AJP - Renal Physiology, Vol 251, Issue 2 214-F219, Copyright © 1986 by American Physiological Society
Use of trypan blue for identifying early proximal convoluted tubules
M. A. Linshaw, C. A. Bauman and L. W. Welling
Proximal convoluted renal tubules (PCT) are difficult to dissect for in
vitro study, particularly in newborn animals, and are identified with
certainty only when obtained in continuity with their parent glomeruli. We
show here that the supravital colloidal dye trypan blue preferentially
stains early PCT segments of rabbit in vivo and greatly facilitates their
isolation. We also show that trypan blue administration does not adversely
affect animal growth rate, kidney weight, or several general measures of
kidney function. Using isolated PCT from newborn to adult rabbits, control
(unstained) and trypan blue-stained tubules also are found not to differ in
lateral membrane surface density, in their ability to maintain normal cell
volume or to respond to hypotonic osmotic challenge, and in their swelling
rate in isotonic medium after ouabain treatment. We conclude that trypan
blue has no adverse effects on PCT cell function while greatly facilitating
their isolation for physiological studies.
