Interaction between cationic dyes and erythrocyte membranes in minimal change nephropathy: an electrophoretic approach.
A study was undertaken to clarify the usefulness of two cationic dyes, alcian blue (AB) and ruthenium red (RR) in demonstrating the defect in cellular membranes noted in minimal change nephropathy (MCN). The binding of both dyes to RBC membranes purified from normal and nephrotic children was evaluated by electrophoretic titration curves. When examined separately, AB was found to precipitate spontaneously, producing macro-aggregates with no electrophoretic mobility at pH 5. This was presumed to be the result of hydrophobic interaction of the dye with itself. The same phenomenon was observed when this dye was incubated at 37 degrees C with RBC ghost's from normal children, when AB presented a sigmoidal curve with a net positive charge for pHs higher than 5.5 and lower than 5 and no electrophoretic mobility at pH 5. However, incubation of AB with RBC ghosts from children with MCN resulted in an improvement of the solubility of the dye which then migrated with a net positive charge along the whole gradient of pH from 3.5 to 9. The presence of zwitterionic neutral detergents such as CHAPS, but not of a charged substance such as protamine sulphate, inhibited precipitation at pH 5 when incubated with membranes from normal children, supporting the hydrophobic nature of the phenomenon. When RR was used instead of AB, it was fully protonated (i.e. did not precipitate) when analysed alone, but when incubated with normal RBC ghosts, it also revealed no electrophoretic mobility at pH 5.(ABSTRACT TRUNCATED AT 250 WORDS)
Candiano
G
G
Nephrology Department, G. Gaslini Institute, Genoa, Italy.
Ghiggeri
G M
GM
Oleggini
R
R
Ginevri
F
F
Altieri
P
P
Gusmano
R
R
D000423:Alcian Blue / Q000378:metabolism*; D002648:Child; D002675:Child, Preschool; D004586:Electrophoresis; D004910:Erythrocyte Membrane / Q000378:metabolism*; D006801:Humans; D007678:Kidney Glomerulus / Q000378:metabolism; D009402:Nephrosis, Lipoid / Q000378:metabolism*; D012430:Ruthenium Red / Q000378:metabolism*
D012430:Ruthenium Red; D000423:Alcian Blue