Identification of Molecular Biomarkers in Rat Sperm and Testis Exposed to BPA

双酚A(BPA)是一种内分泌干扰性化学物质，在已开发的 世界。动物研究和最近的流行病学证据都表明，双酚A针对的是男性 生殖道，造成睾丸损伤和精子参数改变。 在啮齿动物模型中，基于GLP的标准方法在检测BPA诱导的BPA方面相对不敏感 男性生殖道异常。我们建议使用成熟的形态测量法 对睾丸的评估，包括保留精子细胞头部的数量和TUNEL阳性的发生率 生殖细胞，以提高检测双酚A暴露对睾丸影响的敏感性。此外，我们 将通过检测全球信使核糖核酸和微小核糖核酸，确定影响睾丸和精子的分子生物标记物。 水平和全基因组DNA甲基化。测量精子中的这些分子生物标记物有望 要特别敏感和特异，因为精子是发育成熟的同质细胞群体 在它们的整个生命历史中，都在生精上皮内。GLP设计提供多种剂量，并且 积极的雌二醇对照，允许对双酚A诱导效应的存在进行强有力的测试。此外，我们 将评估父系FO大鼠及其后代，解决世代之间的差异。目标是 本项目的目的是检验以下工作假设：添加增强的形态和 GLP设计的分子终点将提供更敏感和特异的双酚A诱导的生物标志物 男性生殖毒性。 GLP研究将通过形态计量评估和影响的分子生物标记物来加强 通过以下具体目标： 具体目标#1：将加强对双酚A所致睾丸损伤的评估 组织学和形态计量学终点。 特定目的#2：鉴定尾部精子和睾丸的mRNA和miRNA转录本的含量。 具体目标#3：发育尾部精子和睾丸甲基组。

Bisphenol A (BPA) is an endocrine disrupting chemical causing ubiquitous exposures in the developed world. Both animal studies and recent epidemiological evidence indicate that BPA targets the male reproductive tracl<, producing testicular injury and alterations in sperm parameters. Standard GLP-based approaches in rodent models have been relatively insensitive in detecting BPAinduced abnormalities in the male reproductive tract. We propose the use of well-established morphometric assessments ofthe testis, including quantitation of retained spermatid heads and the incidence of TUNELpositive germ cells, to enhance the sensitivity of detecting testicular effects of BPA exposure. In addition, we will identify molecular biomarkers of effect in the testis and sperm, by examining global mRNA and miRNA levels and genome-wide DNA methylation. Measuring these molecular biomarkers in sperm is expected to be particularly sensitive and specific, since sperm are an homogeneous population of cells that developed throughout their life history within the seminiferous epithelium. The GLP design provides multiple doses, and a positive estradiol control, allowing robust testing for the presence of a BPA-induced effect. In addition, we will assess both the paternal FO rats and their progeny, addressing differences across generations. The goal of this project is to test the following working hypothesis: addition ofthe enhanced morphological and molecular endpoints with the GLP design will provide more sensitive and specific biomarkers of BPA-induced male reproductive toxicity. The GLP study will be enhanced by morphometric assessments and molecular biomarkers of effects through the following Specific Aims: Specific Aim #1: will enhance the assessment of BPA-induced testicular injury using advanced histological and morphometric endpoints. Specific Aim #2: characterize the caudal sperm and testis mRNA and miRNA transcript content. Specific Aim #3: develop the caudal sperm and testis methylome.