ELECTROLYTE TRANSPORT IN CULTURED HUMAN CERVICAL CELLS

申请人乔治·戈罗德斯基是一名 产科、妇科医生，生殖科学助理教授 凯斯西储大学生物学教授，长期致力于 生殖生理学研究生涯。他过去在类固醇方面的工作 人体子宫在体内的功能产生了几篇论文。在… 目前，他的研究是针对细胞层面的，他的广泛， 长期目标是研究体液和血液循环的机制和调节。 人类女性生殖道中的溶质转运及其在体内的作用 人类生殖与妇女健康。 本研究的具体目的是：1.优化 人宫颈外上皮细胞的培养方法， 这是一个很好的女性下生殖道的体外模型 上皮，作为多孔过滤器上的极化上皮，并 对培养的上皮细胞进行形态、生化和组织化学鉴定 功能标准。2.为了表征电解液的运输 培养的hCEs细胞，特别强调活性氯化钠的分泌和 吸收，以及所涉及的调节机制。假设是 这种上皮细胞通过主动地控制腔内液体环境 将血液中的液体和溶质分泌到管腔，这就是 过程是在类固醇激素和维甲酸的强效控制下进行的 它可以被神经体液分泌激动剂或 局部炎症反应的介质。与健康相关的人 项目是：1.这些数据将提供细胞机制的新知识。 以及女性生殖道内液体分泌的调节。2. 所获得的信息将使设计临床研究成为可能。 局部使用(即流明)调节离子传输的药剂 以控制女性生殖道内的液体分泌。 实验设计为研究跨上皮氯化钠 利用细胞电生理学特性进行运输。要采取的方法是 用于：1.以短路形式定量测定活性Na~+和CI~-转运 Ussing小室中培养的hECEs细胞产生的电流(ISC)。 2.特定条件下主动迁移的离子的性质 将根据运输治疗后ISC的变化确定 缓蚀剂或在洗浴溶液中进行离子置换之后。3.至 非融合培养体系中氯离子转运的定量研究 荧光显微镜。这项研究将在以下机构的赞助下进行 S博士在生理和生物物理系的伤痕和霍费尔 凯斯西储大学生殖生物学专业。

The applicant, George Gorodeski, is an Obstetrician-Gynecologist and an Assistant Professor of Reproductive Biology at Case Western Reserve University with a long-term commitment to research career in Physiology of Reproduction. His past work on steroid function in the human uterus in-vivo generated several publications. At present, his research is directed at the cellular level and his broad, long term objective is to study mechanisms and regulation of fluid and solute transport in the human female reproductive tract and their role in human reproduction and women's health. The specific aims of the present study are: 1. to optimize the methodology of culturing human ectocervical epithelial (hECE) cells, which are a good in-vitro model of the woman's lower genital tract epithelium, as a polarized epithelium on a porous filter and to characterize the cultured epithelium using morphological, biochemical and functional criteria. 2. to characterize electrolyte transport by the cultured hECE cells with particular emphasis on active NaCl secretion and absorption, and the regulatory mechanisms involved. The hypothesis is that this epithelium controls the luminal fluid environment by actively secreting fluid and solutes from the blood into the lumen, that this process is under the tonic control of steroid hormones and retinoids and that it can be acutely modulated by neurohumoral secretagogues or by mediators of local inflammatory response. The health relatedness of the project is: 1. the data will provide new knowledge of cellular mechanisms and regulation of fluid secretion in the woman's reproductive tract. 2. The information gained will make it possible to design clinical studies of the local use (i.e. luminal) of agents which regulate ion transport in order to control fluid secretion in the woman's genital tract. The experimental design is to study transepithelial NaCl transport using cellular electrophysiology properties. The methods to be used are: 1. to quantitate active Na+ and CI- transport as short circuit current (Isc) generated by cultured hECE cells mounted in Ussing chamber. 2. The nature of the actively transported ions under specific conditions will be determined from Isc changes following treatment with transport inhibitors or following ion replacement in the bathing solution. 3. To quantitate Cl- transport in non-confluent hECE cultures using fluorescence microscopy. The study will be done under the sponsorship of Dr. s' Scarpa and Hopfer at the Departments of Physiology and Biophysics and Reproductive Biology at Case Western Reserve University.