Strontium-induced repetitive calcium spikes in a unicellular green alga

The divalent cation Sr2+ induced repetitive transient spikes of the cytosolic Ca2+ activity [Ca2+]cy and parallel repetitive transient hyperpolarizations of the plasma membrane in the unicellular green alga Eremosphaera viridis. [Ca2+]cy measurements, membrane potential measurements, and cation analysis of the cells were used to elucidate the mechanism of Sr2+-induced [Ca2+]cy oscillations. Sr2+ was effectively and rapidly compartmentalized within the cell, probably into the vacuole. The [Ca2+]cy oscillations cause membrane potential oscillations, and not the reverse. The endoplasmic reticulum (ER) Ca2+-ATPase blockers 2,5-di-tert-butylhydroquinone and cyclopiazonic acid inhibited Sr2+-induced repetitive [Ca2+]cy spikes, whereas the compartmentalization of Sr2+ was not influenced. A repetitive Ca2+ release and Ca2+ re-uptake by the ER probably generated repetitive [Ca2+]cy spikes in E. viridis in the presence of Sr2+. The inhibitory effect of ruthenium red and ryanodine indicated that the Sr2+-induced Ca2+ release from the ER was mediated by a ryanodine/cyclic ADP-ribose type of Ca2+ channel. The blockage of Sr2+-induced repetitive [Ca2+]cy spikes by La3+ or Gd3+ indicated the necessity of a certain influx of divalent cations for sustained [Ca2+]cy oscillations. Based on these data we present a mathematical model that describes the baseline spiking [Ca2+]cy oscillations in E. viridis.

Electrochemical Potential Gradients of H+, K+, Ca2+, and Cl- across the Tonoplast of the Green Alga Eremosphaera Viridis.

Using ion-selective microelectrodes, we measured the activity of H+, K+, Ca2+, and Cl- and the electrical potential both in the vacuole and in the cytoplasm of the unicellular green alga Eremosphaera viridis to obtain comparable values of the named parameters from the same object under identical conditions. The cytosol had a pH of 7.3, and activities of the other ions were 130 mM K+, 160 nM Ca2+, and 2.2 mM Cl-. We observed only small and transient light-dependent changes of the cytosolic Ca2+ activity. The vacuolar K+ activity did not differ significantly from the cytosolic one. The Ca2+ activity inside the vacuole was approximately 200 [mu]M, the pH was 5.0, and the Cl- activity was 6.2 mM. The concentrations of K+, Ca2+, and Cl- in cell extracts were measured by induction-coupled plasma spectroscopy and anion chromatography. This confirmed the vacuolar activities for K+ and Cl- obtained with ion-selective microelectrodes and indicated that approximately 60% of the vacuolar Ca2+ was buffered. The tonoplast potential was vanishingly low ([less than or equal to][plus or minus]2 mV). There was no detectable electrochemical potential gradient for K+ across the tonoplast, but there was, however, an obvious electrochemical potential gradient for Cl- (-26 mV), indicating an active accumulation of Cl- inside the vacuole.