BPA: Cellular and epigenetic effects on the urethra

梗阻性排尿障碍(CVD)是男性衰老过程中的一种常见疾病，虽然不太常见，但AISC 发生在女性身上。拟议的实验涉及出生前和出生后(寿命)暴露的作用。 双酚A(BPA)对雄性和雌性SD大鼠OVD的影响 在NCTR的GLP方案下使用不同剂量的双酚A。男性和女性的初步研究 大鼠和小鼠表明，尿道腺发育和尿路畸形的变化 由雌激素化学物质引起，包括双酚A，但具体效果在低剂量和高剂量下是不同的。 初步研究还表明，出生前暴露于BPA会导致暴露于 成年期的外源性雌激素。我们的假设是胎儿接触低剂量的双酚A将 雄性和雌性SD大鼠在衰老过程中均易发生OVD 在出生后生活中暴露于相同剂量的双酚A。在具体目标1中，我们将检查双酚A的影响 男性和女性胎儿(孕21天)尿道腺结构的研究(计算机辅助 连续切片重建)和上皮和基质的功能，使用激光捕获显微切割 (LCM)结合下一代测序(NGS)用于检测基因表达和表观遗传学 DNA的变化，以及免疫组织化学，以确定特定的细胞类型，增殖和凋亡。 在特定目的2中，将从患OVD或在特定情况下发生OVD的成年雄性和雌性大鼠身上收集尿路 非疾病对照组的年龄。我们将检查尿道腺和间质的结构和功能 对胎儿描述的相同结果(组织病理学、免疫组织化学、LCM捕获上皮 和基质细胞，用NGS分析mRNA和基因组DNA以确定转录谱 并确定基因组DNA的全基因组甲基化图谱。详细信息的组合 不同剂量双酚A对胎儿和成人尿路组织形态和功能的影响 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.