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research article

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

Pulimeno, P.
•
Mannic, T.
•
Sage, D.  
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2013
Diabetologia

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. 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. 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-ERB alpha (also known as NR1D1), PER1, PER2, PER3 and DBP transcript circadian profiles. HNF1A and PDX1 exhibited weak circadian oscillations, in phase with the REV-ERB alpha 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. 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.

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Type
research article
DOI
10.1007/s00125-012-2779-7
Web of Science ID

WOS:000314531700009

Author(s)
Pulimeno, P.
Mannic, T.
Sage, D.  
Giovannoni, L.
Salmon, P.
Lemeille, S.
Giry-Laterriere, M.
Unser, M.  
Bosco, D.
Bauer, C.
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Date Issued

2013

Publisher

Springer

Published in
Diabetologia
Volume

56

Issue

3

Start page

497

End page

507

Subjects

Beta cells

•

Circadian clocks

•

Human pancreatic islets

•

Time-lapse microscopy

URL

URL

http://bigwww.epfl.ch/publications/pulimeno1301.html

URL

http://bigwww.epfl.ch/publications/pulimeno1301.pdf

URL

http://bigwww.epfl.ch/publications/pulimeno1301.ps
Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
LIB  
Available on Infoscience
March 28, 2013
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/90722
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