QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

First published online 8 April 2003
doi: 10.1242/jcs.00447

This Article Figures Only Full Text Full Text (PDF) All Versions of this Article: jcs.00447v1 116/11/2187    most recent Alert me when this article is cited Alert me if a correction is posted 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 Jenniskens, G. J. Articles by van Kuppevelt, T. H. Search for Related Content PubMed PubMed Citation Articles by Jenniskens, G. J. Articles by van Kuppevelt, T. H.

Disturbed Ca²⁺ kinetics in N-deacetylase/N-sulfotransferase-1 defective myotubes

Guido J. Jenniskens^1,*, Maria Ringvall², Werner J. H. Koopman³, Johan Ledin², Lena Kjellén², Peter H. G. M. Willems³, Erik Forsberg², Jacques H. Veerkamp¹ and Toin H. van Kuppevelt^1,

¹ Department of Biochemistry 194, University Medical Center, NCMLS, 6500 HB Nijmegen, The Netherlands
² Department of Medical Biochemistry and Microbiology, Uppsala University, S-751 23 Uppsala, Sweden
³ Department of Biochemistry 160/Microscopical Imaging Center, University Medical Center, NCMLS, 6500 HB Nijmegen, The Netherlands
^* Present address: Biological Engineering Division, Massachusetts Institute of Technology, Cambridge, MA 02139, USA

Author for correspondence (e-mail: a.vankuppevelt{at}ncmls.kun.nl)

Accepted 28 February 2003

The biosynthesis of heparan sulfate, present on the cell surface and in the basal lamina surrounding cells, is a multistep process in which each step is mediated by a specific enzyme. The initial modification of the precursor polysaccharide, N-deacetylation followed by N-sulfation of selected N-acetyl-D-glucosamine residues, is catalyzed by the enzyme glucosaminyl N-deacetylase/N-sulfotransferase (NDST). This event is a key step that regulates the overall sulfate content of the polysaccharide. Here, we report on the effects of NDST deficiency on Ca²⁺ kinetics in myotubes from NDST-1- and NDST-2-deficient mice, indicating a novel role for heparan sulfate in skeletal muscle physiology.

Immunostaining for specific heparan sulfate epitopes showed major changes in the heparan sulfate composition in skeletal muscle tissue derived from NDST-1^–/– mice and NDST^–/– cultured myotubes. Biochemical analysis indicates a relative decrease in both N-sulfation and 2-O-sulfation of skeletal muscle heparan sulfate. The core protein of heparan sulfate proteoglycan perlecan was not affected, as judged by immunohistochemistry. Also, acetylcholine receptor clustering and the occurrence of other ion channels involved in excitation-contraction coupling were not altered. In NDST-2^–/– mice and heterozygous mice no changes in heparan sulfate composition were observed. Using high-speed UV confocal laser scanning microscopy, aberrant Ca²⁺ kinetics were observed in NDST-1^–/– myotubes, but not in NDST-2^–/– or heterozygous myotubes. Electrically induced Ca²⁺ spikes had significantly lower amplitudes, and a reduced removal rate of cytosolic Ca²⁺, indicating the importance of heparan sulfate in muscle Ca²⁺ kinetics.

Key words: Acetylcholine receptor, Ca²⁺, Heparan sulfate, Sulfotransferase, Skeletal muscle

This article has been cited by other articles:

				I. Barbosa, C. Morin, S. Garcia, A. Duchesnay, M. Oudghir, G. Jenniskens, H.-Q. Miao, S. Guimond, G. Carpentier, J. Cebrian, et al. A synthetic glycosaminoglycan mimetic (RGTA) modifies natural glycosaminoglycan species during myogenesis J. Cell Sci., January 1, 2005; 118(1): 253 - 264. [Abstract] [Full Text] [PDF]

				J. Ledin, W. Staatz, J.-P. Li, M. Gotte, S. Selleck, L. Kjellen, and D. Spillmann Heparan Sulfate Structure in Mice with Genetically Modified Heparan Sulfate Production J. Biol. Chem., October 8, 2004; 279(41): 42732 - 42741. [Abstract] [Full Text] [PDF]

				N. C. Smits, A. A. Robbesom, E. M. M. Versteeg, E. M. A. van de Westerlo, P. N. R. Dekhuijzen, and T. H. van Kuppevelt Heterogeneity of Heparan Sulfates in Human Lung Am. J. Respir. Cell Mol. Biol., February 1, 2004; 30(2): 166 - 173. [Abstract] [Full Text] [PDF]