期刊
DIABETOLOGIA
卷 56, 期 3, 页码 497-507出版社
SPRINGER
DOI: 10.1007/s00125-012-2779-7
关键词
Beta cells; Circadian clocks; Human pancreatic islets; Time-lapse microscopy
资金
- Swiss National Science Foundation [31003A_130070/1]
- EFSD/MSD Basic Research Programme
- Ernst & Lucie Schmidheiny Foundation
- Novartis Foundation for Medico-Biological Sciences Research
- Swiss Life Foundation
- Fondation Romande for the Diabetes Research
- Juvenile Diabetes Research Foundation [31-2008-416]
- Swiss National Science Foundation (SNF) [31003A_130070] Funding Source: Swiss National Science Foundation (SNF)
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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