CoChat
Sign Up
Cited by 174Pancreatic function and diabetesMitochondrial Function and PathologyCalcium signaling and nucleotide metabolism

The Mitochondrial Ca2+ Uniporter MCU Is Essential for Glucose-Induced ATP Increases in Pancreatic β-Cells

Open access
Tarasov, Andrei I., Semplici, Francesca, Ravier, Magalie A., Bellomo, Elisa A., Pullen, Timothy J., Gilon, Patrick, Sekler, Israel, Rizzuto, Rosario +1 more·2012·PLoS ONE
DOI 10.1371/journal.pone.0039722PMC PMC3400633
Open this paper in CoChat — free accountOpen PDF

Abstract

Glucose induces insulin release from pancreatic β-cells by stimulating ATP synthesis, membrane depolarisation and Ca(2+) influx. As well as activating ATP-consuming processes, cytosolic Ca(2+) increases may also potentiate mitochondrial ATP synthesis. Until recently, the ability to study the role of mitochondrial Ca(2+) transport in glucose-stimulated insulin secretion has been hindered by the absence of suitable approaches either to suppress Ca(2+) uptake into these organelles, or to examine the impact on β-cell excitability. Here, we have combined patch-clamp electrophysiology with simultaneous real-time imaging of compartmentalised changes in Ca(2+) and ATP/ADP ratio in single primary mouse β-cells, using recombinant targeted (Pericam or Perceval, respectively) as well as entrapped intracellular (Fura-Red), probes. Through shRNA-mediated silencing we show that the recently-identified mitochondrial Ca(2+) uniporter, MCU, is required for depolarisation-induced mitochondrial Ca(2+) increases, and for a sustained increase in cytosolic ATP/ADP ratio. By contrast, silencing of the mitochondrial Na(+)-Ca(2+) exchanger NCLX affected the kinetics of glucose-induced changes in, but not steady state values of, cytosolic ATP/ADP. Exposure to gluco-lipotoxic conditions delayed both mitochondrial Ca(2+) uptake and cytosolic ATP/ADP ratio increases without affecting the expression of either gene. Mitochondrial Ca(2+) accumulation, mediated by MCU and modulated by NCLX, is thus required for normal glucose sensing by pancreatic β-cells, and becomes defective in conditions mimicking the diabetic milieu.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1371/journal.pone.0039722

