Persistent cell-autonomous circadian oscillations in fibroblasts revealed by six-week single-cell imaging of PER2::LUC bioluminescence.
Persistent cell-autonomous circadian oscillations in fibroblasts revealed by six-week single-cell imaging of PER2::LUC bioluminescence.
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DOI：
10.1371/journal.pone.0033334
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Welsh DK
中科院分区：
文献类型：
--
作者：
Leise TL;Wang CW;Gitis PJ;Welsh DK
由于分子反应的随机性质，生物振荡器自然地表现出周期和振幅的随机波动。准确测量噪声振荡器的精度以及整个细胞群体中节律性的周期和强度的异质性，需要足够长的单细胞记录，以充分代表振荡的可变性。我们发现持续的，独立的昼夜节律振荡的时钟基因表达的80个原代成纤维细胞从PER2：：LUC小鼠分离，并保持在体外6个月的6周的生物发光记录。由于节律性的随机性，出现节律的细胞的比例随着检查的间隔长度而增加，当使用3周的窗口时，发现100%的细胞是有节律的。平均周期和振幅在整个6周的记录中非常稳定，精度随着时间的推移而提高。对于单个细胞，周期和振幅的精度与细胞大小和节律振幅相关，但与周期无关，周期的周期间变异性（CV 7.3%）远低于振幅（CV 37%）。时间序列足够长，可以区分每个细胞内的随机波动与细胞间的差异，我们得出结论，细胞确实在周期和节律强度方面表现出显着的异质性，我们使用一种新的统计指标进行测量。此外，随机建模表明，这些单细胞时钟的操作附近的霍普夫分岔，这样的固有噪声增强的振荡，最大限度地减少周期的变化和维持振幅。
Biological oscillators naturally exhibit stochastic fluctuations in period and amplitude due to the random nature of molecular reactions. Accurately measuring the precision of noisy oscillators and the heterogeneity in period and strength of rhythmicity across a population of cells requires single-cell recordings of sufficient length to fully represent the variability of oscillations. We found persistent, independent circadian oscillations of clock gene expression in 6-week-long bioluminescence recordings of 80 primary fibroblast cells dissociated from PER2::LUC mice and kept in vitro for 6 months. Due to the stochastic nature of rhythmicity, the proportion of cells appearing rhythmic increases with the length of interval examined, with 100% of cells found to be rhythmic when using 3-week windows. Mean period and amplitude are remarkably stable throughout the 6-week recordings, with precision improving over time. For individual cells, precision of period and amplitude are correlated with cell size and rhythm amplitude, but not with period, and period exhibits much less cycle-to-cycle variability (CV 7.3%) than does amplitude (CV 37%). The time series are long enough to distinguish stochastic fluctuations within each cell from differences among cells, and we conclude that the cells do exhibit significant heterogeneity in period and strength of rhythmicity, which we measure using a novel statistical metric. Furthermore, stochastic modeling suggests that these single-cell clocks operate near a Hopf bifurcation, such that intrinsic noise enhances the oscillations by minimizing period variability and sustaining amplitude.
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