Document Detail

ClC-3 is a fundamental molecular component of volume-sensitive outwardly rectifying Cl- channels and volume regulation in HeLa cells and Xenopus laevis oocytes.
MedLine Citation:
PMID:  12183454     Owner:  NLM     Status:  MEDLINE    
Volume-sensitive osmolyte and anion channels (VSOACs) are activated upon cell swelling in most vertebrate cells. Native VSOACs are believed to be a major pathway for regulatory volume decrease (RVD) through efflux of chloride and organic osmolytes. ClC-3 has been proposed to encode native VSOACs in Xenopus laevis oocytes and in some mammalian cells, including cardiac and vascular smooth muscle cells. The relationship between the ClC-3 chloride channel, the native volume-sensitive osmolyte and anion channel (VSOAC) currents, and cell volume regulation in HeLa cells and X. laevis oocytes was investigated using ClC-3 antisense. In situ hybridization in HeLa cells, semiquantitative and real-time PCR, and immunoblot studies in HeLa cells and X. laevis oocytes demonstrated the presence of ClC-3 mRNA and protein, respectively. Exposing both cell types to hypotonic solutions induced cell swelling and activated native VSOACs. Transient transfection of HeLa cells with ClC-3 antisense oligonucleotide or X. laevis oocytes injected with antisense cRNA abolished the native ClC-3 mRNA transcript and protein and significantly reduced the density of native VSOACs activated by hypotonically induced cell swelling. In addition, antisense against native ClC-3 significantly impaired the ability of HeLa cells and X. laevis oocytes to regulate their volume. These results suggest that ClC-3 is an important molecular component underlying VSOACs and the RVD process in HeLa cells and X. laevis oocytes.
Marcela Hermoso; Christina M Satterwhite; Yanire Naty Andrade; Jorge Hidalgo; Sean M Wilson; Burton Horowitz; Joseph R Hume
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.     Date:  2002-08-14
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  277     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  2002 Oct 
Date Detail:
Created Date:  2002-10-15     Completed Date:  2002-12-19     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  40066-74     Citation Subset:  IM    
Instituto de Ciencias Biomédicas, Facultad de Medicina Universidad de Chile, Santiago 6530499, Chile.
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MeSH Terms
Chloride Channels / chemistry*,  physiology*
Chlorides / metabolism
DNA Primers / chemistry
Hela Cells
In Situ Hybridization
Mutagenesis, Site-Directed
Oligonucleotides, Antisense / metabolism,  pharmacology
Oocytes / metabolism
Patch-Clamp Techniques
Protein Binding
RNA, Complementary / metabolism
RNA, Messenger / metabolism
Reverse Transcriptase Polymerase Chain Reaction
Time Factors
Xenopus laevis / embryology*
Grant Support
Reg. No./Substance:
0/Anions; 0/Chloride Channels; 0/Chlorides; 0/ClC-3 channel; 0/DNA Primers; 0/Oligonucleotides, Antisense; 0/RNA, Complementary; 0/RNA, Messenger

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