Cites

2 of 2 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 47

  1. [1]Rutter GA 2004 Visualising Insulin Secretion. The Minkowski lecture 2004. Diabetologia 47 1861 1872 15551048
  2. [2]Mari A Tura A Natali A Laville M Laakso M 2010 Impaired beta cell glucose sensitivity rather than inadequate compensation for insulin resistance is the dominant defect in glucose intolerance. Diabetologia 53 749 756 20225397
  3. [3]Tarasov AI Nicolson T Riveline JP Taneja TK Baldwin SA 2008 A rare mutation in ABCC8/SUR1 leading to altered KATP channel activity and {beta}-cell glucose sensing is associated with type 2 diabetes mellitus in adults. Diabetes 57 1595 1604 18346985 PMC6101196
  4. [4]Butler AE Janson J Bonner-Weir S Ritzel R Rizza RA 2003 Beta-cell deficit and increased beta-cell apoptosis in humans with type 2 diabetes. Diabetes 52 102 110 12502499
  5. [5]Maechler P Wollheim CB 2001 Mitochondrial function in normal and diabetic beta-cells. Nature 414 807 812 11742413
  6. [6]Rutter GA 2001 Nutrient-secretion coupling in the pancreatic islet b-cell: Recent advances. Molecular Aspects of Medicine 22 247 284 11890977
  7. [7]Kennedy HJ Pouli AE Ainscow EK Jouaville LS Rizzuto R 1999 Glucose generates sub-plasma membrane ATP microdomains in single islet beta-cells. Potential role for strategically located mitochondria. J Biol Chem 274 10224088
  8. [8]Ashcroft FM Harrison DE Ashcroft SJH 1984 Glucose induces closure of single potassium channels in isolated rat pancratic B-cells. Nature Lond 312 446 448 6095103
  9. [9]Wollheim CB Sharp GW 1981 Regulation of insulin release by calcium. Physiol Rev 61 914 973 6117094
  10. [11]Hajnoczky G Robb-Gaspers LD Seitz MB Thomas AP 1995 Decoding of cytosolic calcium oscillations in the mitochondria. Cell 82: 415–424. 0092-8674(95)90430-1 [pii] 7634331
  11. [12]Rutter GA 1990 Ca2+-binding to citrate cycle enzymes. Int J Biochem 22 1081 1088 2289614
  12. [13]Pralong WF Bartley C Wollheim CB 1990 Single islet beta-cell stimulation by nutrients: relationship between pyridine nucleotides, cytosolic Ca2+ and secretion. EMBO J 9 53 60 2403930 PMC551629
  13. [14]Wiederkehr A Szanda G Akhmedov D Mataki C Heizmann CW 2011 Mitochondrial matrix calcium is an activating signal for hormone secretion. Cell Metab 13 601 611 21531342
  14. [15]Rutter GA Theler JM Murgia M Wollheim CB Pozzan T 1993 Stimulated Ca2+ influx raises mitochondrial free Ca2+ to supramicromolar levels in a pancreatic beta-cell line. Possible role in glucose and agonist-induced insulin secretion. J Biol Chem 268 22385 22390 8226749
  15. [16]Eliasson L Renstrom E Ding WG Proks P Rorsman P 1997 Rapid ATP-dependent priming of secretory granules precedes Ca2+-induced exocytosis in mouse pancreatic B-cells. J Physiol London 503 399 412 9306281 PMC1159871
  16. [17]Ainscow EK Rutter GA 2002 Glucose-stimulated oscillations in free cytosolic ATP concentration imaged in single islet b cells: evidnece for a Ca2+-dependent mechanism. Diabetes 51 S162 S170 11815476
  17. [18]Detimary P Gilon P Henquin JC 1998 Interplay between cytoplasmic Ca2+ and the ATP/ADP ratio: a feedback control mechanism in mouse pancreatic islets. Biochem J 333 269 274 9657965 PMC1219582
  18. [20]Nguyen KT Garcia-Chacon LE Barrett JN Barrett EF David G 2009 The Psi(m) depolarization that accompanies mitochondrial Ca2+ uptake is greater in mutant SOD1 than in wild-type mouse motor terminals. Proc Natl Acad Sci U S A 106 2007 2011 19174508 PMC2644154
  19. [21]De Stefani D Raffaello A Teardo E Szabo I Rizzuto R 2011 A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature 476 336 340 21685888 PMC4141877
  20. [22]Baughman JM Perocchi F Girgis HS Plovanich M Belcher-Timme CA 2011 Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature 476 341 345 21685886 PMC3486726
  21. [23]Perocchi F Gohil VM Girgis HS Bao XR McCombs JE 2010 MICU1 encodes a mitochondrial EF hand protein required for Ca(2+) uptake. Nature 467 291 296 20693986 PMC2977980
  22. [24]Palty R Silverman WF Hershfinkel M Caporale T Sensi SL 2010 NCLX is an essential component of mitochondrial Na+/Ca2+ exchange. Proc Natl Acad Sci U S A 107 436 441 20018762 PMC2806722
  23. [25]Filippin L Abad MC Gastaldello S Magalhaes PJ Sandona D 2005 Improved strategies for the delivery of GFP-based Ca2+ sensors into the mitochondrial matrix. Cell Calcium 37 129 136 15589993
  24. [26]Horn R Korn SJ 1992 Prevention of rundown in electrophysiological recording. Methods Enzymol 207 149 55 1382181
