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) Services Email this article to a friend Similar articles in this journal 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 Google Scholar Google Scholar Articles by Wang, L. Articles by Jacob, T. J. C. Search for Related Content PubMed PubMed Citation Articles by Wang, L. Articles by Jacob, T. J. C.

The role of ClC-3 in volume-activated chloride currents and volume regulation in bovine epithelial cells demonstrated by antisense inhibition

Liwei Wang, Lixin Chen and Tim J. C. Jacob

1. A chloride current with mild outward rectification was induced in the native bovine non-pigmented ciliary epithelial (NPCE) cells by a 23 % hypotonic solution. The current showed no or little inactivation at depolarized steps.

2. ATP blocked 88 and 61 % of the outward and inward components of the volume-activated chloride current (I_Cl,vol) with an IC₅₀ of 5·3 and 9·6 mM, respectively.

3. The volume-activated chloride current was decreased and the activation of the current was delayed by inhibiting endogenous ClC-3 expression using a ClC-3 antisense oligonucleotide. The inhibition of the current as a function of antisense concentration was asymptotic with a maximum about 60 %. The remaining current was probably not derived from ClC-3 and was inhibited by ATP.

4. ClC-3 expression in the bovine NPCE cells was verified by immunofluorescence studies. ClC-3 immunofluorescence was distributed throughout the cells but with the predominant location within the nucleus. The expression of ClC-3 protein was diminished by the ClC-3 antisense oligonucleotide with the greatest diminution occurring in the nuclear region.

5. The size of the volume-activated chloride current was positively correlated with the ClC-3 immunofluorescence level.

6. Regulatory volume decrease of the NPCE cells was reduced by ClC-3 antisense oligonucleotide.

7. We conclude that endogenous ClC-3 is associated with the volume-activated chloride current and is involved in cell volume regulation, but that it can only contribute towards a proportion of the current in NPCE cells.

8. The nuclear predominance of ClC-3 immunofluorescence in NPCE cells, the absence of basal activity of chloride current and the marked pharmacological differences between I_ClC-3 and I_Cl,vol argue against ClC-3 being the only, or even the main, volume-activated chloride channel in NPCE cells.

This article has been cited by other articles:

				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]

				L Lemonnier, R Lazarenko, Y Shuba, S Thebault, M Roudbaraki, G Lepage, N Prevarskaya, and R Skryma Alterations in the regulatory volume decrease (RVD) and swelling-activated Cl-current associated with neuroendocrine differentiation of prostate cancer epithelial cells Endocr. Relat. Cancer, June 1, 2005; 12(2): 335 - 349. [Abstract] [Full Text] [PDF]

				O. A. Candia, C.-H. To, R. M. Gerometta, and A. C. Zamudio Spontaneous Fluid Transport across Isolated Rabbit and Bovine Ciliary Body Preparations Invest. Ophthalmol. Vis. Sci., March 1, 2005; 46(3): 939 - 947. [Abstract] [Full Text] [PDF]

				C. W. Do, W. Lu, C. H. Mitchell, and M. M. Civan Inhibition of Swelling-Activated Cl- Currents by Functional Anti-ClC-3 Antibody in Native Bovine Non-Pigmented Ciliary Epithelial Cells Invest. Ophthalmol. Vis. Sci., March 1, 2005; 46(3): 948 - 955. [Abstract] [Full Text] [PDF]

				J.-G. Zhou, J.-L. Ren, Q.-y. Qiu, H. He, and Y.-Y. Guan Regulation of Intracellular Cl- Concentration through Volume-regulated ClC-3 Chloride Channels in A10 Vascular Smooth Muscle Cells J. Biol. Chem., February 25, 2005; 280(8): 7301 - 7308. [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]

				S. Yamamoto-Mizuma, G.-X. Wang, L. L. Liu, K. Schegg, W. J. Hatton, D. Duan, T. L. B. Horowitz, F. S. Lamb, and J. R. Hume Altered properties of volume-sensitive osmolyte and anion channels (VSOACs) and membrane protein expression in cardiac and smooth muscle myocytes from Clcn3-/- mice J. Physiol., June 1, 2004; 557(2): 439 - 456. [Abstract] [Full Text] [PDF]

				N. G. Jin, J. K. Kim, D. K. Yang, S. J. Cho, J. M. Kim, E. J. Koh, H. C. Jung, I. So, and K. W. Kim Fundamental role of ClC-3 in volume-sensitive Cl- channel function and cell volume regulation in AGS cells Am J Physiol Gastrointest Liver Physiol, November 1, 2003; 285(5): G938 - G948. [Abstract] [Full Text] [PDF]

				G.-X. Wang, W. J. Hatton, G. L. Wang, J. Zhong, I. Yamboliev, D. Duan, and J. R. Hume Functional effects of novel anti-ClC-3 antibodies on native volume-sensitive osmolyte and anion channels in cardiac and smooth muscle cells Am J Physiol Heart Circ Physiol, October 1, 2003; 285(4): H1453 - H1463. [Abstract] [Full Text] [PDF]

				R. Mohammad-Panah, R. Harrison, S. Dhani, C. Ackerley, L.-J. Huan, Y. Wang, and C. E. Bear The Chloride Channel ClC-4 Contributes to Endosomal Acidification and Trafficking J. Biol. Chem., August 1, 2003; 278(31): 29267 - 29277. [Abstract] [Full Text] [PDF]

				M. L. Olsen, S. Schade, S. A. Lyons, M. D. Amaral, and H. Sontheimer Expression of Voltage-Gated Chloride Channels in Human Glioma Cells J. Neurosci., July 2, 2003; 23(13): 5572 - 5582. [Abstract] [Full Text] [PDF]

