Project 3: Molecular/Disease Consequences of Prenatal BPA, PAH Expos. Across Gene

本提案的目的是确定产前暴露于环境污染的机制， 内分泌干扰物双酚A和多环芳烃的相关水平 多环芳烃对神经生物学、新陈代谢、免疫功能和行为产生长期影响。我们建议 使用跨代繁殖检查Balb/c小鼠产前暴露于BPA（口服）和PAH（吸入） 设计，其中可以在F1后代和F2子代中检查效果。该提案旨在 确定产前BPA和PAH暴露对DNA甲基化和基因表达的不同影响， 几种组织（海马、下丘脑、大脑皮层、脂肪细胞和血液）以及这些组织之间的联系 分子变化与神经生物学和生理学结果。暴露将发生在整个妊娠期 产后母婴相互作用将被评估为对后代的潜在调节影响 发展对雄性和雌性F1和F2后代的评估将包括体重监测、家庭饲养 社交（PND 30-40）、焦虑样行为（PND 40）和认知功能（PND 44-60）。 肥胖将在行为测试后进行量化，在BPA暴露的情况下，额外暴露 将包括各组以检查免疫功能。暴露引起的脑细胞结构的变化将 在PND 40时在F1和F2后代中定量。将评估驱动这些效应的分子机制 在胎儿发育期间（GD 19）和成年期（PND 60）的后代组织中。分子选择 目标将基于从哥伦比亚儿童中心收集的组织分析结果。 环境健康（CCCEH）队列，特别关注参与神经发育和 肥胖甲基化分析将涉及亚硫酸氢盐处理的DNA样品与mRNA的焦磷酸测序 通过定量RT-PCR进行分析。总体而言，这些研究旨在确认和验证 在CCCEH BPA和PAH暴露的人体研究中确定的生物标志物，并确定 血液中的表观遗传变化与脑和脂肪组织中确定的表观遗传变化之间的假设联系， 确定可能的机制途径，通过该途径介导长期暴露效应。

The objective of this proposal is to determine the mechanism by which prenatal exposure to environmentally relevant levels of the endocrine disrupting chemicals bisphenol A (BPA) and polycyclic aromatic hydrocarbons (PAHs) exert long-term effects on neurobiology, metabolism, immune function and behavior. We propose to examine prenatal exposure to BPA (oral) and PAH (inhaled) in Balb/c mice using a transgenerational breeding design in which the effects can be examined in Fl offspring and F2 grand-offspring. This proposal intends to determine the distinct effects of prenatal BPA and PAH exposure on DNA methylation and gene expression in several tissues (hippocampus, hypothalamus, brain cortex, adipocytes and blood) and the association of these molecular changes with neurobiological and physiological outcomes. Exposure will occur throughout gestation and postnatal mother-infant interactions will be assessed as a potential modulating influence on offspring development. Assessment of male and female Fl and F2 offspring will include weight monitoring, homecage social interactions (PND 30-40), anxiety-like behavior (PND 40), and cognitive functioning (PND 44-60). Adiposity will be quantified following behavioral testing and in the case of BPA exposure, additional exposure groups will be included to examine immune function. Exposure-induced changes in brain cytoarchitecture will be quantified in Fl and F2 offspring at PND 40. The molecular mechanisms driving these effects will be assessed in offspring tissue during fetal development (GD 19) and in adulthood (PND 60). The selection of molecular targets will be based on the results of analysis of tissue collected from the Columbia Center for Children's Environmental Health (CCCEH) cohort with particular focus on genes involved in neurodevelopment and obesity. Methylation analysis will involve pyrosequencing of bisulphite treated DNA samples with mRNA analysis achieved through quantitative RT-PCR. Overall, these studies are designed to confirm and validate the biomarkers determined in the CCCEH human studies of BPA and PAH exposure and to determine the hypothesized link between epigenetic changes in blood with those determined in brain and adipose tissue to determine the possible mechanistic pathways through which long-term effects of exposure are mediated.