  25. [27]Eliasson L Renstrom E Ammala C Berggren PO Bertorello AM 1996 PKC-dependent stimulation of exocytosis by sulfonylureas in pancreatic beta cells. Science 271 813 815 8628999
  26. [28]Berg J Hung YP Yellen G 2009 A genetically encoded fluorescent reporter of ATP: ADP ratio. Nat Methods 6 161 166 19122669 PMC2633436
  27. [29]Tarasov AI Girard CA Ashcroft FM 2006 ATP sensitivity of the ATP-sensitive K+ channel in intact and permeabilized pancreatic beta-cells. Diabetes 55 2446 2454 16936192
  28. [30]Park MK Ashby MC Erdemli G Petersen OH Tepikin AV 2001 Perinuclear, perigranular and sub-plasmalemmal mitochondria have distinct functions in the regulation of cellular calcium transport. EMBO J 20: 1863–1874. 10.1093/emboj/20.8.1863 [doi] PMC125431 11296220 · DOI
  29. [31]Lee B Miles PD Vargas L Luan P Glasco S 2003 Inhibition of mitochondrial Na+-Ca2+ exchanger increases mitochondrial metabolism and potentiates glucose-stimulated insulin secretion in rat pancreatic islets. Diabetes 52 965 973 12663468
  30. [32]Luciani DS Misler S Polonsky KS 2006 Ca2+ controls slow NAD(P)H oscillations in glucose-stimulated mouse pancreatic islets. J Physiol 572 379 392 16455690 PMC1779687
  31. [33]Molina AJ Wikstrom JD Stiles L Las G Mohamed H 2009 Mitochondrial networking protects beta-cells from nutrient-induced apoptosis. Diabetes 58 2303 2315 19581419 PMC2750232
  32. [34]Kohnke R Mei J Park M York DA Erlanson-Albertsson C 2007 Fatty acids and glucose in high concentration down-regulates ATP synthase beta-subunit protein expression in INS-1 cells. Nutr Neurosci 10 273 278 18284036
  33. [35]Gremlich S Bonny C Waeber G Thorens B 1997 Fatty acids decrease IDX-1 expression in rat pancreatic islets and reduce GLUT2, glucokinase, insulin, and somatostatin levels. J Biol Chem 272 30261 30269 9374511
  34. [36]Gilon P Henquin JC 1992 Influence of membrane potential changes on cytoplasmic Ca2+ concentration in an electrically excitable cell, the insulin-secreting pancreatic B-cell. J Biol Chem 267 20713 20720 1400388
  35. [37]Chow RH Lund PE Loser S Panten U Gylfe E 1995 Coincidence of early glucose-induced depolarization with lowering of cytoplasmic Ca2+ in mouse pancreatic beta-cells. J Physiol 485 (Pt 3) 607 617 PMC1158031 7562604
  36. [38]Gilon P Arredouani A Gailly P Gromada J Henquin JC 1999 Uptake and release of Ca2+ by the endoplasmic reticulum contribute to the oscillations of the cytosolic Ca2+ concentration triggered by Ca2+ influx in the electrically excitable pancreatic B-cell. J Biol Chem 274 20197 20205 10400636
  37. [39]Jouaville LS Pinton P Bastianutto C Rutter GA Rizzuto R 1999 Regulation of mitochondrial ATP synthesis by calcium: evidence for a long-term metabolic priming. Proc Natl Acad Sci U S A 96 13807 13812 10570154 PMC24146
  38. [40]Ainscow EK Rutter GA 2001 Mitochondrial priming modifies Ca2+ oscillations and insulin secretion in pancreatic islets. Biochem J 353 175 180 11139378 PMC1221556
  39. [41]Harris DA Das AM 1991 Control of mitochondrial ATP synthesis in the heart. Biochem J 280 561 573 1837214 PMC1130493
  40. [42]Takahashi N Kadowaki T Yazaki Y Ellis-Davies GC Miyashita Y 1999 Post-priming actions of ATP on Ca2+-dependent exocytosis in pancreatic beta cells. Proc Natl Acad Sci U S A 96 760 765 9892707 PMC15210
  41. [43]Tsuboi T da Silva Xavier G, Leclerc I Rutter GA 2003 5′-AMP-activated protein kinase controls insulin-containing secretory vesicle dynamics. J Biol Chem 278 52042 52051 14532293
  42. [44]Parton LE McMillen PJ Shen Y Docherty E Sharpe E 2006 Limited role for SREBP-1c in defective glucose-induced insulin secretion from Zucker diabetic fatty rat islets: a functional and gene profiling analysis. Am J Physiol Endocrinol Metab 291 E982 E994 16772326
  43. [45]Rutter GA Tsuboi T Ravier MA 2006 Ca2+ microdomains and the control of insulin secretion. Cell Calcium 40 539 551 17030367
  44. [46]Ravier MA Rutter GA 2005 Glucose or insulin, but not zinc ions, inhibit glucagon secretion from mouse pancreatic alpha-cells. Diabetes 54 1789 1797 15919801
  45. [47]Luo J Deng ZL Luo X Tang N Song WX 2007 A protocol for rapid generation of recombinant adenoviruses using the AdEasy system. Nat Protoc 2 1236 1247 17546019
  46. [48]Fink C Morgan F Loew LM 1998 Intracellular fluorescent probe concentrations by confocal microscopy. Biophys J 75 1648 1658 9746507 PMC1299837
  47. [49]Tarasov AI Welters HJ Senkel S Ryffel GU Hattersley AT 2006 A Kir6.2 mutation causing neonatal diabetes impairs electrical activity and insulin secretion from INS-1 beta-cells. Diabetes 55 3075 3082 17065345

