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

PH-20蛋白是细胞膜上的几种分子之一， 哺乳动物精子表面与卵母细胞投资相互作用 在受精之前。 PH-20似乎具有多种功能， 是目前正在开发的避孕疫苗的目标 并在灵长类动物中进行了有效性测试。 本项目的目标 我们正在研究PH-20的细胞作用机制， 灵长类受精，因为它适用于哺乳动物 施肥生物学，将是特别有用的设计， 具有多决定簇免疫原的避孕疫苗。 结果 PH-20是一种透明质酸酶，并且在治疗期间需要PH-20功能。 精子穿透卵丘细胞外基质（ECM）的能力很强， 很有可能。 猕猴精子PH-20的两种主要同种型可以是 它们的分子量分别为64 kDa和53 kDa。 两 这些形式的PH-20在三个不同的地方被发现， 精子的隔间。 透明质酸治疗精子 (HA)导致精子蛋白磷酸化， 胞内钙 这些HA诱导的变化可以被抑制， 用抗PH-20 IgG的Fab片段处理。 我们可以看到 在精子渗透过程中在单个细胞中测量的精子钙 猕猴卵丘细胞外基质的体外培养。 医管局在调解这类 细胞内钙的变化，建议由类似的观察， 含有HA的人工凝胶中的单个精子。 生物 细胞内钙变化的重要性可能是“启动”精子 一旦精子出现， 与透明质酸结合。 PH-20可能需要 在受精中起作用，使其成为一个脆弱的目标， 避孕药。 未来的方向 猕猴PH-20亚型，表征透明质酸酶和HA结合 活动和研究信号功能， 积云ECM和透明积云。 关键词精子，受精， 避孕基金NIH Grant RR 00169，HD29125; Andrew Mellon 基础