Structural and functional changes of sulfated glycosaminoglycans in Xenopus laevis during embryogenesis.
Xenopus laevis is an excellent animal for analyzing early vertebrate development. Various effects of glycosaminoglycans (GAGs) on growth factor-related cellular events during embryogenesis have been demonstrated in Xenopus. To elucidate the relationship between alterations in fine structure and changes in the specificity of growth factor binding during Xenopus development, heparan sulfate (HS) and chondroitin/dermatan sulfate (CS/DS) chains were isolated at four different embryonic stages and their structure and growth factor-binding capacities were compared. The total amounts of both HS and CS/DS chains decreased from the pre-midblastula transition to the gastrula stage, but increased exponentially during the following developmental stages. The length of HS chains was not significantly affected by development, whereas that of CS/DS chains increased with development. The disaccharide composition of GAGs in embryos also changed during development. The degree of sulfation of the HS chains gradually decreased with development. The predominant sulfation position in the CS/DS chains shifted from C4 to C6 of GalNAc during embryogenesis. Growth factor-binding experiments using a BIAcore system demonstrated that GAGs bound growth factors including fibroblast growth factors-1 and -2, midkine, and pleiotrophin, with comparable affinities. These affinities significantly varied during development, although the correlation between the structural alterations of GAGs and the change in the ability to bind growth factors remains to be clarified. The expression of saccharide sequences, which specifically interact with a growth factor, might be regulated during development.
Yamada
Shuhei
S
Laboratory of Proteoglycan Signaling and Therapeutics, Hokkaido University Graduate School of Life Science, Sapporo 001-0021, Japan. tjohej@sci.hokudai.ac.jp
Onishi
Masako
M
Fujinawa
Reiko
R
Tadokoro
Yuko
Y
Okabayashi
Koji
K
Asashima
Makoto
M
Sugahara
Kazuyuki
K
D000818:Animals; D002809:Chondroitin Sulfates; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003871:Dermatan Sulfate / Q000378:metabolism; D004186:Disaccharidases / Q000032:analysis* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D014982:Xenopus laevis / Q000196:embryology / Q000378:metabolism*
D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004186:Disaccharidases; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D036341:Intercellular Signaling Peptides and Proteins