ELECTROLYTE TRANSPORT IN CULTURED HUMAN CERVICAL CELLS

申请人乔治戈罗代斯基是一名 妇产科医生和生殖医学助理教授 凯斯西储大学生物学专业，长期致力于 生殖生理学的研究生涯 他过去对类固醇的研究 在人体子宫内的功能产生了几篇出版物。 在 目前，他的研究是针对细胞水平和他的广泛， 长期目标是研究流体的机制和调节， 人类女性生殖道中的溶质转运及其在 人类生殖和妇女健康。 本研究的具体目的是：1.优化 培养人宫颈上皮细胞（hECE）的方法， 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， 使用形态学、生物化学和 功能标准。2.为了表征电解质传输， 培养的hECE细胞，特别强调活性NaCl分泌， 吸收，以及相关的调节机制。 这个假设是 这种上皮细胞通过主动地控制管腔内的液体环境， 从血液中分泌液体和溶质进入管腔， 这一过程受到类固醇激素和类维生素A的紧张性控制， 它可以被神经体液促分泌素或 局部炎症反应的介质。 的健康相关性 项目是：1.这些数据将为细胞机制提供新的知识 以及调节女性生殖道中的液体分泌。2. 获得的信息将使设计临床研究成为可能 局部使用（即管腔）调节离子转运的药物， 以控制女性生殖道中的液体分泌。 实验设计是研究跨上皮NaCl 使用细胞电生理学特性的运输。 该方法的 使用的是：1.以定量作为短路的活性Na+和Cl-转运 电流（Isc）由安装在Ussing室中的培养的hECE细胞产生。 2.在特定条件下主动输运离子的性质 将根据运输处理后的Isc变化确定 抑制剂或随后在浴溶液中进行离子置换。3.到 在非融合的hECE培养物中定量Cl-转运， 荧光显微镜 这项研究将在以下机构的赞助下进行： 博士生理学和生物物理学系的斯卡帕和霍普费尔 凯斯西储大学的生殖生物学

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.