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Chondroitin / dermatan sulfate modification enzymes in zebrafish development.

Habicher J, Haitina T, Eriksson I, Holmborn K, Dierker T, Ahlberg PE, Ledin J - PLoS ONE (2015)

Bottom Line: 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.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.We also compared CS/DS composition with that of heparan sulfate (HS).

View Article: PubMed Central - PubMed

Affiliation: Department of Organismal Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.

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.

No MeSH data available.


Whole mount in situ hybridization of the DS epimerases dse (A), dsela (B) and dselb (C).All images show lateral views, if not otherwise stated in the figure. hg: hatching gland, MBH: midbrain-hindbrain boundary, p: polster, pc: pharyngeal cartilage, pf: pectoral fin, s: somites
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pone.0121957.g006: Whole mount in situ hybridization of the DS epimerases dse (A), dsela (B) and dselb (C).All images show lateral views, if not otherwise stated in the figure. hg: hatching gland, MBH: midbrain-hindbrain boundary, p: polster, pc: pharyngeal cartilage, pf: pectoral fin, s: somites

Mentions: Both probes for dse and dselb stain at the 2-cell stage (¾ hpf), indicating maternal deposition of mRNA (Fig. 6A,C) while no staining was seen at this stage for dsela (Fig. 6B). A general weak staining of dse is detected at the 5- and 15-somite stage (11–16 hpf) while somites and the eyes stain strongly (Fig. 6A). At 24 and 36 hpf, only the posterior end of the somites remain stained and staining appears strong in the most ventral regions of the head (Fig. 6A). At 72 hpf staining is still strong and general in the most ventral regions of the head and pectoral fins (Fig. 6A). dsela stains very strong in the polster at the 5-somite stage and shows clear staining in cells of the hatching gland at the 15-somite stage (Fig. 6B). The expression in the hatching gland remains persistent at 24, 36 and 48 hpf, while no staining is detected at 72 hpf (Fig. 6B). dselb shows a general expression at the somite stages, and at 24 hpf, somites as well as the head region and the hatching gland are stained (Fig. 6C). At 36 and 48 hpf, in addition to the hatching gland, also the midbrain-hindbrain boundary and the pectoral fins are strongly labeled (Fig. 6C). By 72 hpf, staining is weak and restricted to the pectoral fins and the cartilage elements (Fig. 6C).


Chondroitin / dermatan sulfate modification enzymes in zebrafish development.

Habicher J, Haitina T, Eriksson I, Holmborn K, Dierker T, Ahlberg PE, Ledin J - PLoS ONE (2015)

Whole mount in situ hybridization of the DS epimerases dse (A), dsela (B) and dselb (C).All images show lateral views, if not otherwise stated in the figure. hg: hatching gland, MBH: midbrain-hindbrain boundary, p: polster, pc: pharyngeal cartilage, pf: pectoral fin, s: somites
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4368567&req=5

pone.0121957.g006: Whole mount in situ hybridization of the DS epimerases dse (A), dsela (B) and dselb (C).All images show lateral views, if not otherwise stated in the figure. hg: hatching gland, MBH: midbrain-hindbrain boundary, p: polster, pc: pharyngeal cartilage, pf: pectoral fin, s: somites
Mentions: Both probes for dse and dselb stain at the 2-cell stage (¾ hpf), indicating maternal deposition of mRNA (Fig. 6A,C) while no staining was seen at this stage for dsela (Fig. 6B). A general weak staining of dse is detected at the 5- and 15-somite stage (11–16 hpf) while somites and the eyes stain strongly (Fig. 6A). At 24 and 36 hpf, only the posterior end of the somites remain stained and staining appears strong in the most ventral regions of the head (Fig. 6A). At 72 hpf staining is still strong and general in the most ventral regions of the head and pectoral fins (Fig. 6A). dsela stains very strong in the polster at the 5-somite stage and shows clear staining in cells of the hatching gland at the 15-somite stage (Fig. 6B). The expression in the hatching gland remains persistent at 24, 36 and 48 hpf, while no staining is detected at 72 hpf (Fig. 6B). dselb shows a general expression at the somite stages, and at 24 hpf, somites as well as the head region and the hatching gland are stained (Fig. 6C). At 36 and 48 hpf, in addition to the hatching gland, also the midbrain-hindbrain boundary and the pectoral fins are strongly labeled (Fig. 6C). By 72 hpf, staining is weak and restricted to the pectoral fins and the cartilage elements (Fig. 6C).

Bottom Line: 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.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.We also compared CS/DS composition with that of heparan sulfate (HS).

View Article: PubMed Central - PubMed

Affiliation: Department of Organismal Biology, Science for Life Laboratory, Uppsala University, Uppsala, Sweden.

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.

No MeSH data available.