Effect of polycations on permeability of glomerular epithelial cell monolayers to albumin.
Polycations can interact with the surface negative charges of the glomerular epithelial cells in the kidney and give rise to metabolic alterations. This study examined whether charge neutralization can affect intercellular junctions and increase macromolecular permeability across epithelial monolayers. We examined this question by studying the effect of polycations on the leakage of albumin across monolayers of glomerular epithelial cells. Cells were grown to confluency on filter-lined cups. They were treated apically with cationic bovine gamma globulin or protamine (100 micrograms/ml) for 2 hours at 37 degrees C. After washing the cells, the monolayers were tested for leakage of albumin by the addition of radioactive bovine serum albumin on the apical side and determining the time course of its appearance on the basal side. Polycation treatment caused significant leakage of albumin in the absence of any toxic effect on viability or lactate dehydrogenase release. The leakage was shown to be through the tight junctions of the monolayer. Permeability alterations were compared at 4 degrees C and 37 degrees C to determine whether impairment was due to the membrane ruffling effect of charge neutralization or due to intracellular metabolic changes. Despite equal bindings of polycations at 4 degrees C and 37 degrees C, significant leak occurred only at 37 degrees C, suggesting the role of active processes in the maintenance of permselectivity. This was consistent with the rapid interiorization of the polycation at 37 degrees C after membrane binding. Further substantiation of this point was obtained by studying the protective effect of removing bound polycation with heparin. Removal of polycation after initial binding failed to protect the monolayer from albumin leakage. The conclusion was that neutralization of glomerular epithelial cell surface charges results in subtle impairment of tight junction control of permeability to macromolecules across the monolayer. The impairment was due to active intracellular processes that ensue after polycation binding to cell surface charges and their subsequent internalization.
Hammes
M
M
Division of Renal Disease and Hypertension, Loyola-Hines Medical Center, IL.
Singh
A
A
D000818:Animals; D002412:Cations / Q000494:pharmacology*; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D004563:Electrochemistry; D004705:Endocytosis; D004847:Epithelial Cells; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D007365:Intercellular Junctions / Q000187:drug effects; D007678:Kidney Glomerulus / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D051381:Rats; D012710:Serum Albumin, Bovine / Q000493:pharmacokinetics*; D005719:gamma-Globulins / Q000493:pharmacokinetics
D002412:Cations; D005719:gamma-Globulins; D012710:Serum Albumin, Bovine