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

该提案的目的是确定产前环境暴露的机制 内分泌干扰物双酚 A (BPA) 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 使用跨代育种检查 Balb/c 小鼠的产前 BPA（口服）和 PAH（吸入）暴露情况 可以在 F1 后代和 F2 孙代中检查效果的设计。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）以及这些组织的关联 分子变化与神经生物学和生理学结果。整个妊娠期都会发生暴露 产后母婴互动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、家庭笼养 社交互动（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，如果接触 BPA，则需要额外接触 将包括小组以检查免疫功能。暴露引起的脑细胞结构变化将 在 PND 40 时对 F1 和 F2 后代进行量化。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。分子的选择 目标将基于从哥伦比亚儿童中心收集的组织的分析结果 环境健康 (CCCEH) 队列特别关注参与神经发育和 肥胖。甲基化分析将涉及对亚硫酸氢盐处理的 DNA 样品和 mRNA 进行焦磷酸测序 通过定量 RT-PCR 实现分析。总的来说，这些研究旨在确认和验证 CCCEH 人类 BPA 和 PAH 暴露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大脑和脂肪组织中确定的表观遗传变化之间的假设联系 确定介导暴露的长期影响的可能机制途径。

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.