Ko Shigeru B H, Naruse Satoru, Kitagawa Motoji, Ishiguro Hiroshi, Furuya Sonoko, Mizuno Nobumasa, Wang Youxue, Yoshikawa Toshiyuki, Suzuki Atsushi, Shimano Shoko, Hayakawa Tetsuo
Internal Medicine II, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan.
Am J Physiol Gastrointest Liver Physiol. 2002 Feb;282(2):G324-31. doi: 10.1152/ajpgi.00198.2001.
The aquaporin (AQP) family of water channels is distributed ubiquitously in many epithelia and plays a fundamental role in transmembrane water transport. The aim of this study is to identify the water transport pathway in pancreatic duct cells where most of the HCO-rich fluid originates. Using digital videomicroscopy, we measured the osmotic water permeability (P(f)) of pancreatic duct epithelium by exposing isolated rat interlobular ducts to the hypotonic solution (145 mosM). To identify mRNA and protein of AQPs expressed in duct cells, we conducted RT-PCR analysis and immunohistochemistry of the isolated duct and pancreas. The calculated P(f) (160-230 microm/s) of the isolated ducts was significantly reduced to 16-35 microm/s by 80-90% with either basolateral or luminal applications of HgCl(2). Fluid secretion evoked by secretin was almost completely abolished by a basolateral or luminal application of HgCl(2). A large amount of AQP1 and small amounts of AQP5 transcripts were detected in the isolated duct cells by RT-PCR. AQP1, but not AQP5, immunoreactivity was present in both luminal and basolateral membranes of the interlobular duct cells. Mercury-sensitive water channels are present in both luminal and basolateral membranes of rat pancreatic ducts. AQP1 of the known AQPs appears to be the main water pathway in interlobular ducts.
水通道蛋白(AQP)家族的水通道广泛分布于多种上皮组织中,在跨膜水转运中发挥着重要作用。本研究的目的是确定富含碳酸氢根的液体大部分产生的胰腺导管细胞中的水转运途径。我们使用数字视频显微镜,通过将分离的大鼠小叶间导管暴露于低渗溶液(145 mosM)中来测量胰腺导管上皮的渗透水通透性(P(f))。为了鉴定导管细胞中表达的水通道蛋白的mRNA和蛋白质,我们对分离的导管和胰腺进行了逆转录聚合酶链反应(RT-PCR)分析和免疫组织化学研究。分离导管的计算P(f)(160 - 230微米/秒)在基底外侧或管腔应用氯化汞后显著降低80 - 90%,降至16 - 35微米/秒。促胰液素引起的液体分泌在基底外侧或管腔应用氯化汞后几乎完全被消除。通过RT-PCR在分离的导管细胞中检测到大量的水通道蛋白1(AQP1)转录本和少量的水通道蛋白5(AQP5)转录本。小叶间导管细胞的管腔和基底外侧膜中均存在AQP1免疫反应性,但不存在AQP5免疫反应性。大鼠胰腺导管的管腔和基底外侧膜中均存在汞敏感的水通道。已知水通道蛋白中的AQP1似乎是小叶间导管中的主要水转运途径。