HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: 285/4/H1453    most recent 00244.2003v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (35) Citing Articles via Google Scholar Google Scholar Articles by Wang, G.-X. Articles by Hume, J. R. Search for Related Content PubMed PubMed Citation Articles by Wang, G.-X. Articles by Hume, J. R.

Functional effects of novel anti-ClC-3 antibodies on native volume-sensitive osmolyte and anion channels in cardiac and smooth muscle cells

Center of Biomedical Research Excellence, Department of Pharmacology, University of Nevada School of Medicine Reno, Nevada 89557-0046

Submitted 18 March 2003 ; accepted in final form 12 June 2003

Whether ClC-3 encodes volume-sensitive organic osmolyte and anion channels (VSOACs) remains controversial. We have shown previously that native VSOACs in some cardiac and vascular myocytes were blocked by a commercial anti-ClC-3 carboxy terminal antibody (Alm C_592–661 antibody), although recent studies have raised questions related to the specificity of Alm C_592–661 antibody. Therefore, we have developed three new anti-ClC-3 antibodies and investigated their functional effects on native VSOACs in freshly isolated canine pulmonary artery smooth muscle cells (PASMCs) and guinea pig cardiac myocytes. These new antibodies produced a common prominent immunoreactive band with an apparent molecular mass of 90–92 kDa in the guinea pig heart and PASMCs, and a similar molecular mass immunoreactive band was observed in the brain from homozgygous Clcn3^+/+ mice but not from homozygous Clcn3^–/– mice. VSOACs elicited by hypotonic cell swelling in PASMCs and guinea pig atrial myocytes were nearly completely abolished by intracellular dialysis with two new anti-ClC-3 antibodies specifically targeting the ClC-3 carboxy (C_670–687 antibody) and amino terminus (A_1–14 antibody). This inhibition of native VSOACs can be attributed to a specific interaction with endogenous ClC-3, because 1) preabsorption of the antibodies with corresponding antigens prevented the inhibitory effects, 2) extracellular application of a new antibody raised against an extracellular epitope (Ex_133–148) of ClC-3 failed to inhibit native VSOACs in PASMCs, 3) intracellular dialysis with an antibody targeting Kv1.1 potassium channels failed to inhibit native VSOACs in guinea pig atrial myocytes, and 4) anti-ClC-3 C_670–687 antibody had no effects on swelling-induced augmentation of the slow component of the delayed rectifying potassium current in guinea pig ventricular myocytes, although VSOACs in the same cells were inhibited by the antibody. These results confirm that endogenous ClC-3 is an essential molecular entity responsible for native VSOACs in PASMCs and guinea pig cardiac myocytes.

chloride channels; cell volume

This article has been cited by other articles:

				D. Duan Phenomics of cardiac chloride channels: the systematic study of chloride channel function in the heart J. Physiol., May 1, 2009; 587(10): 2163 - 2177. [Abstract] [Full Text] [PDF]

				Z. Yin, Y. Tong, H. Zhu, and M. A. Watsky ClC-3 is required for LPA-activated Cl- current activity and fibroblast-to-myofibroblast differentiation Am J Physiol Cell Physiol, February 1, 2008; 294(2): C535 - C542. [Abstract] [Full Text] [PDF]

				C.H. Yeung, J.P. Barfield, and T.G. Cooper Chloride Channels in Physiological Volume Regulation of Human Spermatozoa Biol Reprod, November 1, 2005; 73(5): 1057 - 1063. [Abstract] [Full Text] [PDF]

				N. J. Ernest, A. K. Weaver, L. B. Van Duyn, and H. W. Sontheimer Relative contribution of chloride channels and transporters to regulatory volume decrease in human glioma cells Am J Physiol Cell Physiol, June 1, 2005; 288(6): C1451 - C1460. [Abstract] [Full Text] [PDF]

				G.-X. Wang, C. McCrudden, Y.-P. Dai, B. Horowitz, J. R. Hume, and I. A. Yamboliev Hypotonic activation of volume-sensitive outwardly rectifying chloride channels in cultured PASMCs is modulated by SGK Am J Physiol Heart Circ Physiol, August 1, 2004; 287(2): H533 - H544. [Abstract] [Full Text] [PDF]

				L. Lemonnier, Y. Shuba, A. Crepin, M. Roudbaraki, C. Slomianny, B. Mauroy, B. Nilius, N. Prevarskaya, and R. Skryma Bcl-2-Dependent Modulation of Swelling-Activated Cl- Current and ClC-3 Expression in Human Prostate Cancer Epithelial Cells Cancer Res., July 15, 2004; 64(14): 4841 - 4848. [Abstract] [Full Text] [PDF]