Water Transport in the Lacrimal Gland

泪腺中的水运输

• 批准号: 6598689
• 负责人: PETER R BRINK
• 金额: $ 30.1万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6598689
• 关键词: apical membrane; basolateral membrane; biological fluid transport; chloride channels; enzyme activity; enzyme induction /repression; gene targeting; genetically modified animals; immunocytochemistry; laboratory mouse; lacrimal apparatus; membrane transport proteins; potassium channel; protein kinase C; secretion; sodium channel; voltage /patch clamp; voltage gated channel; water channel; western blottings

DESCRIPTION (provided by applicant): Fluid secreted by the lacrimal gland is an essential component of tears and is estimated to contribute approximately 50% of the liquid volume bathing the cornea (Walcott, 1998). Recently in situ measurements of fluid production from the lacrimal glands of mice have demonstrated stimulated flow rates of 0.2-0.6 uL/minute (Walcott et al., 2002; Paranyuk et al., 2001; Moore et al, 2000). Our preliminary data indicates that the NKCC1 Na+, K+, 2CI- co-transporter and Bkca channels play important roles in fluid production of the lacrimal gland. We propose the use of knockout mice along with NZB mice (compromised fluid flow, Paranyuk et al., 2001) to study their respective roles in fluid production by the lacrimal gland. Further we propose the use of activators and inhibitors of PKC to delineate its role in the regulation of the NKCC1 co-transporter and Bkca channel. Both systems have been shown to be influenced/moduate/regulate by PKC (Standen and Quayle, 1998;Zhou et al., 2001; Clerice etal., 1995). Our proposal is focused on addressing the following hypotheses: Aim 1 Hypothesis: Basolateral blockade of the salt co-transporter (NKCC1) will significantly reduce stimulated fluid flow in controls but will be similar to NKCC1 knockout flow rates. We will also compare and contrast control and knockouts with NZB. We propose the use of NZB because our preliminary results indicates that the NZB acinar cells have significantly less amounts of NKCC1 than controls. Aim 2: Hypothesis: Apical membrane BKca channels contribute to normal fluid production of the lacrimal gland. We will test control (C57) and Bkca Beta 1 knockout mice. Aim 3: Hypothesis: PKC activity affects fluid production via regulation of K channels and/or NKCC1 transporters. We will test controls, NZB and the two knockouts (Beta1 and NKCC1).

描述（由申请人提供）：泪腺分泌的液体是眼泪的重要组成部分，估计约占角膜液体体积的50% （Walcott, 1998）。最近对小鼠泪腺产液的原位测量表明，受刺激的流量为0.2-0.6 uL/分钟（Walcott等人，2002；Paranyuk等人，2001；Moore等人，2000）。我们的初步数据表明NKCC1 Na+， K+， 2CI-共转运体和Bkca通道在泪腺的液体产生中起重要作用。我们建议使用敲除小鼠和NZB小鼠（受损的液体流动，Paranyuk等人，2001年）来研究它们各自在泪腺产生液体中的作用。此外，我们建议使用PKC的激活剂和抑制剂来描述其在调节NKCC1共转运体和Bkca通道中的作用。这两个系统都被证明受到PKC的影响/调节/调节（Standen and Quayle, 1998;Zhou etal., 2001; Clerice etal）。, 1995)。我们的建议主要针对以下假设：