SHERP

Protease-activated receptor regulation of Cl– secretion in Calu-3 cells requires prostaglandin release and CFTR activation

Cited 0 time in webofscience Cited 37 time in scopus
Authors
Palmer, Melissa L.; Lee, So Yeong; Maniak, Peter J.; Carlson, Dan; Fahrenkrug, Scott C.; O’Grady, Scott M.
Issue Date
2006
Publisher
The American Physiological Society
Citation
Am J Physiol Cell Physiol 290: C1189–C1198, 2006
Keywords
cystic fibrosis transmembrane conductance regulatorKCNQ1calciumactivated potassium channelsKCNN4cAMP
Abstract
Human lung epithelial (Calu-3) cells were used to investigate the effects of protease-activated receptor (PAR) stimulation on Cl– secretion. Quantitative RT-PCR (QRT-PCR) showed that Calu-3 cells express PAR-1, -2, and -3 receptor mRNAs, with PAR-2 mRNA in greatest abundance. Addition of either thrombin or the PAR-2 agonist peptide SLIGRL to the basolateral solution of monolayers mounted in Ussing chambers produced a rapid increase in short-circuit current (Isc: thrombin, 21 ± 2 µA; SLIGRL, 83 ± 22 µA), which returned to baseline within 5 min after stimulation. Pretreatment of monolayers with the cell-permeant Ca2+-chelating agent BAPTA-AM (50 µM) abolished the increase in Isc produced by SLIGRL. When monolayers were treated with the cyclooxygenase inhibitor indomethacin (10 µM), nearly complete inhibition of both the thrombin- and SLIGRL-stimulated Isc was observed. In addition, basolateral treatment with the PGE2 receptor antagonist AH-6809 (25 µM) significantly inhibited the effects of SLIGRL on Isc. QRT-PCR revealed that Calu-3 cells express mRNAs for CFTR, the Ca2+-activated KCNN4 K+ channel, and the KCNQ1 K+ channel subunit, which, in association with KCNE3, is known to be regulated by cAMP. Stimulation with SLIGRL produced an increase in apical Cl– conductance that was blocked in cells expressing short hairpin RNAs designed to target CFTR. These results support the conclusion that PAR stimulation of Cl– secretion occurs by an indirect mechanism involving the synthesis and release of prostaglandins. In addition, PAR-stimulated Cl– secretion requires activation of CFTR and at least two distinct K+ channels located in the basolateral membrane.
ISSN
0363-6143 (print)
1522-1563 (online)
Language
English
URI
http://hdl.handle.net/10371/6550
DOI
https://doi.org/10.1152/ajpcell.00464.2005
Files in This Item:
There are no files associated with this item.
Appears in Collections:
College of Veterinary Medicine (수의과대학)Dept. of Veterinary Medicine (수의학과)Journal Papers (저널논문_수의학과)
  • mendeley

Items in S-Space are protected by copyright, with all rights reserved, unless otherwise indicated.

Browse