Application of polyacrylamide gel electrophoresis of fluorophore-labeled saccharides for analysis of hyaluronan and chondroitin sulfate in human and animal tissues and cell cultures.
Hyaluronan (HA) and chondroitin sulfate (CS) are glycosaminoglycans (GAGs) with great importance in biological events, since they participate in and regulate cell adhesion, migration and proliferation. Quantitation and analysis of the fine structure of GAGs are increasingly important for understanding many biological processes, among which are many critical aspects of pathology development and specific phenotype descriptions. Human umbilical cord and human synovial fluid are connective tissues containing high amounts of GAGs and change in the quantity and structure of these macromolecules is described in tissue development and is commonly associated with disease. Moreover, also in Xenopus laevis embryo development and chondrocyte cultures, the GAG content and structure play a critical role. A rapid analysis of hyaluronan and chondroitin sulfate Delta-disaccharides derived from the above human and animal samples, derivatized with 2-aminoacridone and analyzed by polyacrylamide gel electrophoresis, is described in this report. Qualitative and quantitative analysis were performed by comparing their migration and the pixel density with standard Delta-disaccharides, running in the same gel. Since this method allows the analysis of large numbers of samples simultaneously in one gel and has a relatively high sensitivity (less than 25 pmol), it is suggested as a cost-effective and useful tool for the fast screening of small amounts of hyaluronan and chondroitin sulfate disaccharides.
Karousou
Evgenia G
EG
Department of Experimental and Clinical Biomedical Sciences, University of Insubria, Varese, Italy.
Viola
Manuela
M
Genasetti
Anna
A
Vigetti
Davide
D
Luca
Giancarlo De
GD
Karamanos
Nikos K
NK
Passi
Alberto
A
D000609:Aminoacridines / Q000737:chemistry; D000818:Animals; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000032:analysis*; D003470:Culture Media / Q000737:chemistry; D004187:Disaccharides / Q000032:analysis* / Q000302:isolation & purification*; D004314:Down Syndrome / Q000196:embryology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005312:Fetal Blood / Q000737:chemistry; D006801:Humans; D006820:Hyaluronic Acid / Q000032:analysis*; D012680:Sensitivity and Specificity; D013582:Synovial Fluid / Q000737:chemistry; D014982:Xenopus laevis / Q000196:embryology
D000609:Aminoacridines; D003470:Culture Media; D004187:Disaccharides; C072515:2-aminoacridone; D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates