FERTILIZATION DYSFUNCTION AND HUMAN MALE INFERTILITY

大多数男性不育病例都不能得到充分解释，尽管 其中一些被认为与受精障碍有关。我们的目标是 了解细胞和分子生物学的第一个阶段之一 人类受精过程--精子与 卵的透明带-并确定是否有一类雄性 由于这一阶段配子功能障碍而导致的不育症 互动。拟议中的实验将用人类配子来测试一个模型 从动物研究中发展出来的，它表明精子-小带结合是由于 在暴露在外部的质膜精子酶之间形成的键 (半乳糖基转移酶)和N-乙酰氨基葡萄糖末端残基。 透明带。在试点实验中，我们检测到 人精子上的半乳糖基转移酶。在这个项目中，我们将测试 该酶在精子与人透明带结合中的作用 测定：1)酶抑制剂的阻断效果 精子-透明带结合，2)人类透明带作为 精子半乳糖基转移酶催化转移的受体 [~3H]-半乳糖从尿苷二磷[~(3~)]半乳糖和3)的能力 尿苷二磷半乳糖导致与人结合的精子脱落 透明带。不育症患者精子-卵泡结合异常的发生率 将使用一种更精确的分析方法重新调查男性， 精子带结合与精子半乳糖基转移酶的相关性 无论是生育男性还是不育男性，都将进行活跃度测定，以确定是否 精子转移酶活性低于正常可以作为一种指标 不孕不育。最后，人类小带的主要蛋白质成分 用十二烷基硫酸钠/聚丙烯酰胺凝胶电泳法对透明带进行分析。一个 将对人卵透明带溶解制剂进行测试 在人类精子中诱导顶体反应的能力，并观察它是否 含有预期具有精子受体特性的材料。

Most cases of male infertility can not be adequately explained although some are thought to involve fertilization dysfunction. Our objective is to understand the cell and molecular biology of one of the first stages of the human fertilization process - the early association of the sperm with the egg's zona pellucida - and to determine if there is a class of male infertility resulting from dysfunction of this stage of gamete interaction. The proposed experiments will test with human gametes a model developed from animal studies which suggests sperm-zona binding is due to bonds formed between an externally exposed, plasma membrane sperm enzyme (galactosyltransferase) and terminal N-acetylglucosamine residues in the zona pellucida. In pilot experiments, we have detected galactosyltransferase on human sperm. In this project we will test the role of this enzyme in the binding of sperm to the human zona pellucida by determining: 1) the effectiveness of inhibitors of the enzyme in blocking sperm-zona binding, 2) the ability of the human zona pellucida to serve as an acceptor for sperm galactosyltransferase-catalyzed transfer of [3H]-galactose from uridinediphospho[3H]galactose and 3) the ability of uridinediphosphogalactose to cause detachment of sperm bound to the human zona pellucida. The incidence of subnormal sperm-zona binding in infertile men will be reinvestigated using an assay of increased precision, and the correlation between the sperm-zona binding and sperm galactosyltransferase activity will be determined in both fertile and infertile men, to see if subnormal sperm transferase activity can be used as an indicator of infertility. Finally, the major protein components of the human zona pellucida will be analyzed by SDS/polyacrylamide gel electrophoresis. A solubilized preparation of human zonae pellucidae will be tested for its ability to induce acrosome reactions in human sperm and to see if it contains material with properties expected of a sperm receptor.