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1 Dept. of Nephrology, Clinical Sciences, Lund University, Lund, Sweden
* To whom correspondence should be addressed. E-mail: Bengt.Rippe{at}med.lu.se.
It is established that the glomerular filter sieves macromolecules based on their size, shape and charge. Anionic proteins are thus retarded compared to their neutral or cationic counterparts. However, recent studies have indicated that charge effects are small, or even 'anomalous' for polysaccharides. We therefore investigated the impact of charge on the glomerular permeability to polysaccharides by comparing sieving coefficients (
) (primary urine to plasma concentration ratio) for negatively charged, carboxymethylated (CM), FITC-Ficoll and FITC-dextran to their neutral counterparts. for these probes were determined in anaesthetized Wistar rats, 269±2.7 g (±SE; n=36), whose ureters were cannulated for urine sampling. The glomerular filtration rate was assessed using FITC-inulin. Polysaccharides were constantly infused, and after equilibration, urine was collected and a midpoint plasma sample taken. Size and concentration determinations of the FITC-labeled polysaccharides were achieved by size exclusion HPLC (HPSEC). For CM-Ficoll, was significantly increased (32 times at 55 A) compared to that of uncharged Ficoll. A small increase in for CM-dextran compared to neutral dextran was also observed (1.8 times at 55 A). In conclusion, negatively charged Ficoll relative to neutral Ficoll, was found to be markedly hyperpermeable across the glomerular filter. Furthermore, negatively charged Ficoll was observed to be larger on HPSEC compared to its neutral counterpart of same molecular weight. It is proposed that the introduction of negative charges in the dendrimeric, cross-linked, Ficoll molecule may alter its configuration, so as to make it more extended, and conceivably, more flexible, thereby increasing its glomerular permeability.
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