To study the role of extracellular nucleotides in the regulation of Sertoli cells, the effects of ATP and its analogs on the Ca2+-phospholipid- and cAMP-dependent pathways were tested. Cultured Sertoli cells from immature animals were incubated with ATP or structurally related compounds, and phosphoinositide (PI) turnover or cAMP accumulation was measured. Among the several nucleotide phosphate analogs tested, adenosine 5'-O-(3-thiotriphosphate) was the agonist most potent in stimulating inositol phosphate accumulation. The effects of purine nucleotides on PI turnover were time and concentration dependent. Because nonhydrolizable ATP analogs also stimulated PI turnover, ATP metabolites or metabolic products are not responsible for the observed stimulation. The order of potency of the different ATP analogs [adenosine 5'-O-(3-thiotriphosphate) > ATP congruent to UTP > beta,gamma-methyleneadenosine 5'-triphosphate, 2-methylthio-ATP > adenosine] was consistent with the presence of P-2U receptors (nucleotide receptors) on the surface of the Sertoli cell. Augmented PI turnover was accompanied by a transient increase in Ca2+ concentration, measured in single Sertoli cells loaded with the intracellular Ca2+ indicator fura-2. When used alone, ATP and its analogs did not have a direct effect on cAMP levels in the Sertoli cell. However, ATP or its analogs inhibited FSH-dependent cAMP accumulation by more than 70%. Purine nucleotides also efficiently blocked the effects of FSH distal to cAMP accumulation, because extracellular ATP completely reversed the changes in Sertoli cell shape induced by FSH. The nucleotide-dependent inhibition of cAMP accumulation was blocked by pertussis toxin to a different degree depending on the purine or pirimidine nucleotide used. This indicated that more than one mechanism contributes to the purine nucleotide-dependent inhibition of cAMP accumulation. These data provide evidence that purine nucleotide receptors coupled to multiple pathways are present on the Sertoli cell in culture, and that extracellular ATP has profound biological effects on the FSH responsiveness of the Sertoli cell.

ACTIVATION OF INOSITOL PHOSPHOLIPID TURNOVER AND CALCIUM SIGNALING IN RAT SERTOLI CELLS BY P2-PURINERGIC RECEPTORS - MODULATION OF FOLLICLE-STIMULATING-HORMONE RESPONSES / Filippini, Antonio; Riccioli, Anna; P., De Cesaris; R., Paniccia; A., Teti; Stefanini, Mario; M., Conti; Ziparo, Elio. - In: ENDOCRINOLOGY. - ISSN 0013-7227. - 134:3(1994), pp. 1537-1545. [10.1210/en.134.3.1537]

ACTIVATION OF INOSITOL PHOSPHOLIPID TURNOVER AND CALCIUM SIGNALING IN RAT SERTOLI CELLS BY P2-PURINERGIC RECEPTORS - MODULATION OF FOLLICLE-STIMULATING-HORMONE RESPONSES

FILIPPINI, Antonio;RICCIOLI, ANNA;STEFANINI, Mario;ZIPARO, Elio
1994

Abstract

To study the role of extracellular nucleotides in the regulation of Sertoli cells, the effects of ATP and its analogs on the Ca2+-phospholipid- and cAMP-dependent pathways were tested. Cultured Sertoli cells from immature animals were incubated with ATP or structurally related compounds, and phosphoinositide (PI) turnover or cAMP accumulation was measured. Among the several nucleotide phosphate analogs tested, adenosine 5'-O-(3-thiotriphosphate) was the agonist most potent in stimulating inositol phosphate accumulation. The effects of purine nucleotides on PI turnover were time and concentration dependent. Because nonhydrolizable ATP analogs also stimulated PI turnover, ATP metabolites or metabolic products are not responsible for the observed stimulation. The order of potency of the different ATP analogs [adenosine 5'-O-(3-thiotriphosphate) > ATP congruent to UTP > beta,gamma-methyleneadenosine 5'-triphosphate, 2-methylthio-ATP > adenosine] was consistent with the presence of P-2U receptors (nucleotide receptors) on the surface of the Sertoli cell. Augmented PI turnover was accompanied by a transient increase in Ca2+ concentration, measured in single Sertoli cells loaded with the intracellular Ca2+ indicator fura-2. When used alone, ATP and its analogs did not have a direct effect on cAMP levels in the Sertoli cell. However, ATP or its analogs inhibited FSH-dependent cAMP accumulation by more than 70%. Purine nucleotides also efficiently blocked the effects of FSH distal to cAMP accumulation, because extracellular ATP completely reversed the changes in Sertoli cell shape induced by FSH. The nucleotide-dependent inhibition of cAMP accumulation was blocked by pertussis toxin to a different degree depending on the purine or pirimidine nucleotide used. This indicated that more than one mechanism contributes to the purine nucleotide-dependent inhibition of cAMP accumulation. These data provide evidence that purine nucleotide receptors coupled to multiple pathways are present on the Sertoli cell in culture, and that extracellular ATP has profound biological effects on the FSH responsiveness of the Sertoli cell.
1994
01 Pubblicazione su rivista::01a Articolo in rivista
ACTIVATION OF INOSITOL PHOSPHOLIPID TURNOVER AND CALCIUM SIGNALING IN RAT SERTOLI CELLS BY P2-PURINERGIC RECEPTORS - MODULATION OF FOLLICLE-STIMULATING-HORMONE RESPONSES / Filippini, Antonio; Riccioli, Anna; P., De Cesaris; R., Paniccia; A., Teti; Stefanini, Mario; M., Conti; Ziparo, Elio. - In: ENDOCRINOLOGY. - ISSN 0013-7227. - 134:3(1994), pp. 1537-1545. [10.1210/en.134.3.1537]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/257057
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