EPIDIDYMAL MATURATION OF MAMMALIAN SPERM

这项研究计划的广泛和长期目标是了解 精子的附睾成熟过程。 这个定义不清的过程 是受精所必需的精子进入尾部 附睾和输精管在获能一段时间后， 使卵子受精，而从睾丸或附睾头取出的精子 没有 工作模型是精子在附睾内的成熟是 不是一个单一的事件，而是涉及精子特性的顺序，渐进 由附睾分泌物的信号传导引起 精子的细胞内空间。 理解的挑战 因此，精子在附睾中的成熟是决定性因素之一， 对受精所必需的精子成分进行修改， 描述这些组件如何因以下原因而发挥功能： 附睾诱导的改变，并阐明附睾因素或 刺激这些变化的条件。 这项研究将 检查在重要精子的范围内发生的变化， 顶体，顶体。 需要调查的变更是 精子特异性丝氨酸的结构和功能修饰 蛋白酶酶原、顶体酶原和另一个顶体的结构变化 糖蛋白，顶粒蛋白。 具体目标1将使用高 碳水化合物分析的高效液相色谱法 发生在体中的顶体蛋白原寡糖的修饰 附睾 具体目标2将确定这些变化如何影响 酶活性 特异性目标3将确定特异性裂解 在顶粒蛋白的多肽骨架中由部分氨基酸 从附睾尾部精子分离的产物的测序。 具体目标 4将使用特定目标1和3的生化信息， 预测和鉴定特定的糖苷酶和蛋白酶 负责附睾诱导的顶体蛋白原的改变， 顶粒蛋白 具体目标5是开发一种生物测定， 作为附睾标记物的顶体蛋白原和顶粒蛋白的修饰 成熟 该生物测定将用于确定条件或 引起睾丸和附睾头这些变化的物质 精子 这些实验将提供一种独特的方法来研究 精子的附睾成熟。 在拟议的生物测定期间开发的 项目将用于未来的研究，以表征条件或 纯化负责启动精子这方面的因素 成熟 此外，从这些研究中获得的信息将 发现在治疗由阻塞引起的男性不育症中的应用， 附睾或输精管 发展新的 男性避孕药也可能来自这些信息， 问题研究

The broad, long term objective of this research proposal is to understand the process of epididymal maturation of sperm. this poorly defined process is essential for fertilization; sperm that have entered the cauda epididymis and vas deferens are able, after a period of capacitation, to fertilize eggs whereas sperm taken from the testis or caput epididymis are not. The working model is that sperm maturation within the epididymis is not a single event but involves sequential, progressive in sperm properties brought about by the transduction of signals from the epididymal secretions to the intracellular spaces of the sperm. The challenge in understanding sperm maturation in the epididymis, therefore, is one of determining the modifications to sperm components that are essential for fertilization, characterizing how these components become functional as a result of epididymis-induced alterations, and elucidating the epididymal factors or conditions that stimulate these changes. This proposed research will examine changes that take place within the confines of the important sperm acrosome, the acrosome. The alterations to be investigated are the structural and functional modifications of the sperm-specific serine protease zymogen, proacrosin, and structural changes to another acrosomal glycoprotein, acrogranin. specific Aim 1 will characterize, using high performance liquid chromatography for carbohydrate analysis, the modification of proacrosin oligosaccharides that takes place in the corpus epididymis. Specific Aim 2 will determine how these alterations affect enzymatic activity. Specific Aim 3 will determine where specific cleavages in the polypeptide backbone of acrogranin occur by partial amino acid sequencing of products isolated from cauda epididymal sperm. Specific Aim 4 will use the biochemical information from Specific Aims 1 and 3 to predict and to identify the specific glycosidase(s) and protease(s) responsible for the epididymis-induced alterations of proacrosin and acrogranin. Specific Aim 5 is to develop a bioassay using the modifications of proacrosin and acrogranin as markers of epididymal maturation. this bioassay will be used to determine the conditions or substances that initiate these changes in testicular and caput epididymal sperm. These experiments will provide a unique approach to investigate epididymal maturation of sperm. The bioassay developed during the proposed project will be used in future studies to characterize the conditions or purify the factors responsible for initiating this aspect of sperm maturation. Furthermore, the information gained from these studies will find application in the treatment of male infertility caused by blockage of the epididymis or vas deferens. New approaches toward development of a male contraceptive may also result from the information gained from these studies.