Electrogenic bicarbonate secretion in Calu-3 cells

Calu-3 细胞中的电动碳酸氢盐分泌

• 批准号: 6800850
• 负责人: JAMES Levin KREINDLER
• 金额: $ 5.44万
• 依托单位: CHILDREN'S HOSP PITTSBURGH/UPMC HLTH SYS
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-12 至 2005-08-11
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6800850
• 关键词: RNA; bicarbonates; cell line; chloride channels; cystic fibrosis; immunocytochemistry; immunoprecipitation; ion transport; membrane; mucus; polymerase chain reaction; postdoctoral investigator; respiratory epithelium; transfection /expression vector

DESCRIPTION (provided by applicant): Abnormal bicarbonate secretion has long been recognized as a key component of cystic fibrosis (CF) pancreatic dysfunction, but has only recently been investigated as a component of CF lung disease. The submucoal glands of the lungs are responsible for secretion of a complex airway surface liquid (ASL) that lines the airway epithelium and plays a significant role in mucociliary clearance. The serous cell is the predominant cell type of the submucosal gland and the predominant site of cystic fibrosis transmembrane conductance regulator (CFTR) expression. The Calu-3 cell, an immortalized serous cell line, is therefore a good model for the study of the physiology of ASL production and regulation. This proposal is divided into two complementary aims. The first aim is to use microelectrode measurements to establish the transport physiology of bicarbonate entry on a sodium-bicarbonate cotransporter, and then to use short, inhibiting RNA technology to determine the identity of the sodium-bicarbonate cotransporter. The second aim is to use short, inhibitory RNA technology to knock-down CFTR and to use these knock-down cell lines in Ussing chamber experiments and microelectrode experiments to test the hypothesis that CFTR is the apical bicarbonate channel.

描述（由申请人提供）：长期以来，异常的碳酸氢盐分泌已被认为是囊性纤维化（CF）胰腺功能障碍的关键组成部分，但直到最近才被研究为CF肺部疾病的组成部分。肺的粘膜粘膜负责分泌一个复杂的气道表面液体（ASL），该液体（ASL）在气道上皮上排列并在粘膜纤毛间隙中起重要作用。浆液细胞是粘膜粘膜的主要细胞类型，是囊性纤维化跨膜电导调节剂（CFTR）表达的主要位点。因此，Calu-3细胞是一种永生的浆液细胞系，是研究ASL生产和调节生理学的好模型。该提议分为两个补充目标。第一个目的是使用微电极测量值来在二氯甲酸钠共转运蛋白上建立碳酸氢盐进入的运输生理，然后使用短而抑制的RNA技术来确定二碳酸盐盐碳酸盐盐的身份。第二个目的是使用简短的抑制性RNA技术来敲除CFTR，并在USSING室实验中使用这些敲低的细胞系和微电极实验来测试CFTR是碳酸氢盐通道的假设。