Selective activation of Ca2+ influx by extracellular ATP in a pancreatic β-cell line (HIT)

Jean François Geschwind, Marcia Hiriart, Major C. Glennon, Habiba Najafi, Barbara E. Corkey, Franz M. Matschinsky, Marc Prentki

Research output: Contribution to journalArticle

Abstract

The action of exogenous ATP on cytoplasmic free Ca2+ ([Ca2+]i) was studied in insulin secreting cells using fura-2. Stimulation of clonal pancreatic β-cells (HIT) with ATP (range 2-20 μM) evoked a sustained elevation in [Ca2+]i. ATP selectively promoted Ca2+ influx and not Ca2+ mobilization since (1) the effect required external Ca2+ and (2) was observed in cells in which internal stores were depleted with ionomycin (3) the rate of Mn2+ influx, measured as the quenching of the fura-2 signal, was accelerated by ATP. The action of ATP was unaffected by the voltage-sensitive Ca2+ channel blockers nifedipine and verapamil as well as by a depolarizing concentration of K+. The effect on [Ca2+]i was highly specific for ATP since AMP, ADP, adenosine 5′-[γ-thio]triphosphate, adenosine 5′-[β-γ-methylene]triphosphate, GTP and adenosine were ineffective. In normal pancreatic islet cells, both exogenous ATP (range 0.2-2 μM) and ADP induced a transient Ca2+ elevation that did not require external Ca2+. The nucleotide specificity of the effect on [Ca2+]i suggests that ATP activates P2y purinergic receptors in normal β-cells. Thus, ATP evokes a Ca2+ signal in clonal HIT cells and normal islet cells by different transducing systems involving distinct purinoreceptors. A novel mechanism for increasing [Ca2+]i by extracellular ATP is reported in HIT cells, since the nucleotide specificity and the selective activation of Ca2+ influx without mobilization of internal Ca2+ stores cannot be explained by mechanism already described in other cell systems.

Original languageEnglish (US)
Pages (from-to)107-115
Number of pages9
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1012
Issue number1
DOIs
StatePublished - Jun 15 1989
Externally publishedYes

Fingerprint

Adenosine Triphosphate
Cell Line
Islets of Langerhans
Adenosine Diphosphate
Nucleotides
Purinergic Receptors
Ionomycin
Fura-2
Insulin-Secreting Cells
Adenosine Monophosphate
Nifedipine
Verapamil
Guanosine Triphosphate

Keywords

  • (Rat pancreatic β-cell line (HIT))
  • ATP, extracellular
  • Calcium ion
  • cytoplasmic
  • Receptor operated channels

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology
  • Molecular Biology
  • Medicine(all)

Cite this

Geschwind, J. F., Hiriart, M., Glennon, M. C., Najafi, H., Corkey, B. E., Matschinsky, F. M., & Prentki, M. (1989). Selective activation of Ca2+ influx by extracellular ATP in a pancreatic β-cell line (HIT). Biochimica et Biophysica Acta - Molecular Cell Research, 1012(1), 107-115. https://doi.org/10.1016/0167-4889(89)90018-9

Selective activation of Ca2+ influx by extracellular ATP in a pancreatic β-cell line (HIT). / Geschwind, Jean François; Hiriart, Marcia; Glennon, Major C.; Najafi, Habiba; Corkey, Barbara E.; Matschinsky, Franz M.; Prentki, Marc.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1012, No. 1, 15.06.1989, p. 107-115.

Research output: Contribution to journalArticle

Geschwind, JF, Hiriart, M, Glennon, MC, Najafi, H, Corkey, BE, Matschinsky, FM & Prentki, M 1989, 'Selective activation of Ca2+ influx by extracellular ATP in a pancreatic β-cell line (HIT)', Biochimica et Biophysica Acta - Molecular Cell Research, vol. 1012, no. 1, pp. 107-115. https://doi.org/10.1016/0167-4889(89)90018-9
Geschwind, Jean François ; Hiriart, Marcia ; Glennon, Major C. ; Najafi, Habiba ; Corkey, Barbara E. ; Matschinsky, Franz M. ; Prentki, Marc. / Selective activation of Ca2+ influx by extracellular ATP in a pancreatic β-cell line (HIT). In: Biochimica et Biophysica Acta - Molecular Cell Research. 1989 ; Vol. 1012, No. 1. pp. 107-115.
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AB - The action of exogenous ATP on cytoplasmic free Ca2+ ([Ca2+]i) was studied in insulin secreting cells using fura-2. Stimulation of clonal pancreatic β-cells (HIT) with ATP (range 2-20 μM) evoked a sustained elevation in [Ca2+]i. ATP selectively promoted Ca2+ influx and not Ca2+ mobilization since (1) the effect required external Ca2+ and (2) was observed in cells in which internal stores were depleted with ionomycin (3) the rate of Mn2+ influx, measured as the quenching of the fura-2 signal, was accelerated by ATP. The action of ATP was unaffected by the voltage-sensitive Ca2+ channel blockers nifedipine and verapamil as well as by a depolarizing concentration of K+. The effect on [Ca2+]i was highly specific for ATP since AMP, ADP, adenosine 5′-[γ-thio]triphosphate, adenosine 5′-[β-γ-methylene]triphosphate, GTP and adenosine were ineffective. In normal pancreatic islet cells, both exogenous ATP (range 0.2-2 μM) and ADP induced a transient Ca2+ elevation that did not require external Ca2+. The nucleotide specificity of the effect on [Ca2+]i suggests that ATP activates P2y purinergic receptors in normal β-cells. Thus, ATP evokes a Ca2+ signal in clonal HIT cells and normal islet cells by different transducing systems involving distinct purinoreceptors. A novel mechanism for increasing [Ca2+]i by extracellular ATP is reported in HIT cells, since the nucleotide specificity and the selective activation of Ca2+ influx without mobilization of internal Ca2+ stores cannot be explained by mechanism already described in other cell systems.

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