Chondroitin / Dermatan Sulfate Modification Enzymes in Zebrafish Development

Abstract

Chondroitin/dermatan sulfate (CS/DS) proteoglycans consist of unbranched sulfated polysaccharide chains of repeating GalNAc-GlcA/IdoA disaccharide units, attached to serine residues on specific proteins. The CS/DS proteoglycans are abundant in the extracellular matrix where they have essential functions in tissue development and homeostasis. In this report a phylogenetic analysis of vertebrate genes coding for the enzymes that modify CS/DS is presented. We identify single orthologous genes in the zebrafish genome for the sulfotransferases chst7, chst11, chst13, chst14, chst15 and ust and the epimerase dse. In contrast, two copies were found for mammalian sulfotransferases CHST3 and CHST12 and the epimerase DSEL, named chst3a and chst3b, chst12a and chst12b, dsela and dselb, respectively. Expression of CS/DS modification enzymes is spatially and temporally regulated with a large variation between different genes. We found that CS/DS 4-O-sulfotransferases and 6-O-sulfotransferases as well as CS/DS epimerases show a strong and partly overlapping expression, whereas the expression is restricted for enzymes with ability to synthesize di-sulfated disaccharides. A structural analysis further showed that CS/DS sulfation increases during embryonic development mainly due to synthesis of 4-O-sulfated GalNAc while the proportion of 6-O-sulfated GalNAc increases in later developmental stages. Di-sulfated GalNAc synthesized by Chst15 and 2-O-sulfated GlcA/IdoA synthesized by Ust are rare, in accordance with the restricted expression of these enzymes. We also compared CS/DS composition with that of heparan sulfate (HS). Notably, CS/DS biosynthesis in early zebrafish development is more dynamic than HS biosynthesis. Furthermore, HS contains disaccharides with more than one sulfate group, which are virtually absent in CS/DS.

Knockdown of chondroitin‐4‐sulfotransferase‐1, but not of dermatan‐4‐sulfotransferase‐1, accelerates regeneration of zebrafish after spinal cord injury

“…Expression of Chst-11 is shifted from the notochord to the somites, hindbrain, midbrain, and forebrain at 24 h after fertilization. By 36 h after fertilization, the hindbrain, forebrain, and some cells of the midbrain and pectoral fin bud express Chst-11 (44). In the present study, Chst-11 MO knockdown at the 2-to 4-cell stage not only prompted spinal cord repair at the lesion site, but also accelerated functional recovery after SCI.…”

“…Analysis of CS and DS patterns during early zebrafish development showed that 4-O-sulfate groups are the most common modification, in accordance with the strong expression of CS and DS 4-O-sulfotransferases (44). Studies on zebrafish embryos had indicated that Chst-11 is strongly expressed along the body axis including the notochord and somites, whereas Chst-14 predominates in the head region and midbrain-hindbrain boundary (25,44).…”

“…We cannot explain this phenomenon because expression of CS/DS-modifying enzymes during zebrafish embryonic development and in the adult has not been studied systematically. However, we would like to refer to a study on the phylogeny and expression of all known CS/DSmodifying enzymes as well as the temporal variation of CS/DS structures during development (44). This study explains a few points that may enable better understanding of the phenomenon that, unlike their function in the mouse, CS/DS biosynthetic enzymes produce tissue-specific structures during zebrafish development and CS/DS disaccharides with multiple sulfate groups that are rare in zebrafish.…”

Reconstruction of the carbohydrate 6-O sulfotransferase gene family evolution in vertebrates reveals novel member, CHST16, lost in amniotes

“…in the maintenance of cartilaginous tissue (Hayashi et al 2011). The expression profiles of 812 the zebrafish chst3a, chst3b and chst7 genes during development mirror the known functions 813 of CHST3 and CHST7 in mammals, with expression mainly in skeletal tissues and the central 814 nervous system (Habicher et al 2015), albeit with only a very weak signal for chst3b. Further 815 study in teleost species should be done in order to compare the expression profiles of the 816 teleost-specific CHST3 duplicates.…”

Comparative Huntington and Parkinson Disease mRNA Analysis Reveals Common Inflammatory Processes

“…The most consistently ranked gene is RP1-93H18.7 (ENSG00000272403.1), a lncRNA, which was removed from Ensembl starting at version GRCh38.p2, but shows consistent transcription in these data. This gene is directly downstream of the gene DSE (dermatan sulfate epimerase) which is also DE in both HD and PD, is involved in embryonic development 20,21 , and has been related to the immune response in cancer patients 22 . Deficiencies in the second ranked gene, SPR (sepiapterin reductase), have been linked to DOPA-responsive dystonia 23 and previously implicated in PD 24 .…”