Thyroid hormone: Aldosterone antagonism in cultured epithelial cells

Abstract

Thyroid hormone (T 3) has been demonstrated to inhibit the action of aldosterone on sodium transport in toad urinary bladder and rat kidney. We have exammined the effect of T 3 on aldosterone action and specific nuclear binding in cultured epithelial cells derived from toad urinary bladder. In cell line TB6-C, addition of 5·10 −8 M T 3 to culture media for up to 3 days results in no change in short-circuit current or transepithelial resistance. This concentration of T 3 completely inhibits the maximal increase in short-circuit current in response to 1·10 −7 M aldosterone. The inhibition can be demonstrated with 18 h preincubation or with simultaneous addition of T 3 and aldosterone. The half-maximal concentration for the inhibition of the aldosterone effect is approx. 5·10 −9 M T 3. T 3 has no effect on cyclic AMP-stimulated short-circuit current in these cells. The effect of T 3 on nuclear binding of [ 3H]aldosterone was examined using a filtration assay with data analysis by at least-squares curve-fitting program. Best fit was obtained with a model for two binding sites. The dissociation constants for the binding were K′ d1 = (0.82 ± 0.36)·10 −10 M and K′ d2 = (3.2±0.60)·10 −8 M.The half-maximal concentration for aldosterone-stimulated sodium transport in these cells is approx. 1·10 −8 M. Analysis of nuclear aldosterone binding in cells preincubated for 18 h with 5·10 −8 M T 3 showed a K′ d1 = (0.15 ± 0.10)·10 −10 M and K′ d2 = (3.5 ± 0.10)·10 −8 M. We conclude that T 3 i action of aldosterone on sodium transport at a site after receptor binding in the nucleus.