CFTR-Mediated Cl- Transport in the Acinar and Duct Cells of Rabbit Lacrimal Gland
CFTR-Mediated Cl- Transport in the Acinar and Duct Cells of Rabbit Lacrimal Gland
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DOI：
10.3109/02713683.2012.675613
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发表时间：
2012-08-01
影响因子：
2
通讯作者：
Ding, Chuanqing
中科院分区：
文献类型：
--
作者：
Lu, Michael;Ding, Chuanqing
目的：我们研究了囊性纤维化跨膜传导调节因子（CFTR）在兔泪腺（LG）腺泡和导管上皮细胞Cl-转运中的作用。原代培养的LG腺泡细胞在高[Cl-]和低[Cl-]灌注液中进行全细胞膜片钳电生理记录，10 μ M毛喉素和100 μ M 3-异丁基-1-甲基黄嘌呤（IBMX）、非特异性Cl-通道阻断剂4，4 '-二硫氰基芪-2，2'磺酸（DIDS; 100 μ M）和CFTR特异性阻断剂CFTRinh-172（10 μ M）。用多光子共聚焦激光扫描显微镜对新鲜分离的LG导管进行离体活细胞[Cl-]变化成像，使用Cl-敏感的荧光染料N-[ethoxycarbonylmethyl]-6-methoxy-quinoliniumbromide.Results：全细胞膜片钳研究证实在分离的腺泡细胞中存在Cl-电流，并揭示该Cl-电流由CFTR通道介导。活细胞成像也显示存在CFTR介导的Cl-运输跨质膜的导管cells.Conclusions：我们以前的数据显示CFTR的存在下，在所有的腺泡和导管细胞内的兔LG，表达最突出的导管细胞的顶膜。目前的研究表明，CFTR是积极参与Cl-运输腺泡细胞和上皮细胞从导管段，这表明CFTR可能发挥重要作用，LG分泌。
Purpose: We investigated the role that the cystic fibrosis transmembrane conductance regulator (CFTR) may play in Cl- transport in the acinar and ductal epithelial cells of rabbit lacrimal gland (LG).Methods: Primary cultured LG acinar cells were processed for whole-cell patch-clamp electrophysiological recording of Cl- currents by using perfusion media with high and low [Cl-], 10 mu M forskolin and 100 mu M 3-isobutyl-1-methylxanthine (IBMX), the non-specific Cl- channel blocker 4,4'-disothiocyanostilbene-2, 2' sulphonic acid (DIDS; 100 mu M) and CFTRinh-172 (10 mu M), a specific blocker for CFTR. Ex vivo live cell imaging of [Cl-] changes in duct cells was performed on freshly dissected LG duct with a multiphoton confocal laser scanning microscope using a Cl- sensitive fluorescence dye, N-[ethoxycarbonylmethyl]-6-methoxy-quinolinium bromide.Results: Whole-cell patch-clamp studies demonstrated the presence of Cl- current in isolated acinar cells and revealed that this Cl- current was mediated by CFTR channel. Live cell imaging also showed the presence of CFTR-mediated Cl- transport across the plasma membrane of duct cells.Conclusions: Our previous data showed the presence of CFTR in all acinar and duct cells within the rabbit LG, with expression most prominent in the apical membranes of duct cells. The present study demonstrates that CFTR is actively involved in Cl- transport in both acinar cells and epithelial cells from duct segments, suggesting that CFTR may play a significant role in LG secretion.