BIOCHEMISTRY OF SPERM-EGG INTERACTION AND FERTILIZATION

这项研究计划的长期目标是获得详细的 动物受精过程的生物化学知识。 这 建议描述了基础研究，将增加我们的知识， 受精过程中精卵相互作用的基本过程。 这 知识是必不可少的非激素方法的未来发展， 避孕在更高的形式，如男子。海胆被用于这一点， 工作，因为他们是唯一一组动物产生足够的精子， 鸡蛋，以进行生物化学研究。 我们对哺乳动物所知甚少 受精表明哺乳动物和哺乳动物之间存在着惊人的相似之处， 因此，海胆的研究与哺乳动物有关。 的 海胆是受精研究的典范，就像老鼠一样。 用于癌症研究。 该方案的具体目标是： 对三种精子膜蛋白的功能进行了表征和测定 我们称之为84K、110K和115K。 我们认为84K与触发 使精子与卵子结合的精子顶体。 110K & 115K共同构成了精子膜蛋白的50%，所以它们必须是 对精子功能非常重要。 我们可以分离精子膜 囊泡（SMV）的形式，我们将充分表征，并研究 与鸡蛋的互动。 我们将确定甲基化和 精子脂质和蛋白质的磷酸化发生在顶体 反应 我们将分离并鉴定精子表面抗原 我们测试过的所有动物精子的共同成分。 这可以具有如下 有可能成为一种理想的免疫避孕药。 我们将研究精子 结合蛋白，bindin，更详细，并确定氨基酸 序列的两个结合，并研究其糖结合 的特异性 我们将研究细胞外钙离子 激活鲍鱼精子的腺苷酸环化酶， 海胆卵将含有结合素的囊泡重新分配到它们的 表面。 我们将确定皮质颗粒葡聚糖酶的作用 在受精过程中，还确定了特定的磷蛋白， 卵子在受精前、中、后的代谢状态。 最后， 我们将研究钙介导的爆炸的生物化学， 分离的卵皮质颗粒。 这个项目的成功将大大 增加我们对精子和卵子相互作用的了解。

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.