SPI-0211 Activates T84 Cell Cl- Transport and Recombinant Human ClC-2 Cl- Currents
نویسندگان
چکیده
The purpose of this study was to determine the mechanism of action of SPI-0211 (Lubiprostone), a novel bicyclic fatty acid, under development for the treatment of bowel dysfunction. Adult rabbit intestine was shown to contain mRNA for ClC-2 by RT-PCR, Northern 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 EC 50 of ≈ 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 a human epithelial kidney cell line (HEK-293) 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
منابع مشابه
TRANSLATIONAL PHYSIOLOGY SPI-0211 activates T84 cell chloride transport and recombinant human ClC-2 chloride currents
Cuppoletti, John, Danuta H. Malinowska, Kirti P. Tewari, Qiu-ju Li, Ann M. Sherry, Myra L. Patchen, and Ryuji Ueno. SPI-0211 activates T84 cell chloride transport and recombinant human ClC-2 chloride currents. Am J Physiol Cell Physiol 287: C1173– C1183, 2004. First published June 22, 2004; doi:10.1152/ajpcell. 00528.2003.—The purpose of this study was to determine the mechanism of action of SP...
متن کاملSPI-0211 activates T84 cell chloride transport and recombinant human ClC-2 chloride currents.
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...
متن کاملDifferentiation between human ClC-2 and CFTR Cl- channels with pharmacological agents.
It has been difficult to separate/identify the roles of ClC-2 and CFTR in Cl(-) transport studies. Using pharmacological agents, we aimed to differentiate functionally between ClC-2 and CFTR Cl(-) channel currents. Effects of CFTR inhibitor 172 (CFTRinh172), N-(4-methylphenylsulfonyl)-N'-(4-trifluoromethylphenyl)urea (DASU-02), and methadone were examined by whole cell patch clamp on Cl(-) curr...
متن کاملA synthetic prostone activates apical chloride channels in A6 epithelial cells.
The bicyclic fatty acid lubiprostone (formerly known as SPI-0211) activates two types of anion channels in A6 cells. Both channel types are rarely, if ever, observed in untreated cells. The first channel type was activated at low concentrations of lubiprostone (<100 nM) in >80% of cell-attached patches and had a unit conductance of approximately 3-4 pS. The second channel type required higher c...
متن کاملA single point mutation reveals gating of the human ClC-5 Cl−/H+ antiporter
ClC-5 is a 2Cl(-)/1H(+) antiporter highly expressed in endosomes of proximal tubule cells. It is essential for endocytosis and mutations in ClC-5 cause Dent's disease, potentially leading to renal failure. However, the physiological role of ClC-5 is still unclear. One of the main issues is whether the strong rectification of ClC-5 currents observed in heterologous systems, with currents elicite...
متن کامل