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Acute Effects of Adenosine Triphosphates, Cyclic 3',5'-Adenosine Monophosphates, and Follicle-Stimulating Hormone on Cytosolic Calcium Level in Cultured Immature Rat Sertoli Cells

^a Laboratoire de Physiologie des Régulations Cellulaires, UMR 6558, UFR Sciences, 86022-Poitiers-Cedex, France

The ability of ATP and FSH to induce intracellular calcium [Ca²⁺]_i changes in Sertoli cells is imperfectly understood and reports are conflicting. We have applied the single-cell microfluorometry technique with the calcium probe indo-1 to investigate [Ca²⁺]_i in individual cultured Sertoli cells. When cells were exposed to ATP, cAMP, and FSH, a fast and biphasic increase in [Ca²⁺]_i was obtained in 100%, 70%, and 56% of cells, respectively. Caffeine did not activate Ca²⁺ mobilization, while thapsigargin suppressed the peak response. External calcium free-EGTA buffer suppressed the plateau phase, while blockers of voltage-operated Ca²⁺ channels did not abolish the response to cAMP and ATP. We conclude that the three messengers mobilized Ca²⁺ from intracellular thapsigargin-sensitive stores, which induced a subsequent Ca²⁺ influx from the extracellular medium by a voltage-independent Ca²⁺ entry. The well-documented mechanisms by which these messengers act on cells support the idea that they release Ca²⁺ from smooth endoplasmic reticulum by two different pathways, or that FSH and cAMP first release ATP, which then acts on cells. Among the cells, 77% and 80% responded, respectively, to FSH and cAMP by a delayed long-lasting decrease in [Ca²⁺]_i that was never recorded in the presence of ATP. This suggests that FSH and cAMP also promote a slow redistribution of [Ca²⁺]_i from the exchangeable pool to the bound nonexchangeable pools. Involvement of voltage-operated and voltage-independent calcium channels in the response of Sertoli cells to ATP, FSH, and cAMP is discussed.

¹ Correspondence: Michel Joffre, Laboratoire de Physiologie des Régulations Cellulaires, UMR 6558, UFR Sciences, 40 Avenue du Recteur Pineau, 86022-Poitiers-Cedex, France. FAX: 33 5 49 45 40 14; m.joffre{at}campus.univ-poitiers.fr

² Current address: Nathalie Lalevée, IGH, UPR 1142 CNRS, 141 rue de la Cardonille, 34 396 Montpellier, Cedex 5, France.

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