Functions of Chondroitin Sulfate/Dermatan Sulfate Chains in Brain Development

Abstract

Chondroitin sulfate (CS) and dermatan sulfate (DS) have been implicated in the processes of neural development in the brain. In this study, we characterized developmentally regulated brain CS/DS chains using a single chain antibody, GD3G7, produced by the phage display technique. Evaluation of the specificity of GD3G7 toward various glycosaminoglycan preparations showed that this antibody specifically reacted with squid CS-E (rich in the GlcUA␤1-3GalNAc(4,6-O-sulfate) disaccharide unit E), hagfish CS-H (rich in the IdoUA␣1-3GalNAc(4,6-Osulfate) unit iE), and shark skin DS (rich in both E and iE units). In situ hybridization for the expression of N-acetylgalactosamine-4-sulfate 6-O-sulfotransferase in the postnatal mouse brain, which is involved in the biosynthesis of CS/DS-E, showed a widespread expression of the transcript in the developing brain except at postnatal day 7, where strong expression was observed in the external granule cell layer in the cerebellum. The expression switched from the external to internal granule cell layer with development. Immunohistochemical localization of GD3G7 in the mouse brain showed that the epitope was relatively abundant in the cerebellum, hippocampus, and olfactory bulb. GD3G7 suppressed the growth of neurites in embryonic hippocampal neurons mediated by CS-E, suggesting that the epitope is embedded in the neurite outgrowth-promoting motif of CS-E. In addition, a CS-E decasaccharide fraction was found to be the critical minimal structure needed for recognition by GD3G7. Four discrete decasaccharide epitopic sequences were identified. The antibody GD3G7 has broad applications in investigations of CS/DS chains during the central nervous system's development and under various pathological conditions.

Structure of Pleiotrophin- and Hepatocyte Growth Factor-binding Sulfated Hexasaccharide Determined by Biochemical and Computational Approaches

“…The NOP activity of CS/DS chains is mediated by PTN and HGF (19,20), and depends on the presence of disulfated disaccharide units and IdoUA (13)(14)(15)(16)(17)(18)(19)(20). Previously, we isolated sev- (21).…”

“…The presence of oversulfated disaccharides (13)(14)(15)(16) and IdoUA are two crucial factors for these functions of CS/DS chains (19,20), indicating that IdoUA-containing oligosaccharide sequences with specific sulfation patterns may form functional domains for growth factor-binding and NOP activities. However, such functional sequences with a CS/DS hybrid structure are not fully understood.…”

“…Recent studies showed that CS/DS chains are involved in brain development (10,11), and CS/DS variants from marine organisms and embryonic pig brain display growth factor-binding and neurite outgrowth-promoting (NOP) activities in vitro (13)(14)(15)(16)(17)(18). The presence of oversulfated disaccharides (13)(14)(15)(16) and IdoUA are two crucial factors for these functions of CS/DS chains (19,20), indicating that IdoUA-containing oligosaccharide sequences with specific sulfation patterns may form functional domains for growth factor-binding and NOP activities.…”

A Peptide Found by Phage Display Discriminates a Specific Structure of a Trisaccharide in Heparin

“…Because almost no glycosaminoglycans are immunoreactive and there are few antibodies against them, this advantage is extremely useful for generating probes that bind to glycosaminoglycans. Thus, phage display technology is increasingly important for studying the functions of glycosaminoglycans (47)(48)(49)(50)(51)(52). The generation of type-specific anti-HS antibodies using phage display technology was initially applied by van Kuppevelt et al (53) for investigating HS heterogeneity in the kidney.…”

Development of a mouse monoclonal antibody against the chondroitin sulfate-protein linkage region derived from shark cartilage

“…Monoclonal antibodies (mAbs) that recognize specific features of the repeating disaccharide region of CS chains have been generated, and epitopes of some of them have been characterized [12][13][14][15][16]. Studies using anti-CS mAbs have revealed restricted spatiotemporal patterns of the expression of specific CS structures in various tissues during growth and development, and in pathological conditions [16][17][18][19]. However, so far, no antibodies have been reported which recognize the GAG-protein linkage region.…”