title: A potential role for chondroitin sulfate/dermatan sulfate in arm regeneration in Amphiura filiformis.
abstract: Glycosaminoglycans (GAGs), such as chondroitin sulfate (CS) and dermatan sulfate (DS) from various vertebrate and invertebrate sources are known to be involved in diverse cellular mechanisms during repair and regenerative processes. Recently, we have identified CS/DS as the major GAG in the brittlestar Amphiura filiformis, with high proportions of di- and tri-O-sulfated disaccharide units. As this echinoderm is known for its exceptional regeneration capacity, we aimed to explore the role of these GAG chains during A. filiformis arm regeneration. Analysis of CS/DS chains during the regeneration process revealed an increase in the proportion of the tri-O-sulfated disaccharides. Conversely, treatment of A. filiformis with sodium chlorate, a potent inhibitor of sulfation reactions in GAG biosynthesis, resulted in a significant reduction in arm growth rates with total inhibition at concentrations higher than 5 mM. Differentiation was less impacted by sodium chlorate exposure or even slightly increased at 1-2 mM. Based on the structural changes observed during arm regeneration we identified chondroitin synthase, chondroitin-4-O-sulfotransferase 2 and dermatan-4-O-sulfotransferase as candidate genes and sought to correlate their expression with the expression of the A. filiformis orthologue of bone morphogenetic factors, AfBMP2/4. Quantitative amplification by real-time PCR indicated increased expression of chondroitin synthase and chondroitin-4-O-sulfotransferase 2, with a corresponding increase in AfBMP2/4 during regeneration relative to nonregenerating controls. Our findings suggest that proper sulfation of GAGs is important for A. filiformis arm regeneration and that these molecules may participate in mechanisms controlling cell proliferation.
authors.affiliation: Department of Medical Biochemistry and Microbiology, The Biomedical Center, Uppsala University, Box 582, SE-751 23 Uppsala, Sweden.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Ramachandra
authors.affiliation: Department of Medical Biochemistry and Microbiology, The Biomedical Center, Uppsala University, Box 582, SE-751 23 Uppsala, Sweden.
authors.forename: Ramesh B
authors.initials: RB
authors.lastname: Namburi
authors.affiliation: Department of Biological and Environmental Sciences, University of Gothenburg, Sven Lovén Centre for Marine Sciences, Kristineberg, SE 451 78 Fiskebäckskil, Sweden.|Department of Integrative Biology, University of California, 3040 Valley Life Sciences Building, CA 94720-3140, Berkeley, USA.
authors.forename: Sam T
authors.initials: ST
authors.lastname: Dupont
authors.affiliation: Department of Biological and Environmental Sciences, University of Gothenburg, Sven Lovén Centre for Marine Sciences, Kristineberg, SE 451 78 Fiskebäckskil, Sweden.
authors.forename: Olga
authors.initials: O
authors.lastname: Ortega-Martinez
authors.affiliation: Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Centre, PO Box 9101, 6500 HB Njimegen, The Netherlands.
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
authors.affiliation: Department of Medical Biochemistry and Microbiology, The Biomedical Center, Uppsala University, Box 582, SE-751 23 Uppsala, Sweden.
authors.forename: Ulf
authors.initials: U
authors.lastname: Lindahl
authors.affiliation: Department of Medical Biochemistry and Microbiology, The Biomedical Center, Uppsala University, Box 582, SE-751 23 Uppsala, Sweden.
authors.forename: Dorothe
authors.initials: D
authors.lastname: Spillmann
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000235:genetics; D002704:Chlorates / Q000494:pharmacology; D002809:Chondroitin Sulfates / Q000096:biosynthesis* / Q000235:genetics; D003871:Dermatan Sulfate / Q000096:biosynthesis* / Q000235:genetics; D004187:Disaccharides / Q000235:genetics / Q000378:metabolism; D004447:Echinodermata / Q000235:genetics / Q000254:growth & development; D006025:Glycosaminoglycans / Q000096:biosynthesis* / Q000235:genetics; D012038:Regeneration / Q000235:genetics*; D015238:Sulfotransferases / Q000235:genetics
chemical_list: D002704:Chlorates; D004187:Disaccharides; D006025:Glycosaminoglycans; D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates; C466772:chondroitin-4-O-sulfotransferase-3; D015238:Sulfotransferases; C438013:dermatan-4-sulfotransferase-1; C019893:chondroitin 4-sulfotransferase; C032706:sodium chlorate
keywords: Amphiura filiformis; brittlestar; chondroitin sulfate; dermatan sulfate; regeneration