Calcium efflux from Myxicola giant axons: effects of extracellular calcium and intracellular EGTA

Abstract

1. (45)Ca efflux was examined in Myxicola giant axons injected with (45)CaCl(2) or various concentrations of (45)Ca/EGTA buffers. In axons injected with (45)CaCl(2), the Ca(o)-dependent Ca efflux in 1 mM-Ca(o) was about half that in 10 mM-Ca(o).2. Axons injected with (45)Ca/EGTA buffers consistently showed two types of results: in one type (B type), K((1/2)) for Ca(o) activation was less than 1 mM-Ca(o). In the other type of result (A type), there was an additional Ca activation of Ca efflux. This additional efflux exhibited a linear dependence on Ca(o) when the Ca(o) values ranged between 1 mM-Ca(o) and 10 mM-Ca(o).3. The B-type result remained unchanged when the injected Ca/EGTA concentrations were varied. The A type result, however, changed as a function of Ca/EGTA(i) in the following way: (a) at a constant ratio of Ca/EGTA(i) = 8/10, the megnitude of the linear component of the Ca(o)-activated Ca efflux was reduced by increasing the intracellular concentration of (Ca/EGTA) buffer; and (b) at a ratio Ca/EGTA = 1/10, the linear component of the Ca(o)-activated Ca efflux appeared to acquire a slower time response to changes in Ca(o).4. Na(o) acts synergistically with Ca(o) to produce the linear component of the Ca-activated Ca efflux seen in the A type result.5. With axons containing (45)Ca/EGTA buffers (total EGTA(i) = 1 mM), changing the Ca/EGTA ratio by repetitive injections of CaCl(2) did not increase (45)Ca efflux by as great an amount as would be predicted if Ca(i) (2+) were controlled by the EGTA buffer alone.6. Ca(i) (2+) (measured by arsenazo III absorbance) is influenced by Ca(o) irrespective of the presence of 1 mM-EGTA buffer inside the axon. There was a variability in the sensitivity of Ca(i) to Ca(o) that resembled the variability found in (45)Ca efflux measurements.7. (45)Ca influx is not affected by the concentration of Ca/EGTA buffer injected into the cell and appears to be only slightly, if at all, affected by increasing ionized Ca(i) (2+) from 0.016 to 0.56 muM in the injection medium.8. These results are consistent with the interpretation that the Ca efflux system of the Myxicola giant axon, or something controlling it, can exist in more than one state. One of these states, which exhibited a large Ca(o)-dependent Ca efflux, may represent axons in which Ca(i) is poorly controlled by the natural endogenous Ca buffers.