Vasoactive intestinal peptide, forskolin, and genistein increase apical CFTR trafficking in the rectal gland of the spiny dogfish, Squalus acanthias - Acute regulation of CFTR trafficking in an intact epithelium

被引:78
作者
Lehrich, RW
Aller, SG
Webster, P
Marino, CR
Forrest, JN
机构
[1] Yale Univ, Sch Med, Dept Med, New Haven, CT 06510 USA
[2] Yale Univ, Sch Med, Dept Cell Biol, New Haven, CT 06510 USA
[3] Humboldt Univ, Klinikum Rudolf Virchow, Franz Volhard Clin, Max Delbruck Ctr Mol Med, D-13122 Berlin, Germany
[4] Univ Tennessee, Dept Med, Memphis, TN 38104 USA
[5] Univ Tennessee, Dept Physiol & Biophys, Memphis, TN 38104 USA
[6] Mt Desert Isl Biol Lab, Salisbury Cove, ME 04672 USA
关键词
cystic fibrosis transmembrane conductance regulator; CFTR; shark rectal gland; membrane trafficking; forskolin; genistein;
D O I
10.1172/JCI803
中图分类号
R-3 [医学研究方法]; R3 [基础医学];
学科分类号
1001 ;
摘要
Defective trafficking of the cystic fibrosis transmembrane conductance regulator (CFTR) is the most common cause of cystic fibrosis. In chloride-secreting epithelia, it is well established that CFTR localizes to intracellular organelles and to apical membranes, However, it is controversial whether secretagogues regulate the trafficking of CFTR. To investigate whether acute hormonal stimulation of chloride secretion is coupled to the trafficking of CFTR, we used the intact shark rectal gland, a model tissue in which salt secretion is dynamically regulated and both chloride secretion and cellular CFTR immunofluorescence can be quantified in parallel. In rectal glands perfused under basal conditions without secretagogues, Cl- secretion was 151+/-65 mu eq/h/g. Vasoactive intestinal peptide (VIP), forskolin, and genistein led to 10-, 6-, and 4-fold increases in Cl- secretion. In basal glands, quantitative confocal microscopy revealed CFTR immunofluorescence extending from the apical membrane deeply into the cell (7.28+/-0.35 mu m). During stimulation with secretagogues, apical extension of CFTR immunofluorescence into the cell was reduced significantly to 3.24+/-0.08 mu m by VIP, 4.08+/-0.13 by forskolin, and 3.19+/-0.1 by genistein (P < 0.001). Moreover, the peak intensity of CFTR fluorescence shifted towards the apical membrane (peak fluorescence 2.5+/-0.13 mu m basal vs, 1.51+/-0.06, 1.77+/-0.1, and 1.38+/-0.05 for VIP, forskolin, and genistein; all P < 0.001). The increase in both Cl- secretion and apical CFTR trafficking reversed to basal values after removal of VIP, These data provide the first quantitative morphological evidence for acute hormonal regulation of CFTR trafficking in an intact epithelial tissue.
引用
收藏
页码:737 / 745
页数:9
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