Frequency-dependent mitochondrial Ca2+ accumulation regulates ATP synthesis in pancreatic cells

Andrei I. Tarasov, Francesca Semplici, Daliang Li, Rosario Rizzuto, Magalie A. Ravier, Patrick Gilon, Guy A. Rutter

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Pancreatic β cells respond to increases in glucose concentration with enhanced metabolism, the closure of ATP-sensitive K+ channels and electrical spiking. The latter results in oscillatory Ca2+ influx through voltage-gated Ca2+ channels and the activation of insulin release. The relationship between changes in cytosolic and mitochondrial free calcium concentration ([Ca2+]cyt and [Ca 2+]mit, respectively) during these cycles is poorly understood. Importantly, the activation of Ca2+-sensitive intramitochondrial dehydrogenases, occurring alongside the stimulation of ATP consumption required for Ca2+ pumping and other processes, may exert complex effects on cytosolic ATP/ADP ratios and hence insulin secretion. To explore the relationship between these parameters in single primary β cells, we have deployed cytosolic (Fura red, Indo1) or green fluorescent protein-based recombinant-targeted (Pericam, 2mt8RP for mitochondria; D4ER for the ER) probes for Ca2+ and cytosolic ATP/ADP (Perceval) alongside patch-clamp electrophysiology. We demonstrate that: (1) blockade of mitochondrial Ca2+ uptake by shRNA-mediated silencing of the uniporter MCU attenuates glucose- and essentially blocks tolbutamide-stimulated, insulin secretion; (2) during electrical stimulation, mitochondria decode cytosolic Ca2+ oscillation frequency as stable increases in [Ca 2+]mit and cytosolic ATP/ADP; (3) mitochondrial Ca 2+ uptake rates remained constant between individual spikes, arguing against activity-dependent regulation ("plasticity") and (4) the relationship between [Ca2+]cyt and [Ca2+] mit is essentially unaffected by changes in endoplasmic reticulum Ca2+ ([Ca2+]ER). Our findings thus highlight new aspects of Ca2+ signalling in β cells of relevance to the actions of both glucose and sulphonylureas.

Original languageEnglish
Pages (from-to)543-554
Number of pages12
JournalPflugers Archiv European Journal of Physiology
Volume465
Issue number4
DOIs
Publication statusPublished - 1 Apr 2013

Keywords

  • ATP
  • Calcium
  • Insulin secretion
  • MCU
  • Mitochondria
  • Oscillation

Fingerprint Dive into the research topics of 'Frequency-dependent mitochondrial Ca<sup>2+</sup> accumulation regulates ATP synthesis in pancreatic cells'. Together they form a unique fingerprint.

  • Cite this