Stimulation by aldosterone of the sodium efflux in barnacle muscle fibres: effects of RNA inhibitors and spironolactone

Abstract

1. Single muscle fibres from the barnacle Balanus nubilus have been studied to provide information about the mode of action of aldosterone on Na transport in a symmetric cell. 2. Neither internal nor external application of aldosterone modified the course of the Na efflux. However, fibres pre-exposed overnight to a dose of aldosterone showed a biphasic response to external aldosterone. The first phase was prompt in onset and consisted of a falling rate constant for Na efflux becoming a constant. This has been interpreted as indicating that aldosterone acts by releasing sequestered or bound Na. The second phase was delayed in onset with the average latent period being 68 min. It consisted of a step-up in the rate of Na efflux, followed by a transitory rise in the efflux rate constant. This second phase was dose-dependent, the minimum concentration being 10(-9) M. 3. Internal application of aldosterone in a concentration as low as 10(-10) M promptly stopped the rate constant for Na efflux from further falling but failed to cause delayed stimulation. 4. The response of the Na efflux into Li-ASW following external application of aldosterone was bi-phasic. The magnitude of the delayed stimulation was comparable to that found in controls. 5. No correlation between latent period or size of the internal Na bound fraction, and the magnitude of the delayed stimulation could be established. 6. Internal application of actinomycin-D, alpha-amanitin or cordycepin shortly before application of external aldosterone caused complete abolition of the delayed stimulation. 7. Overnight pre-exposure of the barnacle to actinomycin-D caused complete abolition of the falling rate constant for Na efflux, as well as the delayed stimulation caused by external aldosterone. 8. Internal application of spironolactone SC-14266 shortly before external application of aldosterone caused complete abolition of the biphasic response to the steroid. 9. It is concluded that (i) barnacle fibres can be made sensitive to aldosterone, (ii) the biphasic effects on the Na efflux depend on prior induction of RNA, (iii) the first effect caused by internal or external aldosterone involves mobilization of sequestered Na, (iv) the results obtained with spironolactone are in keeping with the current view that aldosterone interacts with a receptor system before causing de-repression. 10. The implications of the finding that aldosterone releases sequestered Na are briefly touched upon.