title: Chondroitin 4-O-sulfotransferase-1 is required for somitic muscle development and motor axon guidance in zebrafish.
abstract: CS (chondroitin sulfate) has been implicated in a variety of biological processes during development. Its biological functions are closely associated with characteristic sulfated structures. Here, we report the characterization of a zebrafish counterpart of C4ST-1 (chondroitin 4-O-sulfotransferase-1) and its functional importance in embryogenesis. Recombinant C4ST-1 showed a substrate preference for chondroitin and catalysed the 4-O-sulfation of GalNAc residues, a highly frequent modification of CS in the embryos of zebrafish as well as other vertebrates. Whole-mount in situ hybridization revealed that C4ST-1 showed a distinct spatiotemporal expression pattern in the developing zebrafish embryo. During the segmentation stages, strong expression was observed along the body axis including the notochord and somites. Functional knockdown of C4ST-1 with specific antisense morpholino-oligonucleotides led to a marked decrease in the 4-O-sulfation and amount of CS in the embryos. Consistent with the preferential expression in the rostrocaudal axis, C4ST-1 morphants displayed morphological defects exemplified by a ventrally bent trunk and a curled and/or kinky tail, largely due to misregulated myotomal myod expression, implying perturbation of axial muscle differentiation in somites. Furthermore, the aberrant projection of spinal motor axons, which extended ventrally at the interface between the notochord and individual somites, was also observed in C4ST-1 morphants. These results suggest that 4-O-sulfated CS formed by C4ST-1 is essential for somitic muscle differentiation and motor axon guidance in zebrafish development.
authors.affiliation: Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan.
authors.forename: Shuji
authors.initials: S
authors.lastname: Mizumoto
authors.forename: Tadahisa
authors.initials: T
authors.lastname: Mikami
authors.forename: Daiki
authors.initials: D
authors.lastname: Yasunaga
authors.forename: Naoki
authors.initials: N
authors.lastname: Kobayashi
authors.forename: Hajime
authors.initials: H
authors.lastname: Yamauchi
authors.forename: Ayumi
authors.initials: A
authors.lastname: Miyake
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Itoh
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kitagawa
authors.forename: Kazuyuki
authors.initials: K
authors.lastname: Sugahara
mesh_terms: D000818:Animals; D002809:Chondroitin Sulfates / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D024510:Muscle Development / Q000235:genetics*; D055495:Neurogenesis / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015027:Zebrafish / Q000196:embryology* / Q000378:metabolism*; D029961:Zebrafish Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
chemical_list: D007527:Isoenzymes; D029961:Zebrafish Proteins; D002809:Chondroitin Sulfates; D015238:Sulfotransferases; C019893:chondroitin 4-sulfotransferase