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J. Biol. Chem., Vol. 259, Issue 11, 7129-7136, 06, 1984
MA Beaven, J Rogers, JP Moore, TR Hesketh, GA Smith and JC Metcalfe
Rat basophil leukemic (2H3) cells ( Siraganian , R.P., McGivney , A.,
Barsumian , E. L., Crews, F. T., Hirata , F., and Axelrod , J. (1982) Fed.
Proc. 41, 30-34) loaded with fluorescent Ca2+ indicator quin 2 ( Tsien , R.
Y. (1980) Biochemistry 19, 2396-2404) showed a rapid increase in free
cytosol calcium concentration [( Ca]i) when histamine release was induced.
Intracellular quin 2 concentrations up to 7 mM did not affect release of
histamine in response to antigen (aggregated ovalbumin) or concanavalin A
with cells primed with antigen-specific monoclonal IgE, or in response to
Ca2+ ionophores. The [Ca]i increased from approximately 105 nM to a maximum
of approximately 1200 nM within 2 to 3 min after antigenic stimulation and
then declined slowly over 30 min toward the level in unstimulated cells.
Histamine release was most rapid as [Ca]i reached the maximum value and
then decreased continuously with [Ca]i over the subsequent 30 min. Neither
the Ca signal nor histamine release was observed when the Ca2+
concentration in the medium [( Ca]o) was less than 50 microM, but both
responses were restored on readdition of Ca2+ to 1 mM. The maximal Ca
signal was obtained when [Ca]o was approximately greater than 1 mM and was
half- maximal at [Ca]o congruent to 0.4 mM. In marked contrast [Ca]i in
unstimulated cells varied very little with [Ca]o from 0.1 to 1 mM.
Maintenance of the Ca signal required the continuous presence of
stimulating ligand, external Ca2+, and the maintenance of cellular ATP;
metabolic inhibitors blocked or reversed the Ca signal. La+ ions also
caused a rapid and reversible block of the Ca signal and histamine release.
The data are interpreted in a model in which the Ca signal is generated by
a La3+-sensitive signal influx pathway that is functionally independent of
the normal Ca2+ influx pathway in unstimulated cells, and that allows a
10-fold or greater increase in rate of Ca2+ entry. The Ca signal is
maintained dynamically by the balance between the increased Ca2+ influx and
active Ca2+ efflux across the plasma membrane.
The mechanism of the calcium signal and correlation with histamine release in 2H3 cells
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