Document Detail


Improved and simple micro assay for sulfated glycosaminoglycans quantification in biological extracts and its use in skin and muscle tissue studies.
MedLine Citation:
PMID:  12773478     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
This article describes a simple and selective procedure used for direct measurement of sulfated glycosaminoglycans (GAGs) in biological samples and its application to the determination of GAGs during tissue regeneration and myogenic differentiation. We describe a modified procedure of previous GAG assays that has improved specificity, reproducibility, and sensitivity. The assay is based on the ability of sulfated GAGs to bind the cationic dye 1,9-dimethylmethylene blue. We describe conditions that allow isolation of the GAG-dye complex. This complex was dissociated; the optical density measurement of the dissociated dye permitted quantification of GAGs in biological samples. Applied to the study of myogenic cell differentiation in vitro, muscle repair, and skin ulceration, this method revealed significant modifications in the patterns of expression of different sulfated GAGs in these tissues. In particular, application of the method after nitrous acid treatment revealed that heparan sulfate and chondroitin sulfate ratio changed during muscle regeneration process.
Authors:
Isabelle Barbosa; Stéphanie Garcia; Véronique Barbier-Chassefière; Jean-Pierre Caruelle; Isabelle Martelly; Dulce Papy-García
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2003-05-28
Journal Detail:
Title:  Glycobiology     Volume:  13     ISSN:  0959-6658     ISO Abbreviation:  Glycobiology     Publication Date:  2003 Sep 
Date Detail:
Created Date:  2003-08-22     Completed Date:  2004-05-14     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9104124     Medline TA:  Glycobiology     Country:  England    
Other Details:
Languages:  eng     Pagination:  647-53     Citation Subset:  IM    
Affiliation:
Laboratoire de Recherche sur la Croissance, la Réparation et la Régénération Tissulaires (CRRET), CNRS FRE-2412, Université Paris 12-Val de Marne, 61 Avenue du Général de Gaulle, 94010 Créteil cedex, France.
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MeSH Terms
Descriptor/Qualifier:
Animals
Calibration
Cell Differentiation
Cell Line
Glycosaminoglycans / analysis*
Heparitin Sulfate
Male
Methylene Blue / analogs & derivatives*
Mice
Muscles / cytology,  metabolism*
Rats
Rats, Wistar
Regeneration
Reproducibility of Results
Sensitivity and Specificity
Skin / metabolism*,  pathology
Chemical
Reg. No./Substance:
0/Glycosaminoglycans; 0/dimethylmethylene blue; 61-73-4/Methylene Blue; 64082-61-7/A73025; 9050-30-0/Heparitin Sulfate

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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