MALE REPRODUCTIVE TOXICITY AND ESTROGENICITY OF PAHS

描述：(改编自调查人员摘要)这一目标 研究建议是检验多环芳烃的假设 碳氢化合物(PAHs)对男性生殖系统有害。它是 假设多环芳烃可能作为抗雌激素、雌激素或直接 对生精上皮细胞的毒物。特定目标A：至 鉴定和量化生精上皮细胞的破坏 性成熟雄性大鼠性接触多环芳烃。假设是多环芳烃 引起睾丸细胞结构紊乱扰乱最终发育 成熟的精子。将检查特定亚类的雄性生殖细胞 用形态计量学方法评价组织学制剂的损伤 分析。这项研究将包括酶和细胞表面标记分析， 可能会由于多环芳烃的毒性而改变。特定目标B：确定 性成熟雄鱼多环芳烃影响精子产生的方式， 精子储存和精子形态。假设多环芳烃会改变 睾丸、附睾腺和雄性附性腺重量和组织学 反过来，抑制睾丸精子的产生和储存。量化 对睾丸和附睾精子进行显微镜分析将使用 血细胞计数计。附睾精子的形态也将被评估。特定目标 C：研究多环芳烃暴露对生殖细胞-支持细胞黏附的影响 使用功能生物测定。假设是多环芳烃改变细胞黏附 分子，因此，支持细胞和生殖细胞的相互作用将是无效的。一个 贴壁细胞分析和分类激光扫描细胞仪将测量 支持细胞-生殖细胞黏附。特定目的D：确定 多环芳烃在生精植物特定细胞间连接上的暴露 上皮组织。假设是多环芳烃的暴露扰乱了 相邻支持细胞之间的细胞间连接。紧密连接和缝隙 我们将检查交叉点。将在体内和体内检查连接点。 使用共聚焦分析的体外光学显微镜和超微结构 使用示踪剂研究的观察将包括在内。Western blotting和 定量的酶联免疫吸附试验方法将与针对TECH的抗体一起使用。 结点和缝隙结点。特异目的E：确定多环芳烃是否有毒性 这种作用是通过多环芳烃的雌激素活性来实现的。假设是多环芳烃 暴露是由雌激素受体介导的机制造成的。剂量 不同剂量对MCF-7细胞增殖的影响 将建立细胞所暴露的各种多环芳烃的组合。 雌二醇-17β将被用作比较的标准。这些措施的结果 调查将使我们深入了解 多环芳烃的哪些环境污染物会干扰雄性 生殖系统。

DESCRIPTION: (Adapted from the Investigator's Abstract) The goal of this research proposal is to test the hypothesis that polycyclic aromatic hydrocarbons (PAHs) are toxic to the male reproductive system. It is hypothesized that PAHs may act as anti-estrogens, estrogens or a direct toxicant to the cells of the seminiferous epithelium. SPECIFIC AIM A: To identify and quantify cellular disruption of the seminiferous epithelium in sexually mature male rats due to exposure of PAHs. The hypothesis is that PAHs cause testicular cytoarchitectural disruption perturbing final development of mature spermatozoa. Particular subclasses of male germ cells will be examined to assess the damage from the histological preparations by using morphometric analysis. The study will include enzyme and cell surface marker analysis which may possibly be altered due to PAH toxicity. SPECIFIC AIM B: To determine in sexually mature males the manner in which PAHs influence sperm production, sperm storage, and sperm morphology. The hypothesis is that PAHs alter testicular, epididymal and male accessory sex gland weights and histology, and in turn, suppress testicular sperm production and storage. Quantitative microscopical analysis will be done on testicular and epididymal sperm using a hemocytometer. Epididymal sperm morphology will also be evaluated. SPECIFIC AIM C: To determine the effects of PAHs exposure on germ cell-Sertoli cell adhesion using functional bioassays. The hypothesis is that PAHs alter cell adhesion molecules and thus, Sertoli cell-germ cell interaction will be ineffective. An adherent cell analysis and sorting laser scanning cytometry will measure Sertoli cell-germ cell adhesion. SPECIFIC AIM D: To determine the effects of PAHs exposure on specific intercellular junctions of the seminiferous epithelium. The hypothesis is that PAHs exposure disrupts the formation of intercellular junctions between adjacent Sertoli cells. Tight junctions and gap junctions will be examined. Junctions will be examined both in vivo and in vitro Light microscopy using confocal analysis, and ultrastructural observations using tracer studies will be included. Western blotting and quantitative ELISA methods will be used with antibodies specific for tight junctions and gap junctions. SPECIFIC AIM E: To determine if PAH-induced toxic effect is mediated by estrogenicity of the PAHs. The hypothesis is that PAH exposure results from an estrogen receptor mediated mechanism. The dose response relationship between the proliferation of MCF- 7 cells and the amount of various PAHs to which the cells are exposed will be established. Estradiol-17beta will be used as standard for comparison . Results of these investigations will provide insight into our understanding of the manner in which the PAH environmental pollutants act to interfere with the male reproductive system.