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Diabetes 51:S162-S170, 2002
© 2002 by the American Diabetes Association, Inc.

Section 4: Oscillations in ß-Cells

Glucose-Stimulated Oscillations in Free Cytosolic ATP Concentration Imaged in Single Islet ß-Cells

Evidence for a Ca2+-Dependent Mechanism

Edward K. Ainscow, and Guy A. Rutter

From the Department of Biochemistry, University of Bristol, Bristol, U.K.

Normal glucose-stimulated insulin secretion is pulsatile, but the molecular mechanisms underlying this pulsatility are poorly understood. Oscillations in the intracellular free [ATP]/[ADP] ratio represent one possible mechanism because they would be expected to cause fluctuations in ATP-sensitive K+ channel activity and hence oscillatory Ca2+ influx. After imaging recombinant firefly luciferase, expressed via an adenoviral vector in single human or mouse islet ß-cells, we report here that cytosolic free ATP concentrations oscillate and that these oscillations are affected by glucose. In human ß-cells, oscillations were observed at both 3 and 15 mmol/l glucose, but the oscillations were of a longer wavelength at the higher glucose concentration (167 vs. 66 s). Mouse ß-cells displayed oscillations in both cytosolic free [Ca2+] and [ATP] only at elevated glucose concentrations, both with a period of 120 s. To explore the causal relationship between [Ca2+] and [ATP] oscillations, the regulation of each was further investigated in populations of MIN6 ß-cells. Incubation in Ca2+-free medium lowered cytosolic [Ca2+] but increased [ATP] in MIN6 cells at both 3 and 30 mmol/l glucose. Removal of external Ca2+ increased [ATP], possibly by decreasing ATP consumption by endoplasmic reticulum Ca2+-ATPases. These results allow a model to be constructed of the ß-cell metabolic oscillator that drives nutrient-induced insulin secretion.


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