DIOXIN BINDING EFFECTS ON OVARIAN HISTOLOGY & MODULATION OF STEROIDOGENESIS

二恶英结合对卵巢组织学的影响

• 批准号: 7958735
• 负责人: REINHOLD Josef HUTZ
• 金额: $ 4.68万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7958735
• 关键词: Air; Animals; Area; Binding; Biological Models; Carbon Dioxide; Computer Retrieval of Information on Scientific Projects Database; Cryoultramicrotomy; Culture Media; DNA Binding; Dioxanes; Dioxins; Environmental Pollution; Fertility; Freezing; Funding; Gonadal Steroid Hormones; Grant; Hematoxylin and Eosin Staining Method; Histology; Hour; Incubated; Institution; Investigation; Macaca mulatta; Monkeys; Organ Culture Techniques; Ovarian; Ovarian Follicle; Ovarian Tissue; Ovary; Pathology; Phenolsulfonphthalein; Primates; Production; Publications; Reporting; Research; Research Personnel; Resources; Services; Source; Staining method; Stains; Steroid biosynthesis; Study models; Tetrachlorodibenzodioxin; Tissues; United States National Institutes of Health; Wisconsin; aromatic hydrocarbon receptor; light microscopy; nonhuman primate; reproductive function; toxicant

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.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 目的：评价2，3，7，8-四氯二苯并-对-二恶英对恒河猴卵巢碎片的培养及其对卵泡数量和大小的影响。 有毒物质二恶英（TCDD）对非人灵长类动物生殖功能的影响研究有限。 我们已经证明了功能性TCDD-或芳香烃（AH）-受体的存在，能够结合DNA在恒河猴卵巢组织，本地化，结合卵泡，并报告TCDD相关的改变性类固醇的生产卵巢碎片在文化。 我们现在描述在后一个模型系统中TCDD暴露对卵泡大小和数量的影响。 将三个新鲜猴卵巢（Macaca mulatta; WNPRC;麦迪逊，WI）清除多余组织，冲洗（PBS），称重并切成碎片。将片段置于单独的Falcon器官培养皿中，并在含有二氧六环（CTL）或皮摩尔（pM）、纳摩尔（nM）或微摩尔（<$M）浓度的TCDD的培养基（无酚红DMEM/F12、FBS、50 μ g/ml庆大霉素）中孵育（37 ℃、5% CO2、98%潮湿空气）长达96小时。 对所有片段称重，冷冻储存（-80 ℃），冷冻切片（10 μ M）并用苏木精和伊红染色。 使用光学显微镜对卵泡大小进行定量。尽管动物之间存在差异，但分析表明，与对照片段相比，暴露于TCDD的片段往往具有减少的卵泡总数（%CTL; μ M：78%; nM：57%; pM：65%），特别是在中等大小范围内（最大横截面积为104  106 μ M 2;%CTL; μ 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力的模型。本项目使用WNPRC病理学服务。 本项目部分得到了ASP对LKC和NIH P51 RR 000167、RR 15459 -01、RR 020141 -01对WNPRC的支持。在本报告所述期间之前，供资结束，但出版物尚未出版。