Top

Published in:

Open Access 01-03-2013 | Article

Autonomous and self-sustained circadian oscillators displayed in human islet cells

Authors: P. Pulimeno, T. Mannic, D. Sage, L. Giovannoni, P. Salmon, S. Lemeille, M. Giry-Laterriere, M. Unser, D. Bosco, C. Bauer, J. Morf, P. Halban, J. Philippe, C. Dibner

Published in: Diabetologia | Issue 3/2013

Abstract

Aims/hypothesis

Following on from the emerging importance of the pancreas circadian clock on islet function and the development of type 2 diabetes in rodent models, we aimed to examine circadian gene expression in human islets. The oscillator properties were assessed in intact islets as well as in beta cells.

Methods

We established a system for long-term bioluminescence recording in cultured human islets, employing lentivector gene delivery of the core clock gene Bmal1 (also known as Arntl)-luciferase reporter. Beta cells were stably labelled using a rat insulin2 promoter fluorescent construct. Single-islet/cell oscillation profiles were measured by combined bioluminescence–fluorescence time-lapse microscopy.

Results

Human islets synchronised in vitro exhibited self-sustained circadian oscillations of Bmal1-luciferase expression at both the population and single-islet levels, with period lengths of 23.6 and 23.9 h, respectively. Endogenous BMAL1 and CRY1 transcript expression was circadian in synchronised islets over 48 h, and antiphasic to REV-ERBα (also known as NR1D1), PER1, PER2, PER3 and DBP transcript circadian profiles. HNF1A and PDX1 exhibited weak circadian oscillations, in phase with the REV-ERBα transcript. Dispersed islet cells were strongly oscillating as well, at population and single-cell levels. Importantly, beta and non-beta cells revealed oscillatory profiles that were well synchronised with each other.

Conclusions/interpretation

We provide for the first time compelling evidence for high-amplitude cell-autonomous circadian oscillators displayed in human pancreatic islets and in dispersed human islet cells. Moreover, these clocks are synchronised between beta and non-beta cells in primary human islet cell cultures.

Available only for authorised users

Reppert SM (2006) A colorful model of the circadian clock. Cell 124:233–236PubMedCrossRef

Dibner C, Schibler U, Albrecht U (2010) The mammalian circadian timing system: organization and coordination of central and peripheral clocks. Annu Rev Physiol 72:517–549PubMedCrossRef

Nagoshi E, Saini C, Bauer C, Laroche T, Naef F, Schibler U (2004) Circadian gene expression in individual fibroblasts: cell-autonomous and self-sustained oscillators pass time to daughter cells. Cell 119:693–705PubMedCrossRef

Balsalobre A, Damiola F, Schibler U (1998) A serum shock induces circadian gene expression in mammalian tissue culture cells. Cell 93:929–937PubMedCrossRef

Kohsaka A, Laposky AD, Ramsey KM et al (2007) High-fat diet disrupts behavioral and molecular circadian rhythms in mice. Cell Metabol 6:414–421CrossRef

Green CB, Takahashi JS, Bass J (2008) The meter of metabolism. Cell 134:728–742PubMedCrossRef

Lamia KA, Storch KF, Weitz CJ (2008) Physiological significance of a peripheral tissue circadian clock. Proc Natl Acad Sci USA 105:15172–15177PubMedCrossRef

Turek FW, Joshu C, Kohsaka A et al (2005) Obesity and metabolic syndrome in circadian clock mutant mice. Science 308:1043–1045PubMedCrossRef

Cho H, Zhao X, Hatori M et al (2012) Regulation of circadian behaviour and metabolism by REV-ERB-α and REV-ERB-β. Nature 485:123–127PubMedCrossRef

10.

Delezie J, Dumont S, Dardente H et al (2012) The nuclear receptor REV-ERBα is required for the daily balance of carbohydrate and lipid metabolism. FASEB J 26:3321–3335PubMedCrossRef

11.

