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Chondroitin sulfate and keratan sulfate are the major glycosaminoglycans present in the adult zebrafish Danio rerio (Chordata-Cyprinidae)

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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 ∼80% and 20% of the total GAGs, respectively. In the fish eye, KS represents ∼ 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 ΔUA-1→3-GalNAc(4SO4) (59.4%), ΔUA-1→3-GalNAc(6SO4) (23.1%), and ΔUA-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.

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Abbreviations

α-ΔUA(2SO4):

α-Δ4,5 unsaturated hexuronic acid 2-sulfate

α-ΔUA:

α-Δ4,5 unsaturated hexuronic acid

GalNAc(4SO4):

GalNAc(6SO4) and GalNAc(2,4-diSO4), derivatives of N-acetyl-d-galactosamine, bearing a sulfate ester at position 4, at position 6, and at both positions 2 and 4, respectively

HPLC:

high-performance liquid chromatography

FPLC:

fast protein liquid chromatography

References

  1. Kjellen, L., Lindahl, U.: Proteoglycans: structures and interactions. Annu. Rev. Biochem. 60, 443–475 (1991)

    Article  PubMed  CAS  Google Scholar 

  2. Iozzo, R.V.: Proteoglycans: Structure, Biology and Molecular Interactions. Marcel Dekker, Inc., New York (1998)

    Google Scholar 

  3. Esko, J.D., Selleck, S.B.: Order out of chaos: assembly of ligand binding sites in heparan sulfate. Annu. Rev. Biochem. 71, 435–471 (2002)

    Article  PubMed  CAS  Google Scholar 

  4. Suzuki, M.: Prosthetic group of cornea mucoid. J. Biochem. 30, 185–191 (1939)

    CAS  Google Scholar 

  5. Meyer, K., Linker, A., Davidson, E.A., Weissman, B.: The mucopolysaccharides of the cornea. J. Biol. Chem. 205, 611–616 (1953)

    PubMed  CAS  Google Scholar 

  6. Funderburgh, J.L.: Keratan sulfate: structure, biosynthesis, and function. Glycobiology 10, 951–958 (2000)

    Article  PubMed  CAS  Google Scholar 

  7. Busch-Nentwich, E., Sollner, C., Roehl, H., Nicolson, T.: The deafness gene dfna5 is crucial for ugdh expression and HA production in the developing ear in zebrafish. Development 131, 943–951 (2004)

    Article  PubMed  CAS  Google Scholar 

  8. Walsh, E.C., Stainier, D.Y.: UDP-glucose dehydrogenase required for cardiac valve formation in zebrafish. Science 293, 1670–1673 (2001)

    Article  PubMed  CAS  Google Scholar 

  9. Norton, W.H., Ledin, J., Grandel, H., Neumann, C.J.: HSPG synthesis by zebrafish Ext2 and Extl3 is required for Fgf10 signalling during limb development. Development 132, 4963–4973 (2005)

    Article  PubMed  CAS  Google Scholar 

  10. Lee, J.S., von der Hardt, S., Rusch, M.A., Stringer, S.E., Stickney, H.L., Talbot, W.S., Geisler, R., Nusslein-Volhard, C., Selleck, S.B., Chien, C.B., Roehl, H.: Axon sorting in the optic tract requires HSPG synthesis by ext2 (dackel) and extl3 (boxer). Neuron 44, 947–960 (2004)

    Article  PubMed  CAS  Google Scholar 

  11. Chen, E., Stringer, S.E., Rusch, M.A., Selleck, S.B., Ekker, S.C.: A unique role for 6-O sulfation modification in zebrafish vascular development. Dev. Biol. 284, 364–376 (2005)

    Article  PubMed  CAS  Google Scholar 

  12. Bink, R.J., Habuchi, H., Lele, Z-., Dolk, E., Joore, J., Rauch, G.J., Geisler, R., Wilson, S.W., den Hertog, J., Kimata, K., Zivkovic, D.: Heparan sulfate 6-O-sulfotransferase is essential for muscle development in zebrafish. J. Biol. Chem. 278, 31118–31127 (2003)

    Article  PubMed  CAS  Google Scholar 

  13. De Cat, B., Muyldermans, S.Y., Coomans, C., Degeest, G., Vanderschueren, B., Creemers, J., Biemar, F., Peers, B., David, G.: Processing by proprotein convertases is required for glypican-3 modulation of cell survival, Wnt signaling, and gastrulation movements. J. Cell. Biol. 163, 625–635 (2003)

    Article  PubMed  CAS  Google Scholar 

  14. Topczewski, J., Sepich, D.S., Myers, D.C., Walker, C., Amores, A., Lele, Z., Hammerschmidt, M., Postlethwait, J., Solnica-Krezel, L.: The zebrafish glypican knypek controls cell polarity during gastrulation movements of convergent extension. Dev. Cell 1, 251–264 (2001)

    Article  PubMed  CAS  Google Scholar 

  15. Chen, E., Hermanson, S., Ekker, S.C.: Syndecan-2 is essential for angiogenic sprouting during zebrafish development. Blood 103, 1710–1719 (2004)

    Article  PubMed  CAS  Google Scholar 

  16. Bernhardt, R.R., Schachner, M.: Chondroitin sulfates affect the formation of the segmental motor nerves in zebrafish embryos. Dev. Biol. 221, 206–219 (2000)

