Glomerular charge selectivity for horseradish peroxidase and albumin at low and normal ionic strengths

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Abstract
The classical concept of a negative glomerular charge barrier has recently been questioned, mainly based on the somewhat high clearance for anionic horseradish peroxidase (HRP). The validity of using anionic HRP can be tested by changing the properties of the charge barrier. A rather unequivocal approach is to reduce the ionic composition of the perfusate and hence increase the Debye length. We determined the glomerular clearance for horseradish peroxidase and serum albumin, using isolated rat kidneys perfused at 8 °C to reduce the tubular modification of the primary urine. The perfusate contained trace amounts of the neutral 125I-nHRP and the anionic 131I-aHRP and were otherwise identical except for different ionic strengths, 152 mM and 34 mM, respectively. During control, the fractional clearance was 0.11 ± 0.015 for nHRP and 0.045 ± 0.010 for aHRP, with an average clearance ratio (n/a) of 2.8 ± 0.24. Low ionic strength reduced for aHRP to 0.027 ± 0.006, giving an increased clearance ratio for HRP of 4.2 ± 0.44. The existence of a negative charge barrier is supported by the experiments. The result obtained during normal perfusion is compatible with a charge density (ω) of 34 mEq L−1, using a model of homogeneously charged membrane. Low ionic strength perfusion reversibly reduced the concentration of fixed charges to 12 mEq L−1, suggesting an almost threefold increase of the glomerular membrane volume. Thus, the glomerular charge barrier should be regarded to have a dynamic gel structure rather than being a rigid membrane.

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