Le Martelot G, Claudel T, Gatfield D et al (2009) REV-ERBalpha participates in circadian SREBP signaling and bile acid homeostasis. PLoS Biology 7:e1000181PubMedCrossRef

12.

Zhang EE, Liu Y, Dentin R et al (2010) Cryptochrome mediates circadian regulation of cAMP signaling and hepatic gluconeogenesis. Nat Med 16:1152–1156PubMedCrossRef

13.

Marcheva B, Ramsey KM, Buhr ED et al (2010) Disruption of the clock components CLOCK and BMAL1 leads to hypoinsulinaemia and diabetes. Nature 466:627–631PubMedCrossRef

14.

Sadacca LA, Lamia KA, deLemos AS, Blum B, Weitz CJ (2011) An intrinsic circadian clock of the pancreas is required for normal insulin release and glucose homeostasis in mice. Diabetologia 54:120–124PubMedCrossRef

15.

Eberhard D, Lammert E (2009) The pancreatic beta-cell in the islet and organ community. Curr Opin Genet Dev 19:469–475PubMedCrossRef

16.

Englund A, Kovanen L, Saarikoski ST et al (2009) NPAS2 and PER2 are linked to risk factors of the metabolic syndrome. J Circadian Rhythms 7:5PubMedCrossRef

17.

Dupuis J, Langenberg C, Prokopenko I et al (2010) New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk. Nat Genet 42:105–116PubMedCrossRef

18.

Stamenkovic JA, Olsson AH, Nagorny CL et al (2012) Regulation of core clock genes in human islets. Metabolism 61:978–985PubMedCrossRef

19.

Bucher P, Mathe Z, Morel P et al (2005) Assessment of a novel two-component enzyme preparation for human islet isolation and transplantation. Transplantation 79:91–97PubMedCrossRef

20.

Ricordi C, Lacy PE, Finke EH, Olack BJ, Scharp DW (1988) Automated method for isolation of human pancreatic islets. Diabetes 37:413–420PubMedCrossRef

21.

Parnaud G, Bosco D, Berney T et al (2008) Proliferation of sorted human and rat beta cells. Diabetologia 51:91–100PubMedCrossRef

22.

Liu AC, Tran HG, Zhang EE, Priest AA, Welsh DK, Kay SA (2008) Redundant function of REV-ERBalpha and beta and non-essential role for Bmal1 cycling in transcriptional regulation of intracellular circadian rhythms. PLoS Genetics 4:e1000023PubMedCrossRef

23.

Salmon P, Oberholzer J, Occhiodoro T, Morel P, Lou J, Trono D (2000) Reversible immortalization of human primary cells by lentivector-mediated transfer of specific genes. Mol Ther 2:404–414PubMedCrossRef

24.

Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien RY (2004) Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nat Biotechnol 22:1567–1572PubMedCrossRef

25.

Toledo JR, Prieto Y, Oramas N, Sanchez O (2009) Polyethylenimine-based transfection method as a simple and effective way to produce recombinant lentiviral vectors. Appl Biochem Biotechnol 157:538–544PubMedCrossRef

26.

Dibner C, Sage D, Unser M et al (2009) Circadian gene expression is resilient to large fluctuations in overall transcription rates. EMBO J 28:123–134PubMedCrossRef

27.

Saini C, Morf J, Stratmann M, Gos P, Schibler U (2012) Simulated body temperature rhythms reveal the phase-shifting behavior and plasticity of mammalian circadian oscillators. Genes Dev 26:567–580PubMedCrossRef

28.

Sage D, Unser M, Salmon P, Dibner C (2010) A software solution for recording circadian oscillator features in time-lapse live cell microscopy. Cell Div 5:17PubMedCrossRef

29.

Luisier F, Blu T, Unser M (2011) Image denoising in mixed Poisson–Gaussian noise. IEEE Trans Image Process 20:696–708PubMedCrossRef

30.

Brown SA, Fleury-Olela F, Nagoshi E et al (2005) The period length of fibroblast circadian gene expression varies widely among human individuals. PLoS Biology 3:e338PubMedCrossRef

31.

