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Volume 271, Number 49, Issue of December 6, 1996 pp. 31210-31214
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Differential Regulation of Nuclear and Cytosolic Ca²⁺ in HeLa Cells

(Received for publication, June 17, 1996)

From the Department of Medical Biochemistry, University of Wales College of Medicine, Heath Park, Cardiff, CF4 4XN, United Kingdom and the ^§ Cardiovascular Division, University of Virginia Health Sciences Center, Charlottesville, Virginia 22908

The results reported in this study address the controversial issue that nuclear free Ca²⁺ ([Ca²⁺]_n) may be regulated independently of cytosolic free Ca²⁺ ([Ca²⁺]_c). We have measured [Ca²⁺]_n and [Ca²⁺]_c with recombinant aequorin targeted to the nucleus and cytosol in HeLa cells. We found that histamine, ATP, and ionomycin increased [Ca²⁺]_c quantitatively more than [Ca²⁺]_n, although the time course of these changes was similar. The difference between [Ca²⁺]_c and [Ca²⁺]_n depended on the stimulus, and the relative difference between [Ca²⁺]_n and [Ca²⁺]_c was less with ionomycin than with histamine or ATP. After depletion of the internal Ca²⁺ store, restoration of extracellular Ca²⁺ resulted in only increased [Ca²⁺]_c without a significant increase in [Ca²⁺]_n. Treatment with cyclopiazonic acid resulted in a delayed increases in [Ca²⁺]_n compared to [Ca²⁺]_c. These differences in both timing and magnitude of nuclear Ca²⁺ signals confirm that the cell can limit or delay increases in nuclear free Ca²⁺. Taken with the fact that an inositol phosphate signaling system resides in the nucleus and its envelope, our data support the hypothesis that [Ca²⁺]_n may be independently regulated.

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