Ubiquitin-Based Semen Qualtiy Assay in Toxicology

接触工作场所化学品的男性的精液质量是 NIOSH计划公告的优先事项#RFA-OH-00-006。这项建议是 旨在评估工作场所化学品对精液的有害影响 男性工作者的质量，使用一种新的、客观的、非侵入性的精液分析， 并致力于建立一种相关的、非人类的灵长类动物生殖模型 毒物学。目前还没有提供客观的精液质量分析方法 工人的精液筛查是基于临床精子参数(活动率， 计数、形态)，通过高度主观的形态评估进行评估，或者 充其量，通过计算机化的精子运动分析。 我以前已经指出，选择性泛素化，即标记 精子线粒体膜上含有通用蛋白降解标志物多肽， 泛素存在于睾丸和受精后(Sutovsky等人，1999， 自然402：371-372)。我的新研究表明，由于这个最初的睾丸 泛素化，所有源于细胞的有缺陷的精子和精液污染物 在附睾期内变得泛素化。的相对水平 用流式细胞仪、酶联免疫吸附试验和放射免疫法可以客观地测定精液中的泛素 免疫荧光法使用抗泛素抗体，并反映生育能力 人类和动物的繁殖性能。因此，泛素似乎是一种 精液异常的通用生物标志物。 这一建议解决了以下问题：1.是否检测到 精子表面泛素化--评估精液质量的通用方法 在接触工人中？2.恒河猴是最好的动物模型吗 评估工作场所化学品对男性生育能力和精液的影响 质量？如果完全开发，这个项目将允许快速、大规模 对接触工人的精液样本进行筛选，并建立恒河猴 猴子作为人类生殖毒理学的相关模型。这个项目将 通过提供第一个真正的目标来对公共卫生产生积极影响 和职业医学通用精液质量检测。

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.