Control of cytosolic free calcium in cultured human pancreatic β-cells occurs by external calcium-dependent and independent mechanisms

Eduardo Rojas, Patricia B. Carroll, Camillo Ricordi, Antonio C. Boschero, Stanko S. Stojilkovic, Illani Atwater

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60 Scopus citations

Abstract

Changes in cytosolic intracellular free Ca2+ ([Ca2+](i)) in response to glucose, glyburide, cholinergic agonists, and elevated [K+](o) (external potassium concentration) were measured in cultured human islet β-cells. In the absence of glucose, the mean resting [Ca2+](i) in single β-cells was 84.5 ± 4.7 nM (n = 86) and remained unchanged in low external [Ca2+](o) (Ca2+ concentration) (<0.2 μM) at 23-25 C. Glucose (5.6-33 mM) induced a slow dose-related [Ca2+](i) rise up to 300.0 ± 50.6 nM (n = 19). This [Ca2+](i) rise always occurred with a delay that varied from cell to cell (~10-120 sec), and the steady state [Ca2+](i) exhibited a sigmoidal dependence on glucose concentration (midpoint at 14.9 mM). The glucose- induced rise in [Ca2+](i) was attenuated by about 62% in low external [Ca2+](o) and was not affected by dantrolene, a drug that inhibits Ca2+ release from the endoplasmic reticulum. In the absence or presence of glucose, cholinergic receptor agonists evoked a biphasic increase in [Ca2+](i) up to 350 nM; the delayed component of the [Ca2+](i) rise was blocked by dantrolene. A rapid elevation of [K+](o) to 40 mM also elicited a biphasic rise in [Ca2+](i), which peaked at about 250 nM and was inhibited by the Ca2+ channel antagonist nifedipine. Glyburide (4 μM) in the absence of glucose also induced a [Ca2+](o)-dependent rise in [Ca2+](i). Increasing the concentration of glucose from 4 to 16.7 mM evoked a biphasic pattern of insulin secretion from perifused isolated islets at 37 C. Finally, in the presence of 4 mM glucose, a cholinergic muscarinic receptor agonist stimulated insulin secretion. A glucose-stimulated [Ca2+](i) rise was also studied at 24 and 37 C in cultured rat islet cells. Our results suggest that the Ca2+ required for glucose-induced and muscarinic agonist-potentiated insulin release enters the cytosol from both extracellular and intracellular Ca2+ stores.

Original languageEnglish (US)
Pages (from-to)1771-1781
Number of pages11
JournalEndocrinology
Volume134
Issue number4
DOIs
StatePublished - Apr 1994
Externally publishedYes

ASJC Scopus subject areas

  • Endocrinology

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