BIOCHEMISTRY OF SPERM-EGG INTERACTION AND FERTILIZATION

精卵相互作用和受精的生物化学

• 批准号: 3312047
• 负责人: VICTOR D VACQUIER
• 金额: $ 15.25万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-06-01 至 1987-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3312047
• 关键词: acrosome; affinity chromatography; calcium; cell adhesion; cell membrane; egg /ovum; fertilization; germ cells; methylation; peptidases; phosphorylation; saltwater environment; sperm

The long-term objective of this research program is to gain a detailed biochemical knowledge of the process of animal fertilization. This proposals describes basic research that will increase our knowledge of processes underlying sperm-egg interaction during fertilization. This knowledge is essential to the future development of non-hormonal methods of contraception in higher forms such as man. Sea urchins are used in this work because they are the only group of animals producing enough sperm nd eggs to permit biochemical studies. What little we do know about mammalian fertilization shows that striking similarities exist between mammals and sea urchins thus making the work on sea urchins relevant to mammals. The sea urchin is the paradigm for fertilization research just as the mouse is for cancer research. The specific aims of this program are: to characterize and determine the function of three sperm membrane proteins which we call 84K, 110K & 115K. We believe 84K is involved in triggering the sperm acrosome necessary for binding the sperm to the egg. The 110K & 115K together comprise 50% of the sperm membrane protein, so they must be very important to the sperm function. We can isolate the sperm membrane in the form of vesicles (SMV) which we will characterize fully and also study their interaction with eggs. We will determine if methylation and phosphorylation of sperm lipids and proteins occurs during the acrosome reaction. We will isolate and characterize the sperm surface antigenic component common to all animal sperm we have tested. This may have the potential of being an ideal immunocontraceptive. We will study the sperm binding protein, bindin, in greater detail and determine the amino acid sequence of two bindins and also study its carbohydrate binding specificity. We will study the mechanism by which extracellular calcium activates the adenylate cyclase of abalone sperm, and the mechanism by which sea urchin eggs redistribute bindin-containing vesicles on their surfaces. We will determine the role of the corticsal granule glucanase in fertilization and also identify specific phosphoproteins regulating the metabolic state of the egg before, during and after fertilization. Lastly, we will study the biochemistry of the calcium-mediated explosion of isolated egg cortical granuels. The success of this project will greatly increase our knowledge of sperm-egg interaction.

该研究计划的长期目标是获得详细的 动物受精过程的生化知识。 这 提案描述了将增加我们知识的基础研究 受精过程中精卵相互作用的过程。 这 知识对于非激素方法的未来发展至关重要 更高形式的避孕，例如男性。 这个用的是海胆 工作，因为它们是唯一产生足够精子的动物群体 鸡蛋以进行生化研究。 我们对哺乳动物知之甚少 受精表明哺乳动物和人类之间存在惊人的相似之处 因此，海胆研究与哺乳动物相关。 这 海胆是受精研究的典范，就像老鼠一样 用于癌症研究。 该计划的具体目标是： 表征并确定三种精子膜蛋白的功能 我们称之为 84K、110K 和 115K。 我们相信 84K 参与触发 将精子与卵子结合所必需的精子顶体。 110K & 115K 共同构成了精子膜蛋白的 50%，因此它们必须是 对精子功能非常重要。 我们可以分离精子膜 我们将充分表征并研究囊泡（SMV）的形式 他们与鸡蛋的相互作用。 我们将确定是否甲基化和 精子脂质和蛋白质的磷酸化发生在顶体期间 反应。 我们将分离并表征精子表面抗原 我们测试过的所有动物精子共有的成分。 这可能有 具有成为理想免疫避孕药的潜力。 我们将研究精子 结合蛋白，结合蛋白，更详细地确定氨基酸 两个结合蛋白的序列并研究其碳水化合物结合 特异性。 我们将研究细胞外钙的机制 激活鲍鱼精子的腺苷酸环化酶，其机制是 哪些海胆卵将含有结合蛋白的囊泡重新分布在其上 表面。 我们将确定皮质颗粒葡聚糖酶的作用 在受精过程中，还可以识别调节受精的特定磷蛋白 受精前、受精期间和受精后卵子的代谢状态。 最后， 我们将研究钙介导的爆炸的生物化学 分离的鸡蛋皮质颗粒。 该项目的成功将极大 增加我们对精子与卵子相互作用的了解。