MECHANISMS OF CELL VOLUME REGULATION AND LIVER FUNCTION

细胞体积调节和肝功能的机制

• 批准号: 6124728
• 负责人: RICHARD M ROMAN
• 金额: $ 11.9万
• 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 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6124728
• 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) 初步调查发现了关于这些角色的新见解 肝细胞肿胀时氯离子通道和细胞ATP释放的变化，以及 参与这些途径的氯离子通道的分子相关性。这个 这些研究的目标是更好地阐明细胞和分子 肝细胞体积调节的机制及其对 肝功能。具体目标是：i)描述渠道中介 肝细胞体积和胆汁的三磷酸腺苷外流和自/旁分泌调节 形成，II)研究肝脏ATP结合盒(ABC)的作用 蛋白质作为ATP通道或通道调节器，III)评估 蛋白激酶C(PKC)作为调节容量恢复的信号因子， 以及iv)确定氯离子渗透性增加的分子基础 肿胀和嘌呤能刺激。肝细胞与胆汁并用 细胞和细胞系，技术将包括)肿胀激活的研究 用膜片钳分析CL-和ATP通道，b)细胞测量 Coulter Multisizer的体积变化，c)上皮细胞的特征 偏振光传输的电压室钳制实验 大鼠胆管单层及d)克隆和功能鉴定 肝胆ClC氯离子通道同系物。研究将由以下人员进行 科罗拉多大学健康科学中心的申请者 J·格雷戈里·菲茨博士的教职。在这一指导研究期间， 细胞生物学、生理学和分子生物学的研究工具将 是对肝脏离子通道的电生理研究的补充， 并作为未来对未勘探区域的贡献的基础 肝胆离子通道遗传学。这些措施的长期目标是 调查是为了提供一个生理基础的发展 胆汁淤积症中调节胆汁分泌的药理学途径 肝病，如囊性纤维化，并减轻细胞损伤 在新陈代谢压力下。