DIOXIN BINDING EFFECTS ON OVARIAN HISTOLOGY & MODULATION OF STEROIDOGENESIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的：探讨2，3，7，8-四氯二苯并对二恶英对猕猴卵巢碎片的培养及其对卵泡数量和大小的影响。 关于有毒的二恶英(TCDD)对非人类灵长类动物生殖功能的影响的研究一直很有限。我们已经证明了在恒河猴卵巢组织中存在能够与DNA结合的功能性TCDD或芳香烃(AH)受体，并定位了这种结合到卵泡，并报道了与TCDD相关的性类固醇在培养中通过卵巢碎片产生的改变。在后一种模型系统中，我们现在描述TCDD暴露对卵泡大小和数量的影响。三个新鲜的猕猴卵巢(猕猴；WNPRC；威斯康星州麦迪逊)被清除多余的组织，冲洗(PBS)，称重并切成碎片。将碎片放置在单独的Falcon器官培养皿中，在37℃，5%CO2，98%增湿空气中培养在含有二氧六环(CTL)、皮摩尔(PM)、纳米摩尔(NM)或微摩尔(？M)浓度的TCDD的培养液(无酚红DMEM/F12，FBS，50ug/ml庆大霉素)中培养96h。所有碎片称重，冷冻(-80℃)，冷冻(10？M)，苏木精-伊红染色。用光学显微镜对卵泡大小进行量化。尽管动物之间存在差异，但分析表明，与对照碎片相比，暴露于TCDD的碎片的卵泡总数往往减少(%CTL；？M：78%；NM：57%；PM：65%)，特别是在中等大小的碎片(最大横截面积为104？106M2；%CTL；？M：68%；NM：44%；PM：59%)。我们希望用这种方法作为研究环境污染物对猴子生育能力影响的模型。这个项目使用了WNPRC病理服务。 该项目得到了LKC的ASP和WNPRC的NIH P51 RR000167、RR15459-01、RR020141-01的部分支持。资金在本报告所述期间之前结束，但出版物尚待出版。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Objective: To evaluate the culture of rhesus monkey ovarian fragments with 2,3,7,8-tetrachlorodibenzo-p-dioxin and assess effects on follicle number and size. There has been limited investigation of the effects of the toxicant dioxin (TCDD) on the reproductive function of non-human primates. We have demonstrated the presence of functional TCDD- or aromatic hydrocarbon (AH)-receptor capable of binding DNA in rhesus monkey ovarian tissue, localized that binding to follicles, and reported TCDD-related altered production of sex steroids by ovarian fragments in culture. We now describe the effects of TCDD exposure on ovarian follicle size and number in the latter model system. Three fresh monkey ovaries (Macaca mulatta; WNPRC; Madison, WI) were cleaned of excess tissue, rinsed (PBS), weighed and quartered into fragments. Fragments were placed in separate Falcon organ culture dishes and incubated (37¿C, 5% CO2, 98% humidified air) in culture medium (phenol red-free DMEM/F12, FBS, 50 ug/ml gentamycin) containing either dioxane (CTL), or picomolar (pM), nanomolar (nM), or micromolar (¿M) concentrations of TCDD for up to 96 hours. All fragments were weighed, stored frozen (-80 ¿C), cryosectioned (10 ¿M) and stained with hematoxylin and eosin. Quantification of follicles by size was performed using light microscopy. Although variability was noted between animals, analyses show that fragments exposed to TCDD tended to have a decreased total number of follicles when compared to control fragments (%CTL; ¿M:78%; nM:57%; pM:65%), especially in the mid-sized range (greatest cross sectional area of 104  106 ¿M2 ;%CTL; ¿M:68%; nM:44%; pM:59%). We hope to use this approach as a model for studying environmental contaminants on monkey fertility. This project used WNPRC Pathology Services. This project was supported, in part, by ASP to LKC, and NIH P51 RR000167, RR15459-01, RR020141-01 to WNPRC. Funding ended before this reporting period but publications are pending.