AJP - Renal Physiology, Vol 252, Issue 3 412-F422, Copyright © 1987 by American Physiological Society
Immunological segmentation of the rabbit distal, connecting, and collecting tubules
P. Poujeol, P. Ronco, M. Tauc, M. Geniteau, F. Chatelet, D. Sahali, A. Vandewalle and P. Verroust
To obtain monoclonal antibodies (MAB) specific for the different cell types
of distal and collecting tubules, BALB/c mice were immunized with cell
suspensions highly enriched in cells from the distal segments of the rabbit
nephron. Nine MAB were selected and cloned. Four groups could be identified
on the basis of double-labeling immunofluorescence (IF) on frozen kidney
sections and on microdissected tubules. In addition, binding specificity at
the cellular level was studied by immunoelectronmicroscopy (IEM) for
selected MAB. A single MAB (group 1) was specific for distal bright cells
and a subpopulation of cortical ascending limb cells. Six MAB (group 2)
reacted with connecting and collecting tubules. Five of these (group 2A)
had similar binding patterns and reacted identically with the two tubular
segments. The MAB studied by IEM was specific for connecting and principal
cells. One antibody (group 2B) reacted with only a fraction of the cells
associated with the connecting tubule (CNT), but with all cells of the
cortical collecting tubule (CCT). By IEM, this antibody was found to be
specific for intercalated cells in CNT and bound both principal and
intercalated cells of the CCT. Two MAB (group 3) reacted with antigen(s)
expressed by the various terminal segments of renal tubule. MAB of groups 1
and 2A, which define distal bright cells and connecting-principal cells
from the CNT-CCT, respectively, were used for cell fractionation
experiments. Heterogeneous rabbit cortical cells were first incubated with
the selected MAB. MAB-bearing renal cells were separated on plastic dishes
previously coated with an affinity-purified goat anti-mouse immunoglobulin.
Using these procedures it was possible to obtain highly purified
subpopulations of distal, bright, or connecting-principal cells.
