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Department of Physiology, Saga Medical School, Japan.

1. Dynamic changes in the intracellular free Ca2+ concentration ([Ca2+]i) following electrical membrane activity, were recorded from the neurone soma of the excised bullfrog sympathetic ganglion, using Fura-2 fluorescence and compared with the accompanying Ca(2+)-dependent electrical membrane responses. 2. The resting [Ca2+]i was about 100 nM, a value little changed by penetration with an intracellular electrode. 3. A net rise in fluorescence at a wavelength of 340 nm (Ca2+ transient) induced by a single action potential in Ringer solution rose almost in parallel with the initial decay phase of a slow Ca(2+)-dependent after-hyperpolarization; decayed in parallel with the late phase; and increased in amplitude and duration in the presence of tetraethylammonium (20 mM). 4. A Ca2+ transient induced by repetitive action potentials was increased asymptotically in amplitude and progressively in duration by increasing the number of spikes, and was slower in time course than the associated Ca(2+)-dependent K+ current. 5. Scanning a single horizontal line across the cytoplasm with an ultraviolet argon ion laser (351 nm) and recording Indo-1 fluorescence with a confocal microscope demonstrated an inward spread of a rise in [Ca2+]i following a tetanus. 6. Both single spike- and tetanus-induced Ca2+ transients were abolished in a Ca(2+)-free solution, while single or repetitive transient rises in [Ca2+]i induced by caffeine (5-10 mM) were generated under the same conditions. 7. Ryanodine (10-50 microM) did not affect tetanus-induced Ca2+ transients, whereas it blocked completely the caffeine-induced oscillation of [Ca2+]i. 8. Ca2+ transients induced by a tetanus in Ringer solution were independent of the interval from the preceding tetanus. The amplitude of Ca2+ transients induced by a tetanus in the presence of caffeine (5 mM) was equal to, or greater than, that generated in Ringer solution in any of the phases of [Ca2+]i oscillation. 9. It is suggested that under the physiological conditions here, the induction of action potentials does not cause the release of Ca2+ in the cells of the freshly excised bullfrog sympathetic ganglion, and that Ca(2+)-buffering systems contribute not only to lowering a transient rise in [Ca2+]i but also to sustaining an increased [Ca2+]i after a large Ca2+ load into the cell.

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