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

J. Biol. Chem., Vol. 281, Issue 22, 15572-15581, June 2, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/22/15572    most recent M600090200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wang, Y. Articles by Meissner, G. Search for Related Content PubMed PubMed Citation Articles by Wang, Y. Articles by Meissner, G. Social Bookmarking                      What's this?

Knocking Down Type 2 but Not Type 1 Calsequestrin Reduces Calcium Sequestration and Release in C₂C₁₂ Skeletal Muscle Myotubes^*

Ying Wang, Le Xu, Hongzhe Duan, Daniel A. Pasek, Jerry P. Eu^¶, and Gerhard Meissner¹

From the Departments of Biochemistry and Biophysics, and Cell and Molecular Physiology, Gene Therapy Center, University of North Carolina, Chapel Hill, North Carolina 27599-7260 and the ^¶Division of Pulmonary, Allergy and Critical Care Medicine, Duke University Medical Center, Durham, North Carolina 27710

We examined the roles of type 1 and type 2 calsequestrins (CSQ1 and CSQ2) in stored Ca²⁺ release of C₂C₁₂ skeletal muscle myotubes. Transduction of C₂C₁₂ myoblasts with CSQ1 or CSQ2 small interfering RNAs effectively reduced the expression of targeted CSQ protein to near undetectable levels. As compared with control infected or CSQ1 knockdown myotubes, CSQ2 and CSQ1/CSQ2 knockdown myotubes had significantly reduced stored Ca²⁺ release evoked by activators of intracellular Ca²⁺ release channel/ryanodine receptor (10 mM caffeine, 200 µM 4-chloro-m-cresol, or 10 mM KCl). Thus, CSQ1 is not essential for effective stored Ca²⁺ release in C₂C₁₂ myotubes despite our in vitro studies suggesting that CSQ1 may enhance ryanodine receptor channel activity. To determine the basis of the reduced stored Ca²⁺ release in CSQ2 knockdown myotubes, we performed immunoblot analyses and found a significant reduction in both sarco/endoplasmic reticulum Ca²⁺-ATPase and skeletal muscle ryanodine receptor proteins in CSQ2 and CSQ1/CSQ2 knockdown myotubes. Moreover, these knockdown myotubes exhibited reduced Ca²⁺ uptake and reduced stored Ca²⁺ release by UTP (400 µM) that activates a different family of intracellular Ca²⁺ release channels (inositol 1,4,5-trisphosphate receptors). Taken together, our data suggest that knocking down CSQ2, but not CSQ1, leads to reduced Ca²⁺ storage and release in C₂C₁₂ myotubes.

Received for publication, January 4, 2006 , and in revised form, March 14, 2006.

^* This work was supported by National Institutes of Health Grants HL 73051 and AR18687. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed. Tel.: 919-966-5021; Fax: 919-966-2852; E-mail: meissner{at}med.unc.edu.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				G. Meissner, Y. Wang, L. Xu, and J. P. Eu Silencing genes of sarcoplasmic reticulum proteins clarifies their roles in excitation\#8211;contraction coupling J. Physiol., July 1, 2009; 587(13): 3089 - 3090. [Full Text] [PDF]

				L. Royer and E. Ríos Deconstructing calsequestrin. Complex buffering in the calcium store of skeletal muscle J. Physiol., July 1, 2009; 587(13): 3101 - 3111. [Abstract] [Full Text] [PDF]

				F. Protasi, C. Paolini, and M. Dainese Calsequestrin-1: a new candidate gene for malignant hyperthermia and exertional/environmental heat stroke J. Physiol., July 1, 2009; 587(13): 3095 - 3100. [Abstract] [Full Text] [PDF]

				M. Dainese, M. Quarta, A. D. Lyfenko, C. Paolini, M. Canato, C. Reggiani, R. T. Dirksen, and F. Protasi Anesthetic- and heat-induced sudden death in calsequestrin-1-knockout mice FASEB J, June 1, 2009; 23(6): 1710 - 1720. [Abstract] [Full Text] [PDF]

				R. M. Murphy, N. T. Larkins, J. P. Mollica, N. A. Beard, and G. D. Lamb Calsequestrin content and SERCA determine normal and maximal Ca2+ storage levels in sarcoplasmic reticulum of fast- and slow-twitch fibres of rat J. Physiol., January 15, 2009; 587(2): 443 - 460. [Abstract] [Full Text] [PDF]

				L. Xu, Y. Wang, N. Yamaguchi, D. A. Pasek, and G. Meissner Single Channel Properties of Heterotetrameric Mutant RyR1 Ion Channels Linked to Core Myopathies J. Biol. Chem., March 7, 2008; 283(10): 6321 - 6329. [Abstract] [Full Text] [PDF]

				C. Paolini, M. Quarta, A. Nori, S. Boncompagni, M. Canato, P. Volpe, P. D. Allen, C. Reggiani, and F. Protasi Reorganized stores and impaired calcium handling in skeletal muscle of mice lacking calsequestrin-1 J. Physiol., September 1, 2007; 583(2): 767 - 784. [Abstract] [Full Text] [PDF]

				J. H. Cho, K. M. Ko, G. Singaruvelu, W. Lee, G. B. Kang, S.-H. Rho, B.-J. Park, J.-R. Yu, H. Kagawa, S. H. Eom, et al. Functional importance of polymerization and localization of calsequestrin in C. elegans J. Cell Sci., May 1, 2007; 120(9): 1551 - 1558. [Abstract] [Full Text] [PDF]

				S. Pouvreau, L. Royer, J. Yi, G. Brum, G. Meissner, E. Rios, and J. Zhou Ca2+ sparks operated by membrane depolarization require isoform 3 ryanodine receptor channels in skeletal muscle PNAS, March 20, 2007; 104(12): 5235 - 5240. [Abstract] [Full Text] [PDF]