BPA: Cellular and epigenetic effects on the urethra

阻塞性排尿障碍（CVD）是男性衰老过程中的常见疾病，虽然不太常见，但也是一种常见疾病。 发生在女性身上。拟议的实验涉及产前和产后（寿命）暴露的作用 双酚A（BPA）对雄性和雌性SD大鼠OVD发展的影响， 在NCTR的GLP协议下使用不同剂量的BPA。男性和女性的初步研究 大鼠和小鼠表明，尿道周围腺发育的变化和尿道畸形是 这是由雌激素化学物质引起的，包括BPA，但在低剂量和高剂量下的具体影响不同。 初步研究还表明，产前暴露于BPA会导致雄性小鼠的OVD， 外源性雌激素我们的假设是，胎儿暴露于低剂量的BPA， 在衰老过程中，雄性和雌性SD大鼠由于持续的 在出生后的生活中暴露于相同剂量的BPA。在具体目标1中，我们将研究BPA的影响 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 连续切片的重建）和上皮和间质的功能，使用激光捕获显微切割 (LCM)结合下一代测序（NGS），用于检查基因表达和表观遗传 DNA的变化，以及免疫组化，以确定特定的细胞类型，增殖和凋亡。 在特定目标2中，将从发生OVD的成年雄性和雌性大鼠或在特定时间采集尿道。 未患病对照组的年龄。我们将检查尿道周围腺和基质的结构和功能， 胎儿的结局相同（组织病理学、免疫组织化学、LCM捕获上皮细胞 和基质细胞，用于使用NGS分析mRNA和基因组DNA以确定转录组谱 并确定基因组DNA的全基因组甲基化谱。详细的组合 暴露于不同剂量BPA的胎儿和成人尿道组织的形态和功能分析， 极具创新性。

Obstructive voiding disorder (CVD) is a common disease during aging in men, and while less common, it aisc 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 NCTR. Preliminary studies with both male and female rats and mice show that changes in periurethral gland development and malformations ofthe 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. Our 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 specific aim 1 we 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 to identify specific cell types, proliferation and apoptosis. In Specific Aim 2 the urethra will be collected from adult male and female rats that develop OVD or at specific ages in non-diseased controls. We 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.