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J. Biol. Chem., Vol. 263, Issue 20, 9682-9685, 07, 1988
RW Holl, MO Thorner, GL Mandell, JA Sullivan, YN Sinha and DA Leong
A novel combination of two single cell assays allowed the simultaneous
measurement of intracellular calcium concentration and hormone secretion in
normal pituitary cells. [Ca2+]i was recorded using the fluorescent Ca2+
indicator fura-2 and digital imaging microscopy. This technique was
combined with a reverse hemolytic plaque assay for growth hormone in order
to identify somatotropes and quantitate the amount of hormone released. A
dynamic profile of rhythmic calcium oscillations was found in spontaneously
secreting somatotropes. Each somatotrope displayed a distinct frequency
(one pulse every 5-30 s) and amplitude (range 50-450 nM) generated
asynchronously from cell to cell. The amount of growth hormone (GH)
released correlated directly with both the frequency and amplitude of
calcium oscillations at the level of single GH cells. Furthermore, calcium
excursions in somatotropes were rapidly suppressed by either (i) removal of
extracellular calcium, (ii) somatostatin (1 mM), or (iii) the calcium
channel blockers cobalt (2 mM) and verapamil (100 microM). These
observations demonstrate that spontaneous calcium oscillations are
characteristic for normal somatotropes. These oscillations are related to
spontaneous hormone secretion and due to influx through calcium channels in
the membrane. Somatostatin, the physiologic inhibitor of GH secretion,
suppresses calcium transients. These findings suggest that the
intracellular signaling information may be encoded both in the frequency
and amplitude of calcium oscillations.
Spontaneous oscillations of intracellular calcium and growth hormone secretion
Department of Internal Medicine, University of Virginia, Charlottesville 22908.
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