title: Chondroitin sulfate and keratan sulfate are the major glycosaminoglycans present in the adult zebrafish Danio rerio (Chordata-Cyprinidae).
abstract: The zebrafish Danio rerio (Chordata-Cyprinidae) is a model organism frequently used to study the functions of proteoglycans and their glycosaminoglycan (GAG) chains. Although several studies clearly demonstrate the participation of these polymers in different biological and cellular events that take place during embryonic development, little is known about the GAGs in adult zebrafish. In the present study, the total GAGs were extracted from the whole fish by proteolytic digestion, purified by anion-exchange chromatography and characterized by electrophoresis after degradation with specific enzymes and/or by high-performance liquid chromatography (HPLC) analysis of the disaccharides. Two GAGs were identified: a low-molecular-weight chondroitin sulfate (CS) and keratan sulfate (KS), corresponding to approximately 80% and 20% of the total GAGs, respectively. In the fish eye, KS represents approximately 80% of total GAGs. Surprisingly, no heparinoid was detected, but may be present in the fish at concentrations lower than the limit of the method used. HPLC of the disaccharides formed after chondroitin AC or ABC lyase degradation revealed that the zebrafish CS is composed by DeltaUA-1-->3-GalNAc(4SO4) (59.4%), DeltaUA-1-->3-GalNAc(6SO4) (23.1%), and DeltaUA-1-->3-GalNAc (17.5%) disaccharide units. No disulfated disaccharides were detected. Immunolocalization on sections from zebrafish retina using monoclonal antibodies against CS4- or 6-sulfate showed that in the retina these GAGs are restricted to the outer and inner plexiform layers. This is the first report showing the presence of KS in zebrafish eye, and the structural characterization of CS and its localization in the zebrafish retina. Detailed information about the structure and tissue localization of GAGs is important to understand the functions of these polymers in this model organism.
authors.affiliation: Laboratório de Tecido Conjuntivo, Hospital Universitário Clementino Fraga Filho and Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Caixa Postal 68041, Rio de Janeiro, RJ, CEP 21941-590, Brazil.
authors.forename: Aline R C
authors.initials: AR
authors.lastname: Souza
authors.forename: Eliene O
authors.initials: EO
authors.lastname: Kozlowski
authors.forename: Vinicius R
authors.initials: VR
authors.lastname: Cerqueira
authors.forename: Morgana T L
authors.initials: MT
authors.lastname: Castelo-Branco
authors.forename: Manoel L
authors.initials: ML
authors.lastname: Costa
authors.forename: Mauro S G
authors.initials: MS
authors.lastname: Pavão
mesh_terms: D000818:Animals; D002809:Chondroitin Sulfates / Q000737:chemistry* / Q000502:physiology; D003315:Cornea / Q000378:metabolism; D004187:Disaccharides / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006025:Glycosaminoglycans / Q000378:metabolism*; D006603:Hexuronic Acids / Q000737:chemistry; D007632:Keratan Sulfate / Q000737:chemistry* / Q000502:physiology; D008954:Models, Biological; D008970:Molecular Weight; D009608:Nitrous Acid / Q000737:chemistry; D012160:Retina / Q000378:metabolism; D014574:Uronic Acids / Q000737:chemistry; D015027:Zebrafish
chemical_list: D004187:Disaccharides; D006025:Glycosaminoglycans; D006603:Hexuronic Acids; D014574:Uronic Acids; D002809:Chondroitin Sulfates; D007632:Keratan Sulfate; D009608:Nitrous Acid