BIOCHEMICAL MARKERS OF SPERM FERTILITY

精子生育能力的生化标志物

• 批准号: 2206189
• 负责人: GABOR B HUSZAR
• 金额: $ 18.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-05-05 至 2000-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2206189
• 关键词: SDS polyacrylamide gel electrophoresis; acrosome; biomarker; chromatin; creatine kinase; egg /ovum; enzyme activity; fertility; fertility immunology; fertilization; gel electrophoresis; high performance liquid chromatography; histones; human subject; hyaluronate; ion exchange chromatography; protein isoforms; semen; sperm; sperm capacitation

he central theme of our research is idiopathic or unexplained male infertility which affects about 1.5 million American men. For the present proposal we have identified a fertIle control population. and five groups of men with idiopathic male infertility: (a) Husbands of Infertile couples referred to our Sperm Physiology Laboratory (about 650 men/1000 samples per year); (b) Unexplained male infertility (already identified normospermic men with diminished sperm CK-M ratios); (c) Fertile and infertile patients with varicocele; (d) Husbands of 1300 infertile couples in which a detailed data base was developed, including their reproductive history. lifestyle, drug use and other epidemiological characteristics; and (e) Patients who receive medications that affect sperm function. In order to develop improved methods for the diagnosis and treatment of men with idiopathic infertility in the physician's office and in the Andrology laboratory we propose to study: 1) Sperm creatine kinase (CK) activity and CK-M isoform ratios, objective biochemical markers of sperm cellular maturity and fertilizing potential. Within this aim, we will: (a) Isolate the CK-M isoform of human sperm and will generate CK-M specific antibodies for the assessment of maturity rather than morphology of ejaculated sperm. (b) Develop sperm CK antibodies which will not react with seminal fluid CK, in order to directly measure sperm Ck in semen samples by immunotechnology already present in clinical laboratories and in doctors' offices. 2) In line with our hypothesis, which assumes that idiopathic infertility may be caused by isolated deficiencies of acrosome activation in mature sperm, and that in vitro enhancement of sperm function may correct this problem, we will (a) study the rate of both capacitation and acrosome reaction in fertile men and in men with idiopathic infertility; (b) study acrosome activation and the CK parameters in the same sample and (c) examine the possible correlation between sperm CK parameters and acrosome function in the paradigms of the hamster oocyte penetration assay and (ARIC). the Ca- ionophore mediated acrosome response test. 3) Following up on our discovery that hyaluronic acid increases sperm velocity, we will: (a) Study sperm velocity response in a hyaluronic acid "challenge test"; (b) Compare, in a kit available in doctors' offices, sperm progress In cervical mucus with and without hyaluronic acid; and (c) Develop hyaluronic acid beads for the selection of mature, viable sperm for intracytoplasmic injection. 4) Because idiopathic male infertility may arise due to various pathomechanisms, we will compare the diagnostic efficiency of sperm CK-parameters with "strict" sperm morphology, with anilin blue staining, which probes sperm chromatin maturity, and with the hyaluronic acid velocity response test.

我们研究的中心主题是特发性或不明原因的男性 影响大约150万美国男性的不孕症。目前而言 建议我们已经确定了一个有生育能力的控制人口。和五个小组 患有特发性男性不育的男性：(A)不育的丈夫 夫妇向我们的精子生理学实验室咨询(约650名男性/1000人 (B)原因不明的男性不育(已查明 精子CK-M比率降低的正常精子男性)；(C)有生育能力和 精索静脉曲张不育患者；(D)1300名不育患者的丈夫 开发了详细的数据库的夫妇，包括他们的 生育史。生活方式、吸毒和其他流行病学 特征；和(E)接受药物治疗的患者 精子功能。为了开发改进的诊断方法 和治疗男性特发性不孕症的医生 办公室和男科实验室，我们计划研究：1)精子 肌酸激酶(CK)活性和CK-M亚型比率，目标 精子细胞成熟度和受精潜能的生化标志物。 在这一目标中，我们将：(A)分离人类精子的CK-M亚型，并 会产生CK-M特异性抗体来评估成熟度 而不是射出精子的形态。(B)发育精子CK 不会与精液CK反应的抗体，以便 已通过免疫技术直接检测精液样本中的精子CK 出现在临床实验室和医生办公室。2)符合 我们的假设，假设特发性不孕症可能导致 由于成熟精子的顶体激活的孤立缺陷，以及 在体外增强精子功能可能会纠正这个问题，我们将 (A)研究生育能力和顶体反应的比率 男性和患有特发性不育症的男性；(B)研究顶体激活 和同一样本中的CK参数，以及(C)检查可能的 精子CK参数与精子顶体功能的相关性 仓鼠卵母细胞穿透试验和(ARIC)的范例。钙-- 离子载体介导的顶体反应试验。3)跟进我们的 发现透明质酸提高精子速度，我们将：(A) 研究透明质酸“挑战试验”中的精子速度反应；(B) 在医生办公室提供的试剂盒中，比较精子的进展 宫颈粘液含有和不含透明质酸；和(C) 透明质酸珠用于选择成熟的、可存活的精子 胞浆内注射。4)因为特发性男性不育可能 由于各种发病机制引起的，我们将比较诊断 精子CK参数与“严格的”精子形态的效率 苯胺蓝染色，用于探测精子染色质成熟度，并与 透明质酸速度反应试验。