Electrogenic Na(+)-Ca2+ exchanger, the link between intra- and extracellular calcium in the Limulus ventral...

1. Limulus ventral photoreceptors were injected with Arsenazo III and the internal change in the calcium concentration, [Ca2+]i, was measured under voltage clamp conditions. It is shown that in response to a light flash the rising phase of the [Ca2+]i is independent of the clamp voltage, Vm. This observation is contrary to other results reported in the literature. Experiments are reported that resolve this contradiction (see paragraph 4). 2. The relaxation of the [Ca2+]i after a bright light flash was observed to have a fast and slow phase. A function consisting of the sum of an exponential and a ramp was fitted to the relaxation. The fast phase, characterized by the time constant of the exponential, was observed not to depend on Vm, while the slow phase, characterized by the slope of the ramp, was strongly dependent on Vm. Furthermore the slope of the slow phase is shown to depend on the external Na+ concentration, but not the time constant of the fast phase. 3. In the dark the [Ca2+]i was observed to increase when the cell was depolarized and to decrease when the cell was hyperpolarized. This observation was more pronounced when the cell was continuously illuminated. 4. When the cell was clamped to a depolarizing voltage before illumination of the cell, the maximum of the calcium indicator signal was observed to depend on how long the cell had been clamped before applying the light stimulus. This experiment resolves the contradiction mentioned in paragraph 1. 5. The results presented here are consistent with the interpretation that a Na(+)-Ca2+ exchanger with a stoichiometry greater than 2:1 is the predominant link between intra- and extracellular calcium. Secondly that the light-induced intracellular calcium increase comes from a release by intracellular stores. Finally a measurable uptake of calcium occurs after a light-induced release, possibly by the internal calcium stores. The two-phase recovery of [Ca2+]i after a light flash is interpreted as being a calcium uptake by the internal stores, the fast phase, and removal by the electrogenic Na(+)-Ca2+ exchanger, the slow phase.