FERTILIZATION DYSFUNCTION AND HUMAN MALE INFERTILITY

受精功能障碍和人类男性不育症

基本信息

批准号：
3316967
负责人：
NICHOLAS L CROSS
金额：
$ 9.85万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-09-30 至 1989-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3316967
关键词：
N acetylglucosamine acrosome antibody cell membrane chemical binding egg /ovum electron microscopy fertility fertilization galactosyltransferases gel electrophoresis human subject immunofluorescence technique male sperm sperm capacitation sugar nucleotide

项目摘要

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.

尽管大多数男性不育病例无法得到充分解释有些被认为与受精功能障碍有关。我们的目标是了解细胞和分子生物学的第一阶段之一人类受精过程——精子与受精卵的早期结合卵子的透明带 - 并确定是否有一类雄性此阶段配子功能障碍导致的不孕相互作用。拟议的实验将用人类配子测试模型动物研究表明精子与透明带的结合是由于外部暴露的质膜精子酶之间形成的键（半乳糖基转移酶）和末端 N-乙酰葡糖胺残基透明带。在试点实验中，我们发现人类精子上的半乳糖基转移酶。在这个项目中我们将测试这种酶在精子与人类透明带结合中的作用确定：1) 酶抑制剂阻断的有效性精子-透明带结合，2) 人类透明带充当精子半乳糖基转移酶催化转移的受体 [3H]-半乳糖来自尿苷二磷酸[3H]半乳糖和3)的能力尿苷二磷酸半乳糖导致与人类结合的精子脱离透明带。不育症患者精子带结合异常的发生率将使用更精确的检测方法对男性进行重新调查，并且精子带结合与精子半乳糖基转移酶之间的相关性将测定生育男性和不育男性的活性，看看是否精子转移酶活性低于正常水平可作为以下指标：不孕症。最后，人体透明带的主要蛋白质成分透明菌将通过 SDS/聚丙烯酰胺凝胶电泳进行分析。一个人类透明带溶解制剂将进行测试能够在人类精子中诱导顶体反应并观察它是否含有具有精子受体预期特性的物质。