Regulation of Epididymal Luminal Acidification

附睾管腔酸化的调节

基本信息

批准号：
7253878
负责人：
NURIA M. PASTOR-SOLER
金额：
$ 12.96万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-07-01 至 2009-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7253878
关键词：
Acids Acute Address Adenylate Cyclase Adult Affect Agonist Androgen Antagonists Animals Apical Basic Science Bicarbonates Biological Cadmium Carrier Proteins Cell membrane Cells Chronic Clear Cell Condition Cyclic AMP Cyclic AMP-Dependent Protein Kinases Cyclic GMP Data Development Diethylstilbestrol Duct (organ) structure Electrodes Environment Enzymes Epididymis Epithelial Epithelial Cells Estrogens Ethinyl Estradiol Fertility Fluorescence Flutamide GTP-Binding Proteins General Hospitals Goals Gonadotropin Hormone Releasing Hormone Gonadotropin Releasing Hormone Inhibitor Hormonal Hormone Antagonists Hormones In Situ In Vitro Kidney Knowledge Laboratories Lead Massachusetts Mediating Medicine Membrane Biology Modeling Modification Molecular Mouse Protein Mus Neonatal Numbers Organ Phenotype Physiological Play Poison Production Proteins Proton Pump Proton-Translocating ATPases Protons Rattus Recycling Regulation Research Research Proposals Role Second Messenger Systems Site Source Techniques Thinking Toxin Transgenic Mice Vas deferens structure Vesicle Work adenylyl cyclase 6 apical membrane base career cell motility cell type enhanced green fluorescent protein in vivo interdisciplinary approach male mature animal postnatal programs promoter protein expression reproductive research study response second messenger sperm cell steroid hormone toxicant trafficking urologic

项目摘要

DESCRIPTION (provided by applicant): This applicant proposes a program of research to prepare her for a career in academic medicine and basic science research studying male reproductive/urologic tract function, addressing the regulation of epididymal luminal acidification under physiological and pathophysiological conditions, during adulthood and postnatal development. The research will be conducted in the laboratory of Dr. Sylvie Breton at the Program in Membrane Biology and Renal Unit, Massachusetts General Hospital. The epididymis and vas deferens (VD) are the sites where spermatozoa mature and are stored in an immotile state. The luminal environment of these organs is tightly regulated and maintained at an acidic pH in order to keep spermatozoa from achieving motility. In particular, the applicant will study the mechanism by which an alkaline epididymal/VD luminal pH stimulates H+ATPase trafficking from subapical vesicles to the apical plasma membrane in clear cells. In addition, she will further investigate the role of the bicarbonate-activated "soluble" adenylyl cyclase (sAC), cAMP, and downstream effectors (such as PKA and Epac) on this trafficking phenomenon. Other sources of cAMP, such as G protein-regulated transmembrane adenylyl cyclases, and other second messenger molecules (such as cGMP) in clear cells will also be investigated. These studies will also evaluate the effects of steroid hormone agonists and antagonists (DES, ethinyl estradiol, GnRH antagonist and flutamide) and toxic compounds (Cadmium and Lead) have on the epididymal cellular profile, protein expression, H+ATPase trafficking and function, and on the cellular profile of the epididymis/VD during adulthood. The work proposed in this application will be carried out on rat and/or mouse epididymal/VD epithelial cells during adulthood and postnatal development, in vivo, in situ, and in vitro, using a multidisciplinary approach to evaluate protein expression and vesicle recycling mechanisms. The techniques proposed include microspectrofluorescence, cell biological, molecular biological techniques, and the use of a proton-selective self-referencing electrode. Newly available transgenic mice expressing enhanced green fluorescence protein (EGFP) specifically in clear cells (B1-EGFP mice) will provide a unique model to examine the development and modulation of epithelial cell phenotypes in response to various manipulation.

描述（由申请人提供）：该申请人提出了一项研究计划，为她从事学术医学和基础科学研究做好准备，研究男性生殖/泌尿道功能，解决成年期间生理和病理生理条件下附睾管腔酸化的调节问题和产后发育。该研究将在马萨诸塞州总医院膜生物学和肾脏科项目的 Sylvie Breton 博士的实验室中进行。附睾和输精管（VD）是精子成熟并以不动状态储存的场所。这些器官的管腔环境受到严格调节并维持在酸性 pH 值下，以防止精子获得活力。特别地，申请人将研究碱性附睾/VD管腔pH刺激H+ATP酶从顶端下囊泡运输至透明细胞顶端质膜的机制。此外，她还将进一步研究碳酸氢盐激活的“可溶性”腺苷酸环化酶（sAC）、cAMP和下游效应器（如PKA和Epac）对这种贩运现象的作用。还将研究透明细胞中 cAMP 的其他来源，例如 G 蛋白调节的跨膜腺苷酸环化酶，以及其他第二信使分子（例如 cGMP）。这些研究还将评估类固醇激素激动剂和拮抗剂（DES、乙炔雌二醇、GnRH 拮抗剂和氟他胺）和有毒化合物（镉和铅）对附睾细胞特征、蛋白质表达、H+ATP 酶运输和功能的影响，以及成年期附睾/VD 的细胞特征。本申请中提出的工作将在成年和产后发育期间的大鼠和/或小鼠附睾/VD上皮细胞上进行，体内、原位和体外，使用多学科方法评估蛋白质表达和囊泡回收机制。提出的技术包括显微分光荧光、细胞生物学、分子生物学技术以及质子选择性自参考电极的使用。新推出的在透明细胞中特异性表达增强型绿色荧光蛋白 (EGFP) 的转基因小鼠（B1-EGFP 小鼠）将提供一个独特的模型来检查上皮细胞表型响应各种操作的发育和调节。