Linked Life Data

18077605

Source:http://linkedlifedata.com/resource/pubmed/id/18077605

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2008-2-15
pubmed:abstractText
To determine the effects of chloride channel 3 (ClC-3) knockdown and overexpression on lysophosphatidic acid (LPA)- and volume-regulated anion channel Cl(-) currents (I(Cl,LPA) and I(Cl,VRAC), respectively), cell differentiation, and cell volume regulation, a short hairpin RNA (shRNA) expression system based on a mouse U6 promoter was used to knock down ClC-3 in human corneal keratocytes and human fetal lung fibroblasts. ClC-3 overexpression was achieved by electroporating full-length ClC-3, within a pcDNA3.1 vector, into these two cell lines. RT-PCR and Western blot analysis were used to detect ClC-3 mRNA and protein levels. Whole cell perforated patch-clamp recording was used to measure I(Cl,LPA) and I(Cl,VRAC) currents, and fluorescence-activated cell sorting analysis was used to measure cell volume regulation. ClC-3 knockdown significantly decreased I(Cl,LPA) and I(Cl,VRAC) activity in the presence of transforming growth factor-beta(1) (TGF-beta(1)) compared with controls, whereas ClC-3 overexpression resulted in increased I(Cl,LPA) activity in the absence of TGF-beta(1). ClC-3 knockdown also resulted in a reduction of alpha-smooth muscle actin (alpha-SMA) protein levels in the presence of TGF-beta(1), whereas ClC-3 overexpression increased alpha-SMA protein expression in the absence of TGF-beta(1). In addition, keratocytes transfected with ClC-3 shRNA had a significantly blunted regulatory volume decrease response following hyposmotic stimulation compared with controls. These data confirm that ClC-3 is important in VRAC function and cell volume regulation, is associated with the I(Cl,LPA) current activity, and participates in the fibroblast-to-myofibroblast transition.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0363-6143
pubmed:author
pubmed:issnType
Print
pubmed:volume
294
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
C535-42
pubmed:dateRevised
2011-7-15
pubmed:meshHeading
pubmed-meshheading:18077605-Actins, pubmed-meshheading:18077605-Cell Differentiation, pubmed-meshheading:18077605-Cell Line, pubmed-meshheading:18077605-Cell Size, pubmed-meshheading:18077605-Chloride Channels, pubmed-meshheading:18077605-Chlorides, pubmed-meshheading:18077605-Down-Regulation, pubmed-meshheading:18077605-Fibroblasts, pubmed-meshheading:18077605-Fibrosis, pubmed-meshheading:18077605-Flow Cytometry, pubmed-meshheading:18077605-Humans, pubmed-meshheading:18077605-Keratinocytes, pubmed-meshheading:18077605-Lysophospholipids, pubmed-meshheading:18077605-Membrane Potentials, pubmed-meshheading:18077605-Myocytes, Smooth Muscle, pubmed-meshheading:18077605-Patch-Clamp Techniques, pubmed-meshheading:18077605-RNA Interference, pubmed-meshheading:18077605-Transforming Growth Factor beta1, pubmed-meshheading:18077605-Wound Healing
pubmed:year
2008
pubmed:articleTitle
ClC-3 is required for LPA-activated Cl- current activity and fibroblast-to-myofibroblast differentiation.
pubmed:affiliation
Department of Physiology, University of Tennessee Health Science Center, 894 Union Ave., Memphis, TN 38163, USA.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't