Cytosolic Calcium Transient and Light Response in Limulus Ventral Nerve Photoreceptors: Supralinearity in Response-Intensity Curves

Abstract

Abstract Light-evoked intracellular electrical responses were measured simultaneously with the changes in intracellular free Ca2+ or Sr2+ concentration by means of arsenazo III. The response-versus-stimulus intensity characteristics was measured in two states of relative light adaptation in strontium-containing saline (10 mᴍ Sr2+; 0.25 mᴍ Ca2+). Supralinear parts of the curves with slopes > 1 were found for the receptor current amplitude and the current­-time integral as well as for the amplitude of the arsenazo signal within at least 0.3 log units of identical stimulus intensity range. Maximum slopes of response-versus-intensity curves are on average approximately 1.7 for the receptor current and the arsenazo signal in both adaptational states. The rise in time of the arsenazo signal evoked by a light flash shows two phases. In strontium saline we observe a fast increase at the onset of the arsenazo signal which is detectable within the first 20-100 ms of the signal rise and a slow further increase to the maximum, with a duration of 0.7-6 s. In the detailed analysis of both phases we observed that the amplitude of each phase increases supralinear, i.e. disproportionate to the stimulus intensity. The fast rising phase of the arsenazo signal is not visibly voltage-dependent. This fast rising phase might be a monitor of an IP3-sensitive calcium or strontium release. Since the amplitude of the fast rising phase increases supralinear with the stimulus intensity, we propose the interpretation that more than one IP3-molecule must bind to the IP3-receptor to open this channel of the intracellular calcium store. Only the slow rising phase is voltage-dependent. The kinetic of this slow rising phase was accelerated with more negative membrane potentials. This predominant slow phase correlates with the receptor current-time integral. Therefore the slow rising phase of the arsenazo signal might be a monitor for influx of calcium or strontium through light activated ion channels in the plasma membrane. The supralinearity of the amplitude of the slow rising phase might indicate the supralinear increase in calcium or strontium, which is carried by the receptor current. Additionally, we calculated the fraction of strontium and calcium ions of the total receptor current. When calcium was replaced by strontium in the superfusate about 3-30% of the receptor current are carried by strontium and calcium ions. In physiological saline calcium contributes about 1-1.5% to the receptor current.