FUNCTION OF MACAQUE SPERM PH 20 IN FERTILIZATION

猕猴精子 PH 20 在受精中的功能

• 批准号: 6116656
• 负责人: JAMES W OVERSTREET
• 金额: $ 5.91万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6116656
• 关键词: animal tissue; enzymes; male; proteins; reproductive system

Significance The PH-20 protein is one of several molecules on the surface of mammalian sperm which interact with the oocyte investments prior to fertilization. PH-20 appears to have multiple functions and is a target for contraceptive vaccines that are now being developed and tested for effectiveness in primates. Objectives In this project we are studying the cellular mechanisms of action of PH-20 during primate fertilization because it is applicable to mammalian fertilization biology and will be particularly useful in the design of contraceptive vaccines with multi determinant immunogens. Results PH-20 is a hyaluronidase and a requirement for PH-20 function during sperm penetration of the cumulus extra cellular matrix (ECM) is highly likely. Two primary isoforms of macaque sperm PH -20 can be distinguished by their molecular weights of 64 kDa and 53 kDa. Both of these forms of PH-20 are found in three separate locations or compartments of the sperm. Treatment of sperm with hyaluronic acid (HA) results in phosphorylation of sperm proteins, and an increase in intracellular calcium. These HA-induced changes can be inhibited by treatment with Fab fragments of anti-PH-20 IgG. We can show increases in sperm calcium as measured in individual cells during penetration of the macaque cumulus ECM in vitro. The role of HA in mediating such intracellular calcium changes is suggested by similar observations on individual sperm in artificial gels containing HA. The biological importance of intracellular calcium changes may be to "prime" sperm for immediate induction of the acros ome reaction as soon as sperm binding to the zona pellucida takes place. PH-20 may have a required function in fertilization, making it a vulnerable target for contraceptive agents. Future Directions Determine structures of macaque PH-20 isoforms, characterize hyaluronidase and HA binding activities and investigate signaling functions during interaction with the cumulus ECM and zona pellucida. KEY WORDS sperm, fertilization, contraception FUNDING NIH Grant RR00169, HD29125; Andrew Mellon Foundation

显着性pH-20蛋白是该分子的几个分子之一 与卵母细胞投资相互作用的哺乳动物精子的表面 在受精之前。 PH-20似乎具有多个功能，并且 是现在开发的避孕疫苗的目标 并测试了灵长类动物的有效性。 这个项目的目标 我们正在研究pH-20的细胞作用机理 灵长类动物受精是因为它适用于哺乳动物 受精生物学，将在设计中特别有用 具有多种决定因子免疫原的避孕疫苗。 结果 pH-20是透明质酸酶，对PH-20功能的要求 积云额外的细胞基质（ECM）的精子穿透高度 可能。 两种主要的猕猴精子PH -20的同工型可以是 其分子量为64 kDa和53 kDa。 两个都 这些形式的PH-20在三个单独的位置或 精子的车厢。 用透明质酸治疗精子 （HA）导致精子蛋白的磷酸化，并增加 细胞内钙。 这些HA引起的变化可以被 用抗PH-20 IgG的Fab片段处理。 我们可以显示增加 在精子钙中，在渗透过程中在单个细胞中测量 猕猴在体外。 HA在介导的作用 细胞内钙的变化是通过对类似观察到的 含有HA的人造凝胶中的个体精子。 生物学 细胞内钙变化的重要性可能是“ Prime”精子 为了立即诱导Acros OME反应，一旦精子 与Zona Pellucida结合。 ph-20可能需要 受精的功能，使其成为脆弱的目标 避孕剂。 未来的方向决定了结构 猕猴pH-20同工型，表征透明质酸酶和HA结合 活动并研究与 Cumulus ECM和Zona Pellucida。 关键词精子，受精， 避孕资金NIH Grant RR00169，HD29125;安德鲁·梅隆 基础