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1 Physiologisches Institut and 2 Medizinische Poliklinik der Ludwig-Maximilians-Universität, 80336 Munich; and 3 Institut für Anatomie der Charité, 10098 Berlin, Germany
The expression
patterns of plasma membrane transporters that specify the
epithelial cell type are acquired with ontogeny. To study this process
during metanephrogenic mesenchyme-to-epithelium transition, branching
ureteric buds with their adjacent mesenchymal blastema (mouse embryonic
day E14) were dissected and explanted on a collagen matrix.
In culture, induced mesenchymal cells condensed, aggregated, and
converted to the comma- and S-shaped body. During in vitro condensation
and aggregation, transcription factor Pax-2 protein was downregulated
while the epithelial markers E-cadherin and 
-catenin proteins were
upregulated. In addition, Wilms' tumor suppressor protein WT-1 was
detectable upon condensation and downregulated in the S stage, where
expression persisted in the long arm of the S. Patch-clamp, whole cell
conductance (G, in nS/10 pF) of pre-epithelial condensed
mesenchymal cells (n = 7) was compared with that of
tubular proximal S-shaped-body epithelium (n = 6). Both
stages expressed E-cadherin and WT-1 mRNA, as demonstrated by
single-cell RT-PCR, testifying further to the epithelial as well as the
nephrogenic commitment of the recorded cells. Mesenchymal cells
exhibited whole cell currents (G = 6.7 ± 1.3)
with reversal potentials (Vrev, in mV) near
equilibrium potential for Cl
(ECl)
(Vrev = 
40 ± 7) suggestive of a
high fractional Cl conductance. Currents of the
S-shaped-body cells (G = 4.0 ± 1.1), in sharp
contrast, had a Vrev at
EK (Vrev = 82 ± 6) indicating a high fractional K+ conductance. Further,
analysis of K+-selective whole cell tail currents and
single-channel recording revealed a change in K+ channel
expression. Also, Kir6.1 K+ channel mRNA and protein were
downregulated between both stages, whereas KvLQT
K+ channel mRNA was abundant throughout. In conclusion,
metanephrogenic mesenchyme-to-epithelium transition is accompanied by a
profound reorganization of plasma membrane ion channel conductance.
mesenchymal-epithelial transition; embryonic ion channels; single-cell reverse transcription-polymerase chain reaction; kidney developmental biology
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