 位点特异性表面抗原，并且使用这些克隆的基因，（3） 确定在精子发生过程中何时以及在睾丸中的何处， 基因转录;（4）准备和筛选合适的表达 可以从中分离这些基因的文库，（5）亚克隆这些 基因导入合适的转录载体，以及（6）设计 有效的切片技术， 杂交和抗体结合研究;（7）确定是否 位点特异性精子表面抗原横向分散在表面 多精受精卵，如果是这样，（8）以确定 细胞骨架和代谢抑制剂对这一横向的影响 传播;以及，使用最近开发的融合鸡蛋的程序，（9） 以确定精子表面抗原是否均匀地分散在未受精的 受精卵融合和受精卵-受精卵融合。 由于将要研究的表面抗原具有Ca 2+通道 能力和已知参与精子顶体反应 和精卵结合，这些目标和决定应该提供 更深入地了解精子发生，精子生育力和 不孕症，精卵结合，这些目标和决定应该 更深入地了解精子发生、精子生育力和 不育，精卵结合，精子成分转移到 受精卵和精子成分在早期胚胎发育中的作用 胚胎 在更一般的生物学术语中，它们应该提供 关于质膜运动的附加基本信息 分化细胞中的蛋白质， 细胞骨架和膜流动性、膜融合和细胞 去压抑