In:
American Journal of Physiology-Renal Physiology, American Physiological Society, Vol. 279, No. 1 ( 2000-07-01), p. F65-F76
Abstract:
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 β-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 ( V rev , in mV) near equilibrium potential for Cl − ( E Cl ) ( V rev = −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 V rev at E K ( V rev = −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 K v LQT 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.
Type of Medium:
Online Resource
ISSN:
1931-857X
,
1522-1466
DOI:
10.1152/ajprenal.2000.279.1.F65
Language:
English
Publisher:
American Physiological Society
Publication Date:
2000
detail.hit.zdb_id:
1477287-5
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