MECHANISMS OF CELL VOLUME REGULATION AND LIVER FUNCTION

细胞体积调节和肝功能的机制

• 批准号: 2441764
• 负责人: RICHARD M ROMAN
• 金额: $ 10.6万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-15 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2441764
• 关键词: CHO cells; Xenopus oocyte; adenosinetriphosphatase; autocrine; bile; cell morphology; chloride channels; cholestasis; homeostasis; human tissue; immunocytochemistry; in situ hybridization; laboratory rat; liver function; membrane channels; membrane transport proteins; molecular cloning; northern blottings; paracrine; polymerase chain reaction; protein kinase C; purinergic receptor; secretion; site directed mutagenesis; voltage /patch clamp; western blottings

DESCRIPTION (taken from application) Preliminary investigations have identified new insights regarding the roles of Cl- channels and cellular ATP release during liver cell swelling, and molecular correlates of the Cl- channels involved in these pathways. The goals of these studies are to better elucidate the cellular and molecular mechanisms involved in liver cell volume regulation, and implications for liver function. The specific aims are to i) characterize channel-mediated ATP efflux and auto-/ paracrine modulation of liver cell volume and bile formation, ii) investigate the role of liver ATP-binding cassette (ABC) proteins as ATP channels or channel regulators, iii) assess the role of protein kinase C (PKC) as a signaling factor that regulates volume recovery, and iv) identify the molecular basis for increased Cl-permeability during swelling and purinergic stimulation. Using both hepatocyte and biliary cells and cell lines, techniques will include a) study of swelling-activated Cl- and ATP channels using patch clamp analysis, b) measurement of cell volume changes with a Coulter Multisizer, c) characterization of epithelial transport with Ussing chamber voltage-clamp experiments in polarized primary rat cholangioctye monolayers and d) cloning and functional characterization of hepatobiliary CLC Cl- channel homologues. Studies will be performed by the applicant at the University of Colorado Health Sciences Center under the preceptorship of Dr. J. Gregory Fitz. During this mentored research period, investigative tools in cell biology, physiology, and molecular biology will be developed to compliment electrophysiologic study of liver ion channels, and serve as a basis for future contributions in the unexplored area of hepatobiliary ion channel genetics. The long term objective of these investigations is to provide a physiologic basis for the development of pharmacological approaches to modulate biliary secretion in cholestatic liver diseases such as cystic fibrosis, and to ameliorate cellular injury during metabolic stress.

描述（取自应用程序） 初步调查发现了有关这些角色的新见解 在肝细胞肿胀期间Cl-通道和细胞ATP释放，以及 分子相关的氯离子通道参与这些途径。 的 这些研究的目的是更好地阐明细胞和分子 参与肝细胞体积调节的机制， 肝功能 具体目标是i）表征通道介导的 肝细胞体积和胆汁的ATP流出和自分泌/旁分泌调节 ii）研究肝ATP结合盒（ABC）的作用 蛋白作为ATP通道或通道调节剂，iii）评估 蛋白激酶C（PKC）作为调节容量恢复的信号传导因子， 和iv）鉴定在施用期间增加Cl-渗透性的分子基础。 肿胀和嘌呤能刺激。 同时使用肝细胞和胆汁 细胞和细胞系，技术将包括a）溶胀激活的 使用膜片钳分析的Cl-和ATP通道，B）细胞的测量 使用Coulter Multisizer的体积变化，c）上皮细胞的表征 在极化初级电极中用Ussing室电压钳进行的输运实验 大鼠胆管细胞单层和d）克隆和功能表征 肝胆CLC Cl-通道同源物。 研究将由 科罗拉多大学健康科学中心的申请人 J·格雷戈里·菲茨博士的指导 在这段指导研究期间， 细胞生物学、生理学和分子生物学的研究工具将 被开发以补充肝离子通道的电生理学研究， 并作为今后在未开发领域作出贡献的基础， 肝胆离子通道遗传学 这些长期目标 研究的目的是为发展提供生理学基础， 胆汁淤积性疾病中调节胆汁分泌的药理学方法 肝脏疾病，如囊性纤维化，并改善细胞损伤 代谢应激时的反应