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Mechanisms of Aldosterone Action on NCC

Aldosterone increases NCC function and protein expression in whole animal studies at 72 hours but more acute effects of aldosterone on NCC activity have not been studied. We have recently demonstrated that in a mammalian cell model with robust native NCC activity (mDCT15), aldosterone stimulated NCC activity by 26% at 6 hrs and by 52% at 24 hrs. However, the mechanism by which this acute stimulation was mediated is unknown. To examine this question, mDCT15 cells were incubated with either 100nM aldosterone or vehicle (DMSO) for 6 or 24 hours. Cells were then surface biotinylated and surface expression was assessed by Western blotting. Total NCC protein expression remained unchanged compared to control in both the 6 and 24 hour treatment groups. Furthermore, NCC surface expression did not change in the 6 or 24 hour aldosterone treated groups compared to control. This suggests that the effects of aldosterone upon NCC are not primarily mediated by increased surface expression, but instead due to an increase in individual transporter activity. Further work is needed to characterize this mechanism.