STUDIES OF SPERM FUNCTION IN MAMMALIAN FERTILIZATION

哺乳动物受精过程中精子功能的研究

• 批准号: 2025037
• 负责人: PATRICIA J OLDS-CLARKE
• 金额: $ 22.66万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-12-01 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2025037
• 关键词: SDS polyacrylamide gel electrophoresis; acrosome; cell membrane; developmental genetics; fertility promoting drug; fertilization; fluorescent dye /probe; gene mutation; genetic mapping; laboratory mouse; molecular genetics; phosphorylation; protein tyrosine kinase; receptor binding; sperm; sperm capacitation; sperm motility; statistics /biometry; western blottings; zona pellucida glycoproteins

DESCRIPTION (Adapted from the Investigator's Abstract): The t haplotypes of mouse sperm are a valuable model system for understanding the genetic and biochemical basis of mammalian sperm dysfunction. Sperm from mice carrying two t haplotypes have extremely poor sperm quality, are deficient in the ability to bind to the zona pellucida, and are unable to penetrate zona-free oocytes. The ultimate objectives of this research are to determine the biochemical mechanisms by which loci in the t haplotypes cause dysfunction in fertilization and to identify and characterize these loci. Gene sequences can then be used as probes to identify human homologs. The sterility of t/t males is caused by the interaction of three genetic factors, S1-S3, each located in a different chromosomal inversion. When homozygous, S1 enhances defects in motility and egg penetration caused by heterozygosity of S2. Aim I is to determine the type of mechanism underlying this enhancement and map the genes responsible to a region small enough for positional cloning. This can be done using Mus spretus-Mus domesticus chromosome 17 recombinant mice, since M. spretus contains noncomplementing alleles to the t haplotype loci affecting sperm, and M. spretus DNA is not inverted relative to M. domesticus. Biochemical defects associated with either S1 or S2 could be involved in zona-binding deficiency. Aim II is to determine which possibility is correct by mapping this deficiency within the t complex. These experiments have the potential to identify the molecular basis for reduced sperm-zona binding. The t haplotype causes an abnormality in the tyrosine phosphorylation of a specific subset of sperm proteins. Since these phosphorylation events are coupled with capacitation in wildtype sperm, the t haplotype-specific defect(s) that affect this pathway could be involved in pathways leading to defective motility and/or egg penetration. Aim III is to determine whether this is so by determining the requirements for the abnormal protein tyrosine phosphorylation, and by mapping the defect to a specific region of the t complex.

描述（改编自研究者的摘要）：t 单倍型 小鼠精子是了解遗传和遗传的一个有价值的模型系统 哺乳动物精子功能障碍的生化基础。 小鼠携带的精子 两个 t 单倍型的精子质量极差，缺乏 能够与透明带结合，并且无法穿透无透明带 卵母细胞。 这项研究的最终目标是确定 t 单倍型中的基因座导致功能障碍的生化机制 受精并识别和表征这些基因座。 基因 然后可以使用序列作为探针来鉴定人类同源物。 t/t雄性不育是由三种遗传相互作用引起的 S1-S3 因子，各自位于不同的染色体倒位。 什么时候 纯合子，S1 增强由以下原因引起的运动和卵子穿透缺陷 S2的杂合性。 目标 I 是确定机制的类型 在这种增强的基础上，绘制负责一个小区域的基因 足以用于定位克隆。 这可以使用 Mus spretus-Mus 来完成 Domesticus 17 号染色体重组小鼠，因为 M. spretus 含有 影响精子的 t 单倍型基因座的非互补等位基因，以及 M. spretus DNA 相对于 M. Domesticus 来说不是倒置的。 与 S1 或 S2 相关的生化缺陷可能涉及 透明带结合缺陷。 目标 II 是确定哪种可能性 通过在 t 复合体中映射此缺陷来纠正。 这些实验 有潜力确定精子带减少的分子基础 绑定。 t单倍型导致酪氨酸异常 精子蛋白特定子集的磷酸化。 由于这些 磷酸化事件与野生型精子的获能相结合， 影响该途径的单倍型特异性缺陷可能涉及 导致运动性和/或卵子穿透缺陷的途径。 目标三是 通过确定以下要求来确定是否如此 异常的蛋白质酪氨酸磷酸化，并通过将缺陷映射到 t复合体的特定区域。