TOXICOLOGICAL EFFECTS OF POLYCHLORINATED HYDROCARBONS ON SPERMATOGENESIS

在这个研究提案中，我们将检验生殖的假设 多氯芳烃(PAHs)对男性的毒性 对睾丸细胞有直接影响。具体目标A：确定 多环芳烃对支持细胞和生(生)精细胞的体外作用 假设多环芳烃导致两种支持细胞存活率下降 细胞和生殖细胞。我们将在孤立的环境中进行这些实验 培养的细胞。将检查细胞的形态和功能。流动 将采用细胞分析技术。这项研究将包括支持细胞 细胞骨架组织和分泌功能可能是 由于多环芳烃的毒性而改变。特异目的B：测定体内 多环芳烃对生精上皮细胞的影响他们 假设暴露于多环芳烃会导致睾丸破裂。 导致分化生殖细胞数量减少的细胞结构。 我们将对暴露在体内的睾丸进行光学和电子显微镜检查 多环芳烃，并将使用形态测量法量化损害。生殖细胞将是 用流式细胞仪进行定量。特异目的C：研究黄连素的作用 腐殖质对多环芳烃急性毒性的影响假设是富里奇 酸、腐植酸和胡敏素可用作多环芳烃毒性的处理。 分离的支持细胞和生殖细胞。我们将做实验来证明 这些细胞在含有商品的培养液中孵育 在随后添加多环芳烃之前的腐殖质会阻止 存活率下降，细胞功能下降。特定目标D：学习 多环芳烃诱导Sertoli细胞毒性的可能机制 细胞和生殖细胞。这一假说将用DNA进行检验 3‘-羟基DNA末端的电泳法和ApopTag标记。具体目标E：TO 确定多环芳烃引起的睾丸细胞改变 岩藻糖基转移酶(FT)活性，支持细胞和生殖细胞不 能够维持必要的体细胞-生发细胞黏附。生物检测 将对FT活性进行量化，激光扫描细胞仪将测量 支持细胞-生殖细胞黏附。这项调查的结果将 让我们深入了解多环芳烃 环境污染物会干扰精子的发生。

In this research proposal we will test the hypothesis that reproductive toxicity in the male by polychlorinated aromatic hydrocarbons (PAHs) involves direct effects on testicular cells. SPECIFIC AIM A: To determine in vitro effects of PAHs on Sertoli cells and spermatogenic (germ) cells. The hypothesis is that PAHs cause decreased viability of both Sertoli cells and germ cells. We will conduct these experiments in isolated cultured cells. Cell morphology and functions will be examined. Flow cytometry analysis will be employed. The study will include Sertoli cell cytoskeleton organization and secretory functions which may possibly be altered due to PAHs toxicity. SPECIFIC AIM B: To determine the in vivo effects of PAHs on the cells of the seminiferous epithelium. They hypothesis is that exposure to PAHs will cause disruption of testicular cytoarchitecture yielding decreased number of differentiating germ cells. We will perform light and electron microscopy of testes exposed in vivo to PAHs and will quantify damage using morphometry. Germ cells will be quantified using flow cytometry. SPECIFIC AIM C: To study the effects of humic substances on acute toxicity of PAHs. The hypothesis is that fulvic acid, humic acid, and humin may be used as treatments in PAHs toxicity on isolated Sertoli cells and germ cells. We will do experiments to show that incubation of these cells in medium containing commercially available humic substances prior to the subsequent addition of PAHs block the decreased viability and decreased cell function. SPECIFIC AIM D: To study the possible apoptic mechanisms in PAHs-induced cytotoxicity in Sertoli cells and germ cells. The hypothesis will be tested using DNA electrophoresis and ApopTag labeling of 3'-OH DNA ends. Specific Aim E: To determine that PAHs induced changes in testicular cells alter fucosyltransferase (FT) activity, and Sertoli cells and germ cells are not able to maintain necessary somatic cell-germinal cell adhesion. Bioassays will quantify FT activity and a laser scanning cytometry will measure Sertoli cell-germ cell adhesion. Results from this investigation will provide insight into our understanding of the manner in which PAHs environmental pollutants act to interfere with spermatogenesis.