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J. Biol. Chem., Vol. 282, Issue 2, 843-852, January 12, 2007

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Heparan Sulfate Analysis from Diabetic Rat Glomeruli^*

From the Department of Biomedical Engineering, Cleveland Clinic, Cleveland, Ohio 44195

One of the major complicating factors in insulin-dependent diabetes mellitus is nephropathy. Several investigators have linked heparan sulfate (HS) alterations in the glomerular basement membrane (GBM) with albuminuria as a marker of abnormal blood filtration and the subsequent progression to renal failure. In this study, we examined the fine structure of HS in the glomerulus and the GBM isolated from the kidneys of rats injected with streptozotocin. Using fluorophore-assisted carbohydrate electrophoresis, we obtained disaccharide composition analyses for HS. In a time course study, we observed that normal rat HS isolated from the GBM becomes more N-sulfated as the glomeruli mature over a period of 8 weeks. Diabetic rats injected with streptozotocin at the beginning of this period showed a reversal of this trend. Using a graded sieve technique, we found that two different sizes of glomeruli could be isolated from the rat kidneys and that there was a significant difference in the HS disaccharide content between these two pools of glomeruli. Only the larger sized glomeruli had less N-sulfation of HS as a result of insulin-dependent diabetes mellitus. This change in the fine structure of HS was localized to the GBM and was not associated with cell surface HS. We also generated oligosaccharides of HS that portray fine structural alterations in the diabetic rats indicative of a loss of the sulfation of N-acetylglucosamine.

Received for publication, September 12, 2006

^* This work was supported by National Institutes of Health Grants K01 DK02847 and R01 DK62934 and Diabetes Association of Greater Cleveland Grant 453 (to A. W.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains a supplemental figure.

¹ To whom correspondence should be addressed: Lerner Research Institute, ND20, Dept. of Biomedical Engineering, Cleveland Clinic, Cleveland, OH 44195. Tel.: 216-445-3237; Fax: 216-444-9198; E-mail: wanga{at}ccf.org.

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