Role of de novo protein synthesis and calmodulin in rapid activation of Na(+)-H+ exchange of aldosterone in frog diluting segment.

AUTOR(ES)

Cooper, G J

RESUMO

1. In the amphibian early distal tubule aldosterone activates the Na(+)-H+ exchangers, resulting in an increase in intracellular pH (pHi). Since this activation is rapid (within 30 min), it may be mediated by either a genomic or non-genomic pathway. 2. pHi was measured in single microperfused early distal tubule segments using the fluorescent probe 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF). 3. A 30 min incubation in aldosterone increased both resting pHi and the setpoint of the Na(+-H+ exchanger. These changes were prevented by the mineralocorticoid receptor antagonist, spironolactone. 4. Actinomycin D and cycloheximide, inhibitors of transcription and translation, respectively, were without effect on resting pHi, but inhibited activation of the Na(+)-H+ exchanger by aldosterone. 5. The effect of aldosterone upon pHi and setpoint was also prevented by the calcium-calmodulin antagonist, W-7. 6. These results indicate that, although the response to aldosterone is rapid, aldosterone binds to a specific mineralocorticoid receptor which then triggers gene activation followed by de novo protein synthesis. Furthermore, since calmodulin is a known activator of the Na(+)-H+ exchanger, and the response is inhibited by W-7, it is suggested that this protein may be calmodulin.

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