Channelpedia

PubMed 15213059


Referenced in Channelpedia wiki pages of: none

Automatically associated channels: ClvC2 , ClvC4



Title: SPI-0211 activates T84 cell chloride transport and recombinant human ClC-2 chloride currents.

Authors: John Cuppoletti, Danuta H Malinowska, Kirti P Tewari, Qiu-Ju Li, Ann M Sherry, Myra L Patchen, Ryuji Ueno

Journal, date & volume: Am. J. Physiol., Cell Physiol., 2004 Nov , 287, C1173-83

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/15213059


Abstract
The purpose of this study was to determine the mechanism of action of SPI-0211 (lubiprostone), a novel bicyclic fatty acid in development for the treatment of bowel dysfunction. Adult rabbit intestine was shown to contain mRNA for ClC-2 using RT-PCR, Northern blot analysis, and in situ hybridization. T84 cells grown to confluence on permeable supports were shown to express ClC-2 channel protein in the apical membrane. SPI-0211 increased electrogenic Cl- transport across the apical membrane of T84 cells, with an EC50 of approximately 18 nM measured by short-circuit current (Isc) after permeabilization of the basolateral membrane with nystatin. SPI-0211 effects on Cl- currents were also measured by whole cell patch clamp using the human embryonic kidney (HEK)-293 cell line stably transfected with either recombinant human ClC-2 or recombinant human cystic fibrosis transmembrane regulator (CFTR). In these studies, SPI-0211 activated ClC-2 Cl- currents in a concentration-dependent manner, with an EC50 of approximately 17 nM, and had no effect in nontransfected HEK-293 cells. In contrast, SPI-0211 had no effect on CFTR Cl- channel currents measured in CFTR-transfected HEK-293 cells. Activation of ClC-2 by SPI-0211 was independent of PKA. Together, these studies demonstrate that SPI-0211 is a potent activator of ClC-2 Cl- channels and suggest a physiologically relevant role for ClC-2 Cl- channels in intestinal Cl- transport after SPI-0211 administration.