BPA: Cellular and epigenetic effects on the urethra

描述（由申请人提供）：阻塞性排尿障碍（CVD）是男性在衰老过程中常见的疾病，虽然不常见，但也发生在女性中。在美国国家毒理学研究中心（NCTR）的GLP方案下，研究了双酚A （BPA）在雌雄SD大鼠中产前和产后（寿命）暴露对OVD发展的影响。对雄性和雌性大鼠和小鼠的初步研究表明，包括双酚a在内的雌激素化学物质可引起尿道周围腺发育的变化和尿道畸形，但低剂量和高剂量的具体影响不同。初步研究还表明，产前暴露于BPA会导致成年期暴露于外源性雌激素的雄性小鼠出现OVD。假设胎儿暴露于低剂量双酚a会使雄性和雌性SD大鼠在出生后继续暴露于相同剂量的双酚a中，从而在衰老过程中易患OVD。在目标1中，研究人员将研究BPA对男性和女性胎儿（妊娠第21天）尿道周围腺结构（使用计算机辅助重建系列切片）和上皮和间质功能的影响，使用激光捕获显微解剖（LCM）结合下一代测序（NGS）检查基因表达和DNA表观遗传变化，以及免疫组织化学（IHC）来识别特定的细胞类型，增殖和凋亡。在目标2中，将从患有OVD的成年雄性和雌性大鼠或在特定年龄的非疾病对照中收集尿道。研究人员将使用与胎儿相同的结果（组织病理学、免疫组织化学、LCM）检查尿道周围腺和间质结构和功能，捕获上皮和间质细胞，使用NGS分析mRNA和基因组DNA，以确定转录谱，并确定基因组DNA的全基因组甲基化谱。结合胎儿和成人尿道组织暴露于不同剂量BPA的详细形态和功能分析是非常创新的。

DESCRIPTION (provided by applicant): Obstructive voiding disorder (CVD) is a common disease during aging in men, and while less common, it also occurs in women. The proposed experiments concern the role of prenatal and postnatal (lifespan) exposure to bisphenol A (BPA) on the development of OVD in both male and female SD rats reared and treated with different doses of BPA under a GLP protocol at the National Center for Toxicological Research (NCTR). Preliminary studies with both male and female rats and mice show that changes in periurethral gland development and malformations of the urethra are caused by estrogenic chemicals, including BPA, but specific effects are different at low and high doses. Preliminary studies also show that prenatal exposure to BPA results in OVD in male mice exposed to exogenous estrogen during adulthood. The hypothesis is that fetal exposure to low doses of BPA will predispose both male and female SD rats to development of OVD during aging as a result of continued exposure to the same dose of BPA during postnatal life. In aim 1 the investigators will examine the effects of BPA on male and female fetal (gestation day 21) periurethral gland structure (using computer-assisted reconstruction of serial sections) and function of epithelium and stroma, using laser capture microdissection (LCM) coupled with next generation sequencing (NGS) for examination of gene expression and epigenetic changes in DNA, as well as immunohistochemistry (IHC) to identify specific cell types, proliferation and apoptosis. In aim 2 the urethra will be collected from adult male and female rats that develop OVD or at specific ages in non-diseased controls. The investigators will examine periurethral gland and stroma structure and function using the same outcomes described for fetuses (histopathology, immunohistochemistry, LCM) to capture epithelial and stromal cells for analysis of mRNA and genomic DNA using NGS to determine transcriptomal profiles and to determine genome-wide methylation profiles of genomic DNA. The combination of detailed morphological and functional analyses of fetal and adult urethral tissues exposed to different doses of BPA is highly innovative.