Adenosine 5′-triphosphate (ATP) receptors induce intracellular calcium changes in mouse leydig cells

Elia Martha Pérez-Armendariz, Angel Nadal, Esther Fuentes, David C. Spray

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Cytoplasmic calcium ([Ca 2+] i) changes evoked by adenosine 5 1-triphosphate (ATP) were recorded in cultured individual Leydig cells within 10-18 h after cell dispersion. [Ca 2+] i was monitored using Fura-2AM loaded cells with a digital ratio imaging system. Five micromolars ATP induced biphasic [Ca 2+] i responses in most cells (94%, n=100), characterized by a fast increase from a basal level (126±5 n MSE, n=60 cells) to a peak (5-7 times above basal levels) within seconds, followed by a slow decrease toward a plateau level (2-3 times above basal) within 5 min. The peak phase of the [Ca 2+] i response increased with ATP concentrations (1-100 μM ATP) in a dose-dependent manner with an IC 50 of 5.9±1.2 μM, and it desensitized in a reversible manner with repeated application of 5 μM ATP at <5-min intervals. The [Ca 2+] i peak response was dependent on Ca 2+ release from an intracellular pool, whereas the plateau phase was dependent on extracellular [Ca 2+]. ATP did not appear to induce formation of nonspecific membrane pores, since stimulation for 10 min with ATP (10-100 μM) in the presence of extracellular Lucifer yellow (LY) (5 mg/mL) did not result in dye loading of the cells. [Ca 2+] i transients were elicited by other adenosine nucleotides with an order of potencies (ATP>Adenosine diphosphate [ADP]>Adenosine> Adenosine monophosphate [AMP]) that was compatible with the expression of P 2 receptors. [Ca 2+] i responses were suppressed by the purinergic P 2 receptor antagonist, suramin. These results provide functional evidence for the expression of purinergic P 2 receptors in Leydig cells.

Original languageEnglish (US)
Pages (from-to)239-247
Number of pages9
JournalEndocrine
Volume4
Issue number3
DOIs
StatePublished - Jun 1996

Fingerprint

Leydig Cells
Adenosine Triphosphate
Calcium
Adenosine
Suramin
Adenosine Monophosphate
Adenosine Diphosphate

Keywords

  • gap junctions
  • purinergic receptors
  • secretion
  • Testis

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Endocrinology

Cite this

Adenosine 5′-triphosphate (ATP) receptors induce intracellular calcium changes in mouse leydig cells. / Pérez-Armendariz, Elia Martha; Nadal, Angel; Fuentes, Esther; Spray, David C.

In: Endocrine, Vol. 4, No. 3, 06.1996, p. 239-247.

Research output: Contribution to journalArticle

Pérez-Armendariz, Elia Martha ; Nadal, Angel ; Fuentes, Esther ; Spray, David C. / Adenosine 5′-triphosphate (ATP) receptors induce intracellular calcium changes in mouse leydig cells. In: Endocrine. 1996 ; Vol. 4, No. 3. pp. 239-247.
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abstract = "Cytoplasmic calcium ([Ca 2+] i) changes evoked by adenosine 5 1-triphosphate (ATP) were recorded in cultured individual Leydig cells within 10-18 h after cell dispersion. [Ca 2+] i was monitored using Fura-2AM loaded cells with a digital ratio imaging system. Five micromolars ATP induced biphasic [Ca 2+] i responses in most cells (94{\%}, n=100), characterized by a fast increase from a basal level (126±5 n MSE, n=60 cells) to a peak (5-7 times above basal levels) within seconds, followed by a slow decrease toward a plateau level (2-3 times above basal) within 5 min. The peak phase of the [Ca 2+] i response increased with ATP concentrations (1-100 μM ATP) in a dose-dependent manner with an IC 50 of 5.9±1.2 μM, and it desensitized in a reversible manner with repeated application of 5 μM ATP at <5-min intervals. The [Ca 2+] i peak response was dependent on Ca 2+ release from an intracellular pool, whereas the plateau phase was dependent on extracellular [Ca 2+]. ATP did not appear to induce formation of nonspecific membrane pores, since stimulation for 10 min with ATP (10-100 μM) in the presence of extracellular Lucifer yellow (LY) (5 mg/mL) did not result in dye loading of the cells. [Ca 2+] i transients were elicited by other adenosine nucleotides with an order of potencies (ATP>Adenosine diphosphate [ADP]>Adenosine> Adenosine monophosphate [AMP]) that was compatible with the expression of P 2 receptors. [Ca 2+] i responses were suppressed by the purinergic P 2 receptor antagonist, suramin. These results provide functional evidence for the expression of purinergic P 2 receptors in Leydig cells.",
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