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Research Article Free access | 10.1172/JCI803

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.

R W Lehrich, S G Aller, P Webster, C R Marino, and J N Forrest Jr

Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

Find articles by Lehrich, R. in: PubMed | Google Scholar

Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

Find articles by Aller, S. in: PubMed | Google Scholar

Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

Find articles by Webster, P. in: PubMed | Google Scholar

Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

Find articles by Marino, C. in: PubMed | Google Scholar

Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

Find articles by Forrest, J. in: PubMed | Google Scholar

Published February 15, 1998 - More info

Published in Volume 101, Issue 4 on February 15, 1998
J Clin Invest. 1998;101(4):737–745. https://doi.org/10.1172/JCI803.
© 1998 The American Society for Clinical Investigation
Published February 15, 1998 - Version history
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Abstract

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 microeq/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 micron). During stimulation with secretagogues, apical extension of CFTR immunofluorescence into the cell was reduced significantly to 3.24+/-0.08 micron 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 micron 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.

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