Intracellular Ca2+ dynamics during spontaneous and evoked activity of leech heart interneurons: low-threshold Ca...

In oscillatory neuronal networks that pace rhythmic behavior, Ca(2+) entry through voltage-gated Ca channels often supports bursting activity and mediates graded transmitter release. We monitored simultaneously membrane potential and/or ionic currents and changes of Ca fluorescence (using the fluorescence indicator Ca Orange) in spontaneously active and experimentally manipulated oscillator heart interneurons in the leech. We show that changes in Ca fluorescence in these interneurons during spontaneous bursting and evoked activity reflect the slow wave of that activity and that these changes in Ca fluorescence are mediated by Ca(2+) entry primarily through low-threshold Ca channels. Spatial and temporal maps of changes in Ca fluorescence indicate that these channels are widely distributed over the neuritic tree of these neurons. We establish a correlation between the amount of transmitter released, as estimated by the integral of the postsynaptic current, and the change in Ca fluorescence. In experiments in which we were able to record presynaptic low-threshold Ca currents, associated IPSCs, and presynaptic changes in Ca fluorescence from fine neuritic branches of heart interneurons near their region of synaptic contact with their contralateral partner, there was a close association between the rise in Ca fluorescence and the rise of the postsynaptic conductance. The changes in Ca fluorescence that we record at the end of fine neuritic branches appear to reflect changes in [Ca(2+)](i) that mediate graded synaptic release in leech heart interneurons. These results indicate that widely distributed low-threshold Ca currents play an important role in generating rhythmic activity and in mediating graded transmitter release.