Cationized ferritin binding to anionic surfaces in normal and aminonucleoside nephrotic kidneys

The distribution and effects of polycationized ferritin (PCF) bound ot anionic sites on living plasmalemmas and basal laminae of normal and puromycin aminonucleoside nephrotic (PAN) kidneys were studied using an in vitro model system. Immersion of normal glomeruli in physiological saline solutions containing low concentrations of PCF (0.01 to 0.1 mg/ml) for 10 seconds results in preferential binding of PCF to microvillous projections on the glomerular epithelium (i.e., podocytes). Exposure to higher concentrations of PCF (1.0 mg/ml) for 10 seconds results in several layers of PCF distributed evenly over the urinary aspect of epithelial podocytes. In these short treatment times, the thin slit diaphragms which span the filtration slits appear to be impermeable to PCF. Within several minutes after PCF treatment, ferritin is found within caveolae on the surface of epithelial podocytes and within numerous pinosomes and larger endocytic vesicles within these cells. Longer treatment with PCF results in the narrowing of filtration slit spaces and the formation of junctions between adjacent podocyte foot processes. Occurringg coincident with these structural changes is a gradual accumulation of PCF in regular patches along the lamina rara externa (LRE) of the glomerular basement membrane. Loss of foot processes and accumulation of PCF in the LRE are prevented by treatment with either cytochalasin B (25 micro g/ml), D (2 micro g/ml) or incubation at low temperatures (0-4 degrees C). When PCF-coated glomeruli are incubated in PCF-free media, the ferritin coat is shed form the free surfaces within 1/2 to 1 hour except at the tips and sides of microvillous projections. Pretreatment of normal glomeruli with neuraminidase or protamine sulfate results in a dramatic reduction in the binding of PCF to the glomerular epithelial free surface. Treatment of PAN glomeruli with PCF often results in patchlike distributions of PCF over the glomerular epithelial free surface. High concentrations of PCF (1.0 mg/ml) bind preferentially to microvillous projections on the parietal epithelial luminal surface and in distinct patchlike patterns along the basal laminae of parietal and tubular epithelial cells.