IMMUNOCYTOCHEMICAL STUDY OF GLOMERULAR-PERMEABILITY TO ANIONIC, NEUTRAL AND CATIONIC ALBUMINS

The renal handling of albumin of various isoelectric points (pI) was studied in mice by high resolution quantitative immunocytochemistry. Bovine serum albumin (BSA) was tagged with dinitrophenol (DNP) and cationized to pI 6.5 to 7.5 and to pI over 8.5. These tracers, including the anionic BSA-DNP (pI 4.8) were injected in the iliac vein of mice and the post-embedding protein A-gold technique was used with antibodies against DNP to localize the different tracers in renal tissue. Morphometric analysis of the labeling over the glomerular wall has demonstrated variations in its distribution according to the pI of the markers. Anionic and neutral BSA molecules were detected on the endothelial side of the basement membrane, while labeling for cationic BSA was present mainly in the laminae rarae interna and externa known to display fixed polyanionic sites. Neutral and cationic BSA-DNP reached the urinary space and were detected within the endocytotic apparatus of the proximal tubule epithelium. Neutralization of the anionic sites of the basement membrane with cationic BSA resulted in an increase in the permeability towards anionic BSA-DNP. In addition to the demonstration of glomerular permeability properties towards various probes, the present study has demonstrated that dinitrophenylated albumin represents an excellent versatile tool for the quantitative morphological investigation of glomerular permeability.