Calcium efflux from the rat neurohypophysis.

1. Calcium efflux from isolated rat neurophypophyses has been studied. Curve fitting of the wash-out curves suggests three phases with t((1/2)) of ca. 3, 15 and 130 min.2. The slow component of the (45)Ca efflux is attributed to efflux of intracellular Ca. On the basis of the temperature sensitivity of the Ca efflux, the activation energy has been calculated to be approximately 12,000 cal/mole, corresponding to a Q(10) of ca. 2.0.3. Ca efflux decreased by approximately 32% when external Na was replaced by choline. Li(o), in the presence or absence of Ca(o), was as effective as Na(o) in stimulating the Ca efflux.4. The curve relating Ca efflux to Na or Li is sigmoid and suggests that at least two Na (or Li) ions are necessary to activate the efflux of each Ca ion. Ca(o) does not modify the absolute Na-dependent Ca efflux but decreases the affinity for Na of the site involved in Ca extrusion.5. Removal of Ca(o) decreased the Ca efflux by ca. 44% in Na-free media. The apparent affinity for Ca(o) of the Ca(o)-activated Ca efflux (K(m) (Cao) = 20 muM) is greatly decreased by the presence of 150 mM-Na (K(m) (Cao) = 0.8 mM).6. Lanthanum decreased the total Ca efflux by ca. 60% and totally abolished the Na(o)-activated and Ca(o)-activated Ca efflux.7. Vanadate reduced the Ca efflux remaining in Na-, Ca-free saline by 73%.8. Elevation of Na(i) with ouabain did not modify the rate of loss of (45)Ca.9. Increased concentration of K(o) stimulated transiently the (45)Ca loss. The time course of this increase depends on the Ca(o) concentration (Ca).10. Cyanide or CCCP (carbonyl cyanide m-chlorophenylhydrazone) increased transiently the Ca efflux. The increase induced by cyanide could only be observed when the neural lobes had been over-loaded with (45)Ca.11. Membrane destruction induced by high temperature eliminated the effect of Na and Ca on (45)Ca efflux.12. In 150 mM-Na-containing saline, half-maximum activation of (45)Ca uptake occurs in the 0.2-0.4 mM Ca range.13. The Ca efflux from isolated pituicytes was not affected by removal of Na(o).14. In conclusion we show that Ca efflux from neurosecretory nerve terminals can be subdivided into three components of approximately the same magnitude, one which is activated by Na(o), another by Ca(o) and a third component which is independent of Na(o) and Ca(o).