Corticosteroid regulation of amiloride-sensitive sodium-channel subunit mRNA expression in mouse kidney

Corticosteroid control of distal nephron sodium handling, particularly thro ugh the amiloride-sensitive sodium channel (ENaC), has a key role in blood pressure regulation. The mechanisms regulating ENaC activity remain unclear . Despite the generation of useful mouse models of disorders of electrolyte balance and blood pressure, there has been little study of distal nephron sodium handling in this species. To investigate how corticosteroids regulat e ENaC activity we isolated cDNA for the three mouse ENaC subunits (alpha, beta and gamma), enabling their quantitation by competitive PCR and in situ hybridisation. Kidneys were analysed from mice 6 days after adrenalectomy or placement of osmotic mini-pumps delivering aldosterone (50 mu g/kg per d ay), dexamethasone (100 mu g/kg per day), spironolactone (20 mg/kg per day) or vehicle alone (controls). In controls, renal ENaC alpha mRNA exceeded b eta or gamma by approximately 1.75- to 28-fold. AU subunit mRNAs were expre ssed in renal cortex and outer medulla, where the pattern of expression was fully consistent with localisation in collecting duct, whereas the distrib ution in cortex suggested expression extended beyond the collecting duct in to adjacent distal tubule. Subunit mRNA expression decreased from cortex to outer medulla, with a gradual reduction in beta and gamma, and ENaC alpha decreased sharply (similar to 50%) across the outer medulla. Expression of ENaC beta and gamma (but not alpha) extended into inner medulla, suggesting the potential for inner medulla collecting duct cation channels in which a t least ENaC beta gamma participate. Aldosterone significantly increased EN aC subunit expression; the other treatments had little effect. Aldosterone caused a 1.9- res 3.5-fold increase in ENaC alpha (particularly marked in o uter medullary collecting duct), but changes for beta and gamma were minor and limited to the cortex. The results raise the possibility that medullary ENaC alpha upregulation by aldosterone will create more favourable subunit stoichiometry leading to a more substantial increase in ENaC activity. In cortex, such a mechanism is unlikely to have a major role.