    Article  PubMed  CAS  Google Scholar 

  17. Bernhardt, R.R., Goerlinger, S., Roos, M., Schachner, M.: Anterior–posterior subdivision of the somite in embryonic zebrafish: implications for motor axon guidance. Dev. Dyn. 213, 334–347 (1998)

    Article  PubMed  CAS  Google Scholar 

  18. Becker, C.G., Becker, T.: Repellent guidance of regenerating optic axons by chondroitin sulfate glycosaminoglycans in zebrafish. J. Neurosci. 22, 842–853 (2002)

    PubMed  CAS  Google Scholar 

  19. Nandini, C.D., Mikami, T., Ohta, M., Itoh, N., Akiyama-Nambu, F., Sugahara, K.: Structural and functional characterization of oversulfated chondroitin sulfate/dermatan sulfate hybrid chains from the notochord of hagfish. Neuritogenic and binding activities for growth factors and neurotrophic factors. J. Biol. Chem. 279, 50799–50809 (2004)

    Article  PubMed  CAS  Google Scholar 

  20. Hayashi, N., Mizusaki, M.J., Kamei, K., Harada, S., Miyata, S.: Chondroitin sulfate proteoglycan phosphacan associates with parallel fibers and modulates axonal extension and fasciculation of cerebellar granule cells. Mol. Cell Neurosci. 30, 364–377 (2005)

    Article  PubMed  CAS  Google Scholar 

  21. Hikino, M., Mikami, T., Faissner, A., Vilela-Silva, A.C., Pavao, M.S., Sugahara, K.: Oversulfated dermatan sulfate exhibits neurite outgrowth-promoting activity toward embryonic mouse hippocampal neurons: implications of dermatan sulfate in neuritogenesis in the brain. J. Biol. Chem. 278, 43744–43754 (2003)

    Article  PubMed  CAS  Google Scholar 

  22. Bao, X., Pavao, M.S., Santos, J.C., Sugahara, K.: A functional dermatan sulfate epitope containing iduronate(2-O-sulfate)alpha1-3GalNAc(6-O-sulfate) disaccharide in the mouse brain: demonstration using a novel monoclonal antibody raised against dermatan sulfate of ascidian Ascidia nigra. J. Biol. Chem. 280, 23184–23193 (2005)

    Article  PubMed  CAS  Google Scholar 

  23. Cardoso, L.E., Mourao, P.A.: Glycosaminoglycan fractions from human arteries presenting diverse susceptibilities to atherosclerosis have different binding affinities to plasma LDL. Arterioscler. Thromb. 14, 115–124 (1994)

    PubMed  CAS  Google Scholar 

  24. Gandra, M., Cavalcante, M., Pavao, M.: Anticoagulant sulfated glycosaminoglycans in the tissues of the primitive chordate Styela plicata (Tunicata). Glycobiology 10, 1333–1340 (2000)

    Article  PubMed  CAS  Google Scholar 

  25. Farndale, R.W., Buttle, D.J., Barrett, A.J.: Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim. Biophys. Acta. 883, 173–177 (1986)

    PubMed  CAS  Google Scholar 

  26. Dietrich, C.P., Dietrich, S.M.: Electrophoretic behaviour of acidic mucopolysaccharides in diamine buffers. Anal. Biochem. 70, 645–647 (1976)

    Article  PubMed  CAS  Google Scholar 

  27. Bitter, T., Muir, H.M.: A modified uronic acid carbazole reaction. Anal. Biochem. 4, 330–334 (1962)

    Article  PubMed  CAS  Google Scholar 

  28. Shively, J.E., Conrad, H.E.: Formation of anhydrosugars in the chemical depolymerization of heparin. Biochemistry 15, 3932–3942(1976)

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from CNPq, FAPERJ and the NIH Fogarty International Center (R03 TW05775). Mauro S.G. Pavão and Manoel L. Costa are research fellows from CNPq.

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Authors and Affiliations

  1. 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

    Aline R. C. Souza, Eliene O. Kozlowski, Vinicius R. Cerqueira & Mauro S. G. Pavão

  2. Departamento de Histologia e Embriologia, Instituto de Ciências Biomédicas, UFRJ, Rio de Janeiro, RJ, CEP 21941-290, Brazil

    Morgana T. L. Castelo-Branco & Manoel L. Costa

Authors
  1. Aline R. C. Souza
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  2. Eliene O. Kozlowski
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  3. Vinicius R. Cerqueira
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  4. Morgana T. L. Castelo-Branco
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  5. Manoel L. Costa
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  6. Mauro S. G. Pavão
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Correspondence to Mauro S. G. Pavão.

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The contributions of Aline R.C. Souza and Eliene O. Kozlowski should be considered equal.

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Souza, A.R.C., Kozlowski, E.O., Cerqueira, V.R. et al. Chondroitin sulfate and keratan sulfate are the major glycosaminoglycans present in the adult zebrafish Danio rerio (Chordata-Cyprinidae). Glycoconj J 24, 521–530 (2007). https://doi.org/10.1007/s10719-007-9046-z

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  • DOI: https://doi.org/10.1007/s10719-007-9046-z

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