Electrically and chemically mediated increases in intracellular calcium in neuronal growth cones

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Abstract
In the present report we used the calcium indicator fura-2 to compare intracellular levels of free calcium in growth cones of isolated Helisoma neurons under a variety of experimental conditions. We tested whether 2 different signals that inhibit growth cone motility--action potentials and serotonin--changed calcium levels in growth cones. Electrical stimulation of the cell body caused a rise in calcium levels at the growth cone. After brief stimulation, calcium levels quickly recovered to normal values, whereas longer stimulation periods required longer recovery times. The application of serotonin to growth cones caused an increase in calcium levels that was selective for growth cones of neurons whose outgrowth was inhibited by serotonin, but not for neurons whose outgrowth was not affected. We also found that motile growth cones had higher free calcium levels than growth cones that had spontaneously stopped growing. Furthermore, the distribution of calcium in neurons that contained motile growth cones was heterogeneous; calcium levels were always higher in the growth cone than in the neurite or soma. These data indicate that calcium levels in growth cones vary in different states of outgrowth and that calcium levels can be modulated by both electrical and chemical signals.