Abstract
Ontogeny of apical-basolateral epithelial cell polarity was studied by a comparison of embryonic ureteric bud (UB) and mature cortical collecting duct (CCD) primary cultures. Patch-clamp techniques were applied to identify apical ion channels and to measure specific conductance (in nS/10 pF) of the apical membrane (ga) and of the whole cell (Gw). In UB, Gw (1.3 +/- 0.17; n = 32) and ga (1.4 +/- 0.15; n = 9) were not different, whereas they differed greatly in CCD (Gw: 0.79 +/- 0.07, n = 35; ga: 2.3 +/- 0.38, n = 9). In UB, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) (100 microM) similarly reduced ga and Gw by 0.66 +/- 0.21 (n = 9) and 0.48 +/- 0.07 (n = 7), respectively, whereas DIDS had no effect in CCD. Amiloride (1 mM) did not alter UB conductance, whereas, in CCD, ga was more reduced than Gw (0.75 +/- 0.18, n = 8, vs. 0.22 +/- 0.10, n = 7). In addition to this Na(+)-selective conductance, a nonselective cation (NSC) conductance was enriched sixfold in the apical membrane of CCD, possibly due to apical 9 +/- 1-pS (n = 7) NSC channels, whereas UB apical NSC conductance was low. Apical 16 +/- 2-pS (n = 6) K+ channels were identified in CCD but not in UB. The comparison of both developmental states suggests a differentiation of the apical membrane during UB to CCD ontogeny.
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