Chondroitin sulfate/dermatan sulfate hybrid chains in the development of cerebellum. Spatiotemporal regulation of the expression of critical disulfated disaccharides by specific sulfotransferases.
Chondroitin sulfate/dermatan sulfate (CS/DS) chains regulate the development of the central nervous system in vertebrates. Previously, we demonstrated that CS/DS hybrid chains from embryonic pig brain exhibit neuritogenic and growth factor binding activities, which depended on their IdoUA content defining the DS-like structure. To elucidate the distribution of such functional sugar chains during the development of the brain, in situ hybridization was performed to examine expression of three CS/DS GalNAc 4-O-sulfotransferases, D4ST-1, C4ST-1, and C4ST-2, and a single uronyl 2-O-sulfotransferase (UST) involved in the biosynthesis of DS in addition to CS intermediates. C4ST-1 and C4ST-2 were ubiquitously expressed in the postnatal mouse brain, whereas the expression of D4ST-1 and UST was restricted in the developing cerebellum and culminated at postnatal day 14 as shown by reverse transcriptase-PCR analysis. In situ analysis of the disaccharides of CS/DS in brain sections revealed that the concentration of CS/DS increases 2-fold during development (postnatal day 7 to 7 weeks). The proportions of DS-specific, principal disaccharides, IdoUA-Gal-NAc(4-O-sulfate) (iA) and IdoUA(2-O-sulfate)-GalNAc(4-O-sulfate) (iB), produced by the sequential actions of D4ST-1 and UST, were higher in the CS/DS chains from cerebellum than those from whole brain sections. A dramatic increase (10-fold) in the proportion of iB during development was noteworthy. In contrast, GlcUA/IdoUA(2-O-sulfate)-GalNAc(6-O-sulfate) (D/iD) and GlcUA/IdoUA-GalNAc(4, 6-O-disulfate) (E/iE) decreased to 50 and 30%, respectively, in the developing cerebellum. These results suggest that the IdoUA-containing iA and iB units along with D/iD and E/iE units in the CS/DS hybrid play important roles in the formation of the cerebellar neural network during postnatal brain development.
Mitsunaga
Chie
C
Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan.
Mikami
Tadahisa
T
Mizumoto
Shuji
S
Fukuda
Junko
J
Sugahara
Kazuyuki
K
D000818:Animals; D001921:Brain / Q000196:embryology* / Q000378:metabolism*; D002531:Cerebellum / Q000196:embryology*; D002809:Chondroitin Sulfates / Q000378:metabolism*; D003001:Cloning, Molecular; D003871:Dermatan Sulfate / Q000378:metabolism*; D004187:Disaccharides / Q000737:chemistry; D018507:Gene Expression Regulation, Developmental; D017403:In Situ Hybridization; D051379:Mice; D009844:Oligosaccharides / Q000737:chemistry; D015238:Sulfotransferases / Q000737:chemistry / Q000378:metabolism; D013552:Swine; D013997:Time Factors; D000097811:Carbohydrate Sulfotransferases
D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D004187:Disaccharides; D009844:Oligosaccharides; D015238:Sulfotransferases; C569944:chondroitin 4-O-sulfotransferase 2, mouse; D000097811:Carbohydrate Sulfotransferases; C000606136:uronyl 2-O-sulfotransferase, mouse