Water Transport in the Lacrimal Gland

描述（由申请人提供）：泪腺分泌的液体是泪液的基本成分，估计约占角膜液体量的50%（Walcott，1998）。最近，小鼠泪腺的液体产生的原位测量已经证明了0.2-0.6 μ L/分钟的刺激流速（Walcott等人，2002; Paranyuk等人，2001;摩尔等人，2000）。我们的初步数据表明，NKCC 1 Na+，K+，2CI-协同转运蛋白和Bkca通道在泪腺的液体产生中起重要作用。我们建议使用基因敲除小鼠沿着NZB小鼠（受损的流体流动，Paranyuk等人，2001）以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NKCC 1协同转运蛋白和Bkca通道调节中的作用。两种系统均已显示受PKC影响/调节/调节（Standen和Quayle，1998;Zhou等人，2001年; Clerice埃塔尔，1995年）。我们的建议侧重于解决以下假设： 目标1假设：盐共转运蛋白（NKCC 1）的基底外侧阻断将显著降低对照中的刺激液流量，但与NKCC 1敲除流速相似。我们还将比较和对比控制和淘汰赛与NZB。我们建议使用NZB，因为我们的初步结果表明，NZB腺泡细胞的NKCC 1的量显着低于对照组。 目的2：假设：泪腺顶膜BKca通道有助于泪腺的正常液体产生。我们将测试对照（C57）和Bkca β 1敲除小鼠。 目的3：假设：PKC活性通过调节K通道和/或NKCC 1转运蛋白影响液体产生。我们将测试控制，NZB和两个敲除（Beta1和NKCC 1）。

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).