Cited by

Sorted by citations

Authors
Tarasov, A. I., Semplici, F., Ravier, M. A., Bellomo, E. A., Pullen, T. J., Gilon, P., Sekler, I., Rizzuto, R., & Rutter, G. A.
Affiliations
  • Imperial College London (GB)
  • Centre National de la Recherche Scientifique (FR)
  • UCLouvain (BE)
  • Ben-Gurion University of the Negev (IL)
  • University of Padua (IT)
Corresponding author
Rutter, Guy A.
ORCID
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2012-07-19
Volume
7
Issue
7
Pages
e39722
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0039722
PMC
PMC3400633
OpenAlex
W2072240385
Cited by
174
References
2
Citations by year
2026: 4 · 2025: 12 · 2024: 10 · 2023: 10 · 2022: 8 · 2021: 17 · 2020: 20 · 2019: 8 · 2018: 13 · 2017: 20 · 2016: 9 · 2015: 7 · 2014: 23 · 2013: 10 · 2012: 3
FWCI
13.38
Type
journal article
Language
en
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Access to Libraries (Université catholique de Louvain (UCL), l'Université de Namur (UNamur) and the Université Saint-Louis (USL-B)) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • Figshare (submittedVersion, cc-by)
  • Spiral (Imperial College London) (submittedVersion, cc-by)
  • Padua Research Archive (University of Padova) (submittedVersion, cc-by)
Funders
  • Yale University
  • Wellcome Trust (098424, WT098424AIA, 081958, 098424AIA, 081958/Z/07/Z)
  • Institut National de la Santé et de la Recherche Médicale
  • Medical Research Council (MR/K001981/1)
  • Biotechnology and Biological Sciences Research Council (BB/J015873/1)
Topics
  • Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Physiology › Calcium signaling and nucleotide metabolism
Keywords
mitochondrial calcium uniporter, MCU, mitochondrial calcium uptake, pancreatic beta cells, glucose-stimulated insulin secretion, ATP/ADP ratio, mitochondrial Na+/Ca2+ exchanger, NCLX, glucose detection
MeSH terms
Adenosine Triphosphate, Animals, Calcium, Cells, Cultured, Female, Glucose, Calcium Channels, Gene Silencing, Insulin-Secreting Cells, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review