MAMMALIAN BOMBESIN-LIKE PEPTIDES IN SPERM FERTILIZATION

精子受精中的哺乳动物铃蟾肽样肽

• 批准号: 2202578
• 负责人: ELIOT R SPINDEL
• 金额: $ 15.14万
• 依托单位: OREGON REGIONAL PRIMATE RESEARCH CENTER
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-12-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2202578
• 关键词: Macaca mulatta; Xenopus oocyte; acidity /alkalinity; acrosome; bombesin; calcium channel blockers; calcium flux; fertilization; human tissue; immunocytochemistry; in situ hybridization; in vitro fertilization; laboratory rabbit; mass spectrometry; molecular cloning; neuropeptide receptor; peptide hormone biosynthesis; peptide structure; pertussis toxin; protein sequence; radioimmunoassay; receptor expression; sperm; sperm capacitation; sperm motility

Fertilization of mammalian ovum by sperm is a complicated multistep process involving binding of sperm to the egg, induction of the acrosome reaction (an exocytic activation of the sperm) and penetration of the sperm through the zona pellucida. Our laboratory has made the new observation that bombesin-like peptides appear to be endogenous regulators of the primate acrosome reaction. The bombesin-like peptides are a unique, wide-ranging family of peptides originally found in frog skin and later characterized in mammals. The mammalian bombesin-like peptides, of which gastrin-releasing peptide (GRP) is the best characterized member, are important neurotransmitters, growth factors, and paracrine hormones. The role of GRP as a potential regulator of the growth of small cell lung carcinoma has been widely studied. Previously our laboratory has characterized the genes and cDNAs encoding GRP and the GRP receptor. We have made the new observation that a splice variant of the GRP mRNA is expressed in primate testes and encodes a new bombesin-like peptide that is capable of inducing the acrosome reaction. Preliminary studies in vitro have shown that 1) bombesin-like peptides stimulate the acrosome reaction in both rhesus monkey and human sperm; 2) that specific GRP receptor antagonists block this stimulation and also block the progesterone-induced acrosome reaction in monkey and human sperm, and 3) that GRP antagonists inhibit the fertilization of monkey oocytes by sperm in in vitro fertilization (IVF) assays. The purpose of this grant is to characterize the role of bombesin-like peptides in the acrosome reaction and fertilization of primate ova by sperm. Specifically, we propose: 1) to characterize the endogenous bombesin-like peptide(s) which induce the acrosome reaction; 2) to characterize the bombesin-like peptide receptor expressed in sperm; 3) to characterize in vitro the conditions, time course and mechanisms by which bombesin agonists stimulate and bombesin antagonists inhibit the acrosome reaction in sperm; and 4) to investigate the potential role of bombesin- like peptides in fertilization using in vitro fertilization of monkey oocytes by monkey sperm. The bombesin-like peptides and receptors will be characterized by molecular and conventional techniques. Studies on the role of bombesin-like peptides on fertilization will take advantage of the ongoing rhesus monkey IVF program at the Oregon Regional Primate Research Center. Thus, the aim of this research is to characterize a new intermediate step in the fertilization of ova by sperm and to generate potential new approaches to problems of human fertility and contraception.

精子对哺乳动物卵的施肥是复杂的多步 涉及精子与鸡蛋结合的过程，诱导缩小型 反应（精子的胞外激活）和渗透 精子穿过Zona Pellucida。我们的实验室使新的 观察到类似孟买的肽似乎是内源性调节剂 灵长类动物的顶体反应。 类似孟买的肽是一个独特的，广泛的肽家族 最初在青蛙皮肤中发现，后来以哺乳动物为特征。这 哺乳动物类似的bombesin样肽，其中胃蛋白释放肽（GRP） 是最有特征的成员，是重要的神经递质，生长 因素和旁分泌激素。 GRP作为潜在调节剂的作用 小细胞肺癌的生长已被广泛研究。 以前，我们的实验室已经表征了基因和cDNA编码 GRP和GRP受体。我们已经做出了新的观察结果 GRP mRNA的变体在灵长类睾丸中表达并编码一个新的 能够诱导顶体反应的bombesin样肽。 体外的初步研究表明，1）bombesin样肽 刺激恒河猴和人类精子中的Adrosom反应； 2） 特定的GRP受体拮抗剂阻止了这种刺激，也阻止 阻止孕激素诱导的猴子和人类的Astrosom反应 精子和3）GRP拮抗剂抑制猴子的受精 精子在体外受精（IVF）测定中的卵母细胞。 这笔赠款的目的是表征类似孟买的角色 肽中的肽和灵长类动物OVA受精的肽 精子。 具体来说，我们建议：1）表征内源性 轰炸型肽（S），诱导了Adrosom反应； 2）到 表征精子中表达的孟买样肽受体； 3）到 体外表征条件，时间过程和机制 孟买激动剂刺激和轰炸的拮抗剂抑制了ACROSOME 精子中的反应； 4）调查孟买的潜在作用 例如使用猴子的体外受精在受精中的肽 猴子精子的卵母细胞。类似孟买的肽和受体将是 以分子和常规技术为特征。研究 类似孟买的肽在受精方面的作用将利用 俄勒冈州地区灵长类动物研究的恒河猴IVF计划正在进行 中心。 因此，这项研究的目的是表征一个新的中间步骤 精子的卵子受精并产生潜在的新 解决人类生育和避孕问题的方法。