Ubiquitin-Based Semen Qualtiy Assay in Toxicology

基于泛素的毒理学精液质量测定

• 批准号: 6314778
• 负责人: PETER Sutovsky SUTOVSKY
• 金额: $ 9.37万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-30 至 2002-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6314778
• 关键词: Macaca mulatta; antiantibody; clinical research; disease /disorder model; environmental toxicology; enzyme linked immunosorbent assay; fertility; flow cytometry; hazardous substances; human subject; male; mitochondrial membrane; model design /development; occupational hazard; semen; ubiquitin; work site

Semen quality in men exposed to workplace chemicals is one of the priorities of NIOSH's Program Announcement #RFA-OH-00-006. This proposal is aimed at assessing the detrimental effects of workplace chemicals on semen quality in male workers, using a novel, objective, non-invasive semen assay, and at establishing a relevant, non-human primate model for reproductive toxicology. Objective semen quality assays are currently not available and semen screening in workers is based on clinical sperm parameters (motility, count, morphology), assessed by highly subjective morphological evaluation, or at best, by computerized sperm motility analysis. I have shown previously that selective ubiquitination, i.e. the tagging of sperm mitochondrial membranes with universal proteolytic marker peptide, ubiquitin, occurs in testis and after fertilization (Sutovsky et al., 1999, Nature 402:371-372). My new studies show that, due to this initial testicular ubiquitination, all defective sperm and semen contaminants of cellular origin become polyubiquitinated during epididymal passage. The relative levels of ubiquitin in semen can be objectively determined by flow cytometry, ELISA and immunofluorescence using anti-ubiquitin antibodies, and reflect the fertility of men and reproductive performance of animals. Thus, ubiquitin appears to be a universal biomarker of semen abnormalities. This proposal addresses the following questions: 1. Is the detection of sperm-surface ubiquitination the universal approach for assessing semen quality in exposed workers? 2. Is the rhesus monkey the optimal animal model for assessing the effects of workplace chemicals on male fertility and semen quality? If fully developed, this project will allow for rapid, large scale screening of semen samples from exposed workers, and establish the rhesus monkey as a relevant model for human reproductive toxicology. This project will have a positive impact on public health by providing the first truly objective and universal semen quality assay for occupational medicine.

暴露于工作场所化学品的男性的精液质量是影响男性健康的因素之一。 NIOSH计划公告#RFA-OH-00-006的优先事项。这项建议是 旨在评估工作场所化学品对精液的有害影响 男性工人的精液质量，使用一种新的，客观的，非侵入性的精液分析， 并建立一个相关的，非人类灵长类动物模型， 毒理学目前还没有客观的精液质量检测方法， 工人的精液筛选基于临床精子参数（活力， 计数，形态学），通过高度主观的形态学评价进行评估，或 充其量，通过计算机化精子活力分析。 我以前已经证明，选择性泛素化，即标记的 精子线粒体膜与通用蛋白水解标记肽， 泛素，发生在睾丸中和受精后（Sutovsky等人，1999, Nature 402：371-372）。我的新研究表明，由于最初的睾丸 泛素化，所有有缺陷的精子和细胞来源的精液污染物 在附睾通过过程中变得多泛素化。的相对水平 流式细胞术、酶联免疫吸附试验（ELISA）和 免疫荧光使用抗泛素抗体，并反映生育能力 和动物的繁殖能力。因此，泛素似乎是一种 精液异常的通用生物标志物。 本建议涉及以下问题：1.是检测 精子表面泛素化是评价精液质量的通用方法 暴露的工人？2.恒河猴是最佳的动物模型吗 评估工作场所化学品对男性生育能力和精液的影响 质量如何？如果充分开发，该项目将允许快速，大规模 筛选精液样本，从接触工人，并建立恒河猴 猴作为人类生殖毒理学的相关模型。该项目将 通过提供第一个真正的目标， 和职业医学通用精液质量检测。