				K. Juhl, M. Hoy, H. L. Olsen, K. Bokvist, A. M. Efanov, E. K. Hoffmann, and J. Gromada cPLA2{alpha}-evoked formation of arachidonic acid and lysophospholipids is required for exocytosis in mouse pancreatic {beta}-cells Am J Physiol Endocrinol Metab, July 1, 2003; 285(1): E73 - E81. [Abstract] [Full Text] [PDF]

				G.-L. Wang, X.-R. Wang, M.-J. Lin, H. He, X.-J. Lan, and Y.-Y. Guan Deficiency in ClC-3 Chloride Channels Prevents Rat Aortic Smooth Muscle Cell Proliferation Circ. Res., November 15, 2002; 91 (10): e28 - e32. [Abstract] [Full Text] [PDF]

				M. Hermoso, C. M. Satterwhite, Y. N. Andrade, J. Hidalgo, S. M. Wilson, B. Horowitz, and J. R. Hume ClC-3 Is a Fundamental Molecular Component of Volume-sensitive Outwardly Rectifying Cl- Channels and Volume Regulation in HeLa Cells and Xenopus laevis Oocytes J. Biol. Chem., October 11, 2002; 277(42): 40066 - 40074. [Abstract] [Full Text] [PDF]

				L. Chen, L. Wang, L. Zhu, S. Nie, J. Zhang, P. Zhong, B. Cai, H. Luo, and T. J. C. Jacob Cell cycle-dependent expression of volume-activated chloride currents in nasopharyngeal carcinoma cells Am J Physiol Cell Physiol, October 1, 2002; 283(4): C1313 - C1323. [Abstract] [Full Text] [PDF]

				C. Shi, S. Barnes, M. Coca-Prados, and M. E. M. Kelly Protein Tyrosine Kinase and Protein Phosphatase Signaling Pathways Regulate Volume-Sensitive Chloride Currents in a Nonpigmented Ciliary Epithelial Cell Line Invest. Ophthalmol. Vis. Sci., May 1, 2002; 43(5): 1525 - 1532. [Abstract] [Full Text] [PDF]

				T. J. Jentsch, V. Stein, F. Weinreich, and A. A. Zdebik Molecular Structure and Physiological Function of Chloride Channels Physiol Rev, April 1, 2002; 82(2): 503 - 568. [Abstract] [Full Text] [PDF]

				C. B. Ransom, J. T. O'Neal, and H. Sontheimer Volume-Activated Chloride Currents Contribute to the Resting Conductance and Invasive Migration of Human Glioma Cells J. Neurosci., October 1, 2001; 21(19): 7674 - 7683. [Abstract] [Full Text] [PDF]

				N. K. Wills and P. Fong ClC Chloride Channels in Epithelia: Recent Progress and Remaining Puzzles Physiology, August 1, 2001; 16(4): 161 - 166. [Abstract] [Full Text] [PDF]

				A. Y. Estevez, T. Bond, and K. Strange Regulation of ICl,swell in neuroblastoma cells by G protein signaling pathways Am J Physiol Cell Physiol, July 1, 2001; 281(1): C89 - C98. [Abstract] [Full Text] [PDF]

				C.-H. To, C.-W. Do, A. C. Zamudio, and O. A. Candia Model of ionic transport for bovine ciliary epithelium: effects of acetazolamide and HCO3- Am J Physiol Cell Physiol, June 1, 2001; 280(6): C1521 - C1530. [Abstract] [Full Text] [PDF]

				F. S. Lamb, R. W. Graeff, G. H. Clayton, R. L. Smith, B. C. Schutte, and P. B. McCray Jr. Ontogeny of CLCN3 Chloride Channel Gene Expression in Human Pulmonary Epithelium Am. J. Respir. Cell Mol. Biol., April 1, 2001; 24(4): 376 - 381. [Abstract] [Full Text]

				K. Nehrke, T. Begenisich, J. Pilato, and J. E. Melvin Model Organisms: New Insights: Into Ion Channel and Transporter Function. Caenorhabditis elegans ClC-type chloride channels: novel variants and functional expression Am J Physiol Cell Physiol, December 1, 2000; 279(6): C2052 - C2066. [Abstract] [Full Text] [PDF]

				X. Li, K. Shimada, L. A. Showalter, and S. A. Weinman Biophysical Properties of ClC-3 Differentiate It from Swelling-activated Chloride Channels in Chinese Hamster Ovary-K1 Cells J. Biol. Chem., November 10, 2000; 275(46): 35994 - 35998. [Abstract] [Full Text] [PDF]

				R. Mohammad-Panah, K. Gyomorey, J. Rommens, M. Choudhury, C. Li, Y. Wang, and C. E. Bear ClC-2 Contributes to Native Chloride Secretion by a Human Intestinal Cell Line, Caco-2 J. Biol. Chem., March 9, 2001; 276(11): 8306 - 8313. [Abstract] [Full Text] [PDF]

				K.-H. Weylandt, M. A. Valverde, M. Nobles, S. Raguz, J. S. Amey, M. Diaz, C. Nastrucci, C. F. Higgins, and A. Sardini Human ClC-3 Is Not the Swelling-activated Chloride Channel Involved in Cell Volume Regulation J. Biol. Chem., May 11, 2001; 276(20): 17461 - 17467. [Abstract] [Full Text] [PDF]

				M. Christensen and K. Strange Developmental Regulation of a Novel Outwardly Rectifying Mechanosensitive Anion Channel in Caenorhabditis elegans J. Biol. Chem., November 21, 2001; 276(48): 45024 - 45030. [Abstract] [Full Text] [PDF]