PMID: 8947501Nov 15, 1996Paper

A steady-state labelling approach to the measurement of proteoglycan turnover in vivo and its application to glomerular proteoglycans

The Biochemical Journal
E L AkuffoR M Mason

Abstract

Rats were implanted with mini osmotic pumps delivering sodium [35S]sulphate and their newly synthesized proteoglycans were labelled over a 146 h period (steady-state labelling). Proteoglycan turnover was measured in vivo using a chase protocol. Glomerular proteoglycans were recovered quantitatively and the perlecan present was isolated by immunoprecipitation. The procedure allows newly synthesized proteoglycans to be quantified in mass units (pmol of glycossminoglycan sulphate) after labelling and during the chase. Ultrastructural-immunogold experiments identified the location of perlecan as the glomerular basement membrane and mesangial matrix. Perlecan in the basement membrane was quantified using the ultrastructural-immunogold technique. Perlecan comprises about 10% of the total glomerular proteoglycans, which are otherwise associated with glomerular cells and the mesangium. Both the total glomerular heparan sulphate proteoglycans and perlecan turn over rapidly (t1/2 approximately 3-4 h and < 3 h respectively). In contrast, turnover of proteoglycans in other tissues was slow, except in the liver where the heparan sulphate and chondroitin sulphate t1/2 values were 16 h and 9 h respectively. Microalbuminuria was induced with a...Continue Reading

Citations

May 26, 2001·Proceedings. Biological Sciences·D Fernández-MosqueraA Grandal-d'Anglade
Nov 3, 2016·American Journal of Physiology. Renal Physiology·Caroline B Marshall
Oct 22, 2002·American Journal of Physiology. Renal Physiology·Jenny SörenssonBörje Haraldsson

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