Spontaneous oscillations of intracellular calcium and growth hormone secretion.
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.
Holl
R W
RW
Department of Internal Medicine, University of Virginia, Charlottesville 22908.
Thorner
M O
MO
Mandell
G L
GL
Sullivan
J A
JA
Sinha
Y N
YN
Leong
D A
DA
D000818:Animals; D001572:Benzofurans; D002118:Calcium / Q000378:metabolism*; D003035:Cobalt / Q000494:pharmacology; D005089:Exocytosis; D005456:Fluorescent Dyes; D016257:Fura-2; D013006:Growth Hormone / Q000378:metabolism*; D013007:Growth Hormone-Releasing Hormone / Q000494:pharmacology; D006462:Hemolytic Plaque Technique; D007473:Ion Channels / Q000378:metabolism; D008297:Male; D008856:Microscopy, Fluorescence; D010903:Pituitary Gland, Anterior / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D013004:Somatostatin / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D001572:Benzofurans; D005456:Fluorescent Dyes; D007473:Ion Channels; D003035:Cobalt; D013004:Somatostatin; D013006:Growth Hormone; D013007:Growth Hormone-Releasing Hormone; D014700:Verapamil; D002118:Calcium; D016257:Fura-2