Balsalobre A, Brown SA, Marcacci L et al (2000) Resetting of circadian time in peripheral tissues by glucocorticoid signaling. Science 289:2344–2347PubMedCrossRef

32.

Balsalobre A, Marcacci L, Schibler U (2000) Multiple signaling pathways elicit circadian gene expression in cultured Rat-1 fibroblasts. Curr Biol 10:1291–1294PubMedCrossRef

33.

Yoo SH, Yamazaki S, Lowrey PL et al (2004) PERIOD2::LUCIFERASE real-time reporting of circadian dynamics reveals persistent circadian oscillations in mouse peripheral tissues. Proc Natl Acad Sci U S A 101:5339–5346PubMedCrossRef

34.

Asher G, Gatfield D, Stratmann M et al (2008) SIRT1 regulates circadian clock gene expression through PER2 deacetylation. Cell 134:317–328PubMedCrossRef

35.

Miyamoto Y, Sancar A (1999) Circadian regulation of cryptochrome genes in the mouse. Brain Res Mol Brain Res 71:238–243PubMedCrossRef

36.

Sun ZS, Albrecht U, Zhuchenko O, Bailey J, Eichele G, Lee CC (1997) RIGUI, a putative mammalian ortholog of the Drosophila period gene. Cell 90:1003–1011PubMedCrossRef

37.

Liu AC, Welsh DK, Ko CH et al (2007) Intercellular coupling confers robustness against mutations in the SCN circadian clock network. Cell 129:605–616PubMedCrossRef

38.

Allaman-Pillet N, Roduit R, Oberson A et al (2004) Circadian regulation of islet genes involved in insulin production and secretion. Mol Cell Endocrinol 226:59–66PubMedCrossRef

39.

Geiss GK, Bumgarner RE, Birditt B et al (2008) Direct multiplexed measurement of gene expression with color-coded probe pairs. Nat Biotechnol 26:317–325PubMedCrossRef

40.

Gale JE, Cox HI, Qian J, Block GD, Colwell CS, Matveyenko AV (2011) Disruption of circadian rhythms accelerates development of diabetes through pancreatic beta-cell loss and dysfunction. J Biol Rhythm 26:423–433CrossRef

Title: Autonomous and self-sustained circadian oscillators displayed in human islet cells
Authors: P. Pulimeno
T. Mannic
D. Sage
L. Giovannoni
P. Salmon
S. Lemeille
M. Giry-Laterriere
M. Unser
D. Bosco
C. Bauer
J. Morf
P. Halban
J. Philippe
C. Dibner
Publication date: 01-03-2013
Publisher: Springer-Verlag
Published in: Diabetologia / Issue 3/2013
Print ISSN: 0012-186X
Electronic ISSN: 1432-0428
DOI: https://doi.org/10.1007/s00125-012-2779-7

Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin

Prof. Florian Limbourg

Prof. Anoop Chauhan

Developed by: Springer Medicine

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Aims/hypothesis

Methods

Results

Conclusions/interpretation

Please log in to get access to this content

Other articles of this Issue 3/2013

Towards microRNA-based therapeutics for diabetic nephropathy

Reassessment of the putative role of BLK-p.A71T loss-of-function mutation in MODY and type 2 diabetes

The glucagon-like peptide-1 receptor agonist exenatide restores impaired pro-islet amyloid polypeptide processing in cultured human islets: implications in type 2 diabetes and islet transplantation

Glucagon increases circulating fibroblast growth factor 21 independently of endogenous insulin levels: a novel mechanism of glucagon-stimulated lipolysis?

Long-term type 1 diabetes influences haematopoietic stem cells by reducing vascular repair potential and increasing inflammatory monocyte generation in a murine model

Adenovirus-mediated overexpression of Tcfe3 ameliorates hyperglycaemia in a mouse model of diabetes by upregulating glucokinase in the liver

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

At a glance: The STEP trials