Spironolactone antagonism of aldosterone action on Na+ transport and RNA metabolism in toad bladder epithelium
- 1 December 1977
- journal article
- research article
- Published by Springer Nature in The Journal of Membrane Biology
- Vol. 32 (1), 177-194
- https://doi.org/10.1007/bf01905216
Abstract
Summary In earlier studies, aldosterone increased the incorporation of precursors into a class of cytoplasmic RNA with the characteristics of messenger RNA (mRNA), in toad bladder epithelium. In the present studies, this effect was analyzed further with a competitive antagonist, spironolactone (SC-9420). Paired hemibladders were labeled with3H-uridine (30 min pulse–140 min chase), with or without aldosterone (3.5×10−8 m, 7×10−8 m) in the presence or absence of SC-9420 (7×10−6 m, 2.5×10−5 m) at molar ratios of 200∶1 to 280∶1. Cytoplasmic RNA, either the total phenol-SDS extract or polyadenylated-RNA (poly(A)(+)-RNA) obtained by oligo-deoxythymidylate-cellulose (oligo(dT)-cellulose) chromatography was analyzed in linear 5–20% sucrose gradients. Eight sets of experiments were completed in which the short-circuit current (scc) was monitored for 180 min and the incorporation of3H-uridine (30 min pulse–150 min chase) was simultaneously determined on pools of epithelia from 5 to 10 hemibladders. The fractional change inscc correlated linearly with the fractional change in3H-uridine of 12S cytoplasmic RNA (r=0.95,p3H-uridine into poly(A)(+)-RNA (particularly 12S). These results support the inference that induction of mRNA mediates the action of aldosterone on Na+ transport.Keywords
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