BPA: Cellular and epigenetic effects on the urethra

BPA：对尿道的细胞和表观遗传影响

• 批准号: 8477193
• 负责人: FREDERICK S VOM SAAL
• 金额: $ 18.5万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-19 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8477193
• 关键词: 2 year old; Actins; Acute Renal Failure with Renal Papillary Necrosis; Adult; Affect; Age; Aging; Animal Model; Animals; Apoptosis; Area; Basal Cell; Bioinformatics; Bladder; Breeding; Candidate Disease Gene; Cessation of life; Chemicals; Clinical; Collagen; Computer Assisted; Computers; Coupled; Coupling; Cytokeratin-8 Staining Method; DNA; DNA Methylation; Data; Decision Making; Development; Disease; Dose; Dysplasia; Elastin; Environmental Pollution; Epigenetic Process; Epithelial; Epithelial Cells; Epithelium; Estradiol; Estrogens; Etiology; Exhibits; Exposure to; Female; Fetus; Frequencies; Gene Expression; Genital system; Genomics; Gland; Goals; Growth; Histopathology; Hyperplasia; Immunohistochemistry; In Situ Nick-End Labeling; Instruction; Kinetics; Life; Longevity; MSMB gene; Maps; Messenger RNA; Methods; Molecular Profiling; Morphology; Mus; Neonatal; Outcome; Perinatal Exposure; Positioning Attribute; Pregnancy; Process; Prostate; Prostatic; Prostatic Diseases; Protocols documentation; Rattus; Research; Research Personnel; Role; Serum; Staining method; Stains; Stromal Cells; Structure; Symptoms; Tissues; Urethra; Urethral Obstruction; Urinary tract; Urination; Urine; Urogenital Sinus; Vimentin; Woman; base; bisphenol A; caspase-3; cell type; computerized tools; constriction; fetal; genome wide methylation; gland development; innovation; laser capture microdissection; mRNA Expression; male; malformation; men; next generation sequencing; postnatal; pregnant; prenatal; prenatal exposure; pressure; probasin; programs; protein expression; reconstruction; research study; response; urinary bladder neck

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.

阻塞性空隙障碍（CVD）是男性衰老期间的一种常见疾病，虽然不常见 发生在女性中。提出的实验涉及产前和产后（寿命）暴露的作用 养育和治疗的男性和雌性SD大鼠的Bisphenol A（BPA） 在NCTR处的GLP方案下，BPA的不同剂量。男性和女性的初步研究 大鼠和小鼠表明，尿道周围的腺体发育和尿道畸形的变化是 由包括BPA在内的雌激素化学物质引起的，但在低剂量和高剂量下的特定作用是不同的。 初步研究还表明，暴露于BPA的产前暴露在暴露于雄性小鼠的OVD中 成年期外源性雌激素。我们的假设是，胎儿暴露于低剂量的BPA将会 由于继续 在产后生活期间暴露于同一剂量的BPA。在特定目标1中，我们将检查BPA的影响 在雌性和女性胎儿（妊娠第21天）周围腺体周围结构（使用计算机辅助 串行切片的重建）和上皮和基质的功能，使用激光捕获微分解 （LCM）与下一代测序（NGS）结合，用于检查基因表达和表观遗传学 DNA的变化以及免疫组织化学，以鉴定特定的细胞类型，增殖和凋亡。 在特定的目标2中，尿道将是从成年男性和雌性大鼠中收集的 未解决的对照中的年龄。我们将使用尿道周围的腺体和基质结构和功能使用 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 使用NGS来分析mRNA和基因组DNA的基质细胞，以确定转录谱 并确定基因组DNA的全基因组甲基化谱。详细的结合 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是 高度创新。