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Science 16 February 1990:
Vol. 247. no. 4944, pp. 858 - 862
DOI: 10.1126/science.2154851
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Articles

Science, Vol 247, Issue 4944, 858-862
Copyright © 1990 by American Association for the Advancement of Science

articles

Subcellular calcium transients visualized by confocal microscopy in a voltage-clamped vertebrate neuron

A Hernandez-Cruz, F Sala, and PR Adams

Howard Hughes Medical Institute, Department of Neurobiology and Behavior, State University of New York, Stony Brook 11794.

Confocal laser-scanned microscopy and long-wavelength calcium (Ca2+) indicators were combined to monitor both sustained and rapidly dissipating Ca2+ gradients in voltage-clamped sympathetic neurons isolated from the bullfrog. After a brief activation of voltage-dependent Ca2+ channels, Ca2+ spreads inwardly, and reaches the center of these spherical cells in about 300 milliseconds. Although the Ca2+ redistribution in the bulk of the cytosol could be accounted for with a radial diffusion model, local nonlinearities, suggesting either nonuniform Ca2+ entry or spatial buffering, could be seen. After electrical stimulation, Ca2+ signals in the nucleus were consistently larger and decayed more slowly than those in the cytosol. A similar behavior was observed when release of intracellular Ca2+ was induced by caffeine, suggesting that in both cases large responses originate from Ca2+ release sites near or within the nucleus. These results are consistent with an amplification mechanism involving Ca2(+)-induced Ca2+ release, which could be relevant to activity-dependent, Ca2(+)-regulated nuclear events.


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