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J. Biol. Chem., Vol. 266, Issue 35, 23739-23744, 12, 1991
E Gorczynska and DJ Handelsman
Sertoli cells are hormonally regulated by follicle-stimulating hormone
(FSH) acting upon a G-protein-linked cell surface FSH receptor. FSH
increases intracellular cyclic AMP but the involvement of other signal
transduction mechanisms including intracellular calcium in FSH action are
not proven. Using freshly isolated rat Sertoli cells we measured cytosolic
free ionized calcium levels by dual-wavelength fluorescence
spectrophotometry using the calcium-sensitive fluorescent dye Fura2-AM. The
cytosolic calcium concentration in unstimulated Sertoli cells was 89 +/- 2
nM (n = 151 experiments) and was markedly increased by either calcium
channel ionophores (ionomycin, Bay K8644) or plasma membrane depolarization
consistent with the presence of voltage-sensitive and - independent calcium
channel in Sertoli cell membranes. Ovine FSH stimulated a specific,
sensitive (ED50, 5.0 ng of S-16/ml), and dose- dependent (maximal at 20
ng/ml) rise in cytosolic calcium commencing within 60 s to reach levels of
192 +/- 31 nM after 180 s and lasting for at least 10 min. The effect of
FSH was replicated by forskolin, cholera toxin, and dibutyryl cyclic AMP,
suggesting that cyclic AMP may mediate the FSH-induced rise in cytosolic
calcium. The FSH-induced rise in cytosolic calcium required extracellular
calcium and was abolished by calcium channel blockers specific for
dihydropyridine (verapamil, nicardipine), nonvoltage-gated (ruthenium red)
or all calcium channels (cobalt). Thus FSH action on Sertoli cells involves
a specific, rapid, and sustained increase in cytosolic calcium which
requires extracellular calcium and involves both dihydropyridine-sensitive,
voltage-gated calcium channels and voltage-independent, receptor-gated
calcium channels in the plasma membranes of rat Sertoli cells. The
replication by cyclic AMP of the effects of FSH suggests that calcium may
be a signal-amplification or -modulating mechanism rather than an alternate
primary signal transduction system for FSH in Sertoli cells.
The role of calcium in follicle-stimulating hormone signal transduction in Sertoli cells
Department of Obstetrics and Gynecology, University of Sydney, New South Wales, Australia.
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