Biosynthesis and metabolism of sulfated glycosaminoglycans during Drosophila melanogaster development

We developed a simple methodology for labeling sulfated glycosaminoglycans (GAGs) in adult Drosophila melanogaster and studied some aspects of the biosynthesis and metabolism of these polymers during development. Adult D. melanogaster flies were fed with Na235SO4 for 72 h. During this period, 35S-su...

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Published in:	Glycobiology (Oxford) Vol. 14; no. 6; pp. 529 - 536
Main Authors:	Pinto, Daniela O., Ferreira, Paola L., Andrade, Leonardo R., Petrs-Silva, Hilda, Linden, Rafael, Abdelhay, Eliana, Araújo, Helena M. M., Alonso, Carlos-Eloy V., Pavão, Mauro S.G.
Format:	Journal Article
Language:	English
Published:	England Oxford University Press 01-06-2004 Oxford Publishing Limited (England)
Subjects:	5 unsaturated hexuronic acid Animals biosynthesis chondroitin sulfate dermatan sulfate development Drosophila melanogaster Drosophila melanogaster - embryology Drosophila melanogaster - growth & development Drosophila melanogaster - metabolism EDTA Electrophoresis, Agar Gel ethylenediamine tetra-acetic acid GAG glycosaminoglycan Glycosaminoglycans - biosynthesis Glycosaminoglycans - metabolism heparan sulfate high-performance liquid chromatography HPLC Immunohistochemistry PBS phosphate buffered saline ΔUA
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Summary:	We developed a simple methodology for labeling sulfated glycosaminoglycans (GAGs) in adult Drosophila melanogaster and studied some aspects of the biosynthesis and metabolism of these polymers during development. Adult D. melanogaster flies were fed with Na235SO4 for 72 h. During this period, 35S-sulfate was incorporated into males and females and used to synthesize 35S-sulfate–heparan sulfate (HS) and 35S-sulfate–chondroitin sulfate (CS). The incorporation of 35S-sulfate into HS was higher when compared to CS. In a pulse-chase experiment, we observed that 35S-sulfate incorporated into adult female was recovered in embryos and used for the synthesis of new 35S-sulfate-GAGs after 2 h of embryonic development. The synthesis of CS was higher than that of HS, indicating a change in the metabolism of these glycans from adult to embryonic and larval stages. Analysis of the CS in embryonic and larval tissues revealed the occurrence of nonsulfated and 4-sulfated disaccharide units in embryos, L1 and L2. In L3, in addition to these disaccharides, we also detected significant amount of 6-sulfated units that are reported here for the first time. Immunohistochemical analysis indicated that HS and CS were present in nonequivalent structures in adult and larval stages of the fly. Overall, these results indicate that 35S-sulfate-precursors are transferred from adult to embryonic and larval tissues and used to assemble different morphological structures during development.
Bibliography:	local:cwh070 istex:D76C8578A6318E6EE7AB0DACB93246E7FB1141FD ark:/67375/HXZ-QXRRGRKP-2 1To whom correspondence should be addressed; e-mail: mpavao@hucff.ufrj.br ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0959-6658 1460-2423 1460-2423
DOI:	10.1093/glycob/cwh070