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  • Title: Circadian rhythmicity in murine pre-adipocyte and adipocyte cells.
    Author: Otway DT, Frost G, Johnston JD.
    Journal: Chronobiol Int; 2009 Oct; 26(7):1340-54. PubMed ID: 19916835.
    Abstract:
    Adipose tissue is central to metabolic homeostasis, signaling in part through the secretion of molecules termed adipokines. Circadian rhythms play an important role in adipose physiology, with plasma adipokine concentration and approximately 20 % of the murine adipose transcriptome undergoing 24 h variation. However, due to the heterogeneity of adipose tissue and rhythmical input from both neuronal and humoral signals, the cellular basis of adipose rhythms is unclear. We tested the hypothesis that adipocyte cells contain a circadian clock that drives rhythmic mRNA expression and adipokine secretion. From the murine pre-adipocyte 3T3-L1 cell line, we generated populations of both pre-adipocytes and differentiated adipocytes. Cells were then treated with a 2 h serum pulse and sampled every 4 h over a 48 h period. Expression of clock gene, 'metabolic' gene (PPARalpha, PPARgamma, SREBP1), and adipokine mRNA was analyzed by quantitative real-time PCR, and secretion of the adipokines leptin and adiponectin was measured in culture medium from differentiated adipocytes. In pre-adipocytes, we observed robust rhythms of clock genes Per2, Rev-erbalpha, and Dbp, but not of Per1, Cry1, Bmal1, or any of the 'metabolic' genes. Adipocytes produced similar temporal profiles of mRNA expression, albeit with a markedly reduced amplitude of Per2 and Dbp rhythms. Despite no circadian rhythm of adipokine mRNA expression, leptin accumulation in the culture medium suggested circadian control of leptin secretion from adipocytes. Adiponectin secretion showed temporal variation, but without any apparent circadian rhythmicity. Our data, therefore, suggest that an endogenous adipocyte clock controls the rhythmic expression of only a subset of genes that are reported to exhibit 24 h rhythmicity in murine adipose tissue. Moreover, secretion of leptin may also be regulated by the adipocyte clock.
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