Stochastic NANOG fluctuations allow mouse embryonic stem cells to explore pluripotency
Stochastic NANOG fluctuations allow mouse embryonic stem cells to explore pluripotency
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DOI：
10.1242/dev.108910
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发表时间：
2014-07-01
期刊：
影响因子：
4.6
通讯作者：
Henrique, Domingos
中科院分区：
文献类型：
--
作者：
Abranches, Elsa;Guedes, Ana M. V.;Henrique, Domingos
转录因子NANOG的异质性表达与多能干细胞中各种功能状态的存在有关。这种异质性似乎是由单个细胞中Nanog表达的波动引起的，但是仍然缺乏对这些波动及其对多能状态的影响的彻底表征。在这里，我们使用了一种新的荧光报告，以调查小鼠胚胎干细胞（mESC）的NANOG表达的时间动态，并剖析在不同的NANOG状态的mESC的谱系潜力。我们的研究结果表明，随机NANOG波动在mESC中广泛存在，基本上所有表达细胞都显示出NANOG水平的波动，即使在基态条件下培养时（2i培养基）。我们进一步表明，当mESC在标准条件（血清加白血病抑制因子）或基态条件下培养时，波动具有相似的动力学，这意味着NANOG波动是多能状态所固有的。然后，我们比较了在不同条件下生长的低NANOG和高NANOG mESCs的发育潜力，并证实mESCs更容易在低NANOG状态下进入分化。通过基因表达谱的进一步分析显示，低NANOG细胞具有明显的谱系相关基因表达，根据信号环境具有可变的谱。相比之下，高NANOG细胞在不同环境中显示出更稳定的表达谱，具有最小的谱系标志物表达。总而言之，我们的数据支持一个模型，其中随机NANOG波动为mESC提供了探索多种谱系选择的机会，调节其改变功能状态的概率。
Heterogeneous expression of the transcription factor NANOG has been linked to the existence of various functional states in pluripotent stem cells. This heterogeneity seems to arise from fluctuations of Nanog expression in individual cells, but a thorough characterization of these fluctuations and their impact on the pluripotent state is still lacking. Here, we have used a novel fluorescent reporter to investigate the temporal dynamics of NANOG expression in mouse embryonic stem cells (mESCs), and to dissect the lineage potential of mESCs at different NANOG states. Our results show that stochastic NANOG fluctuations are widespread in mESCs, with essentially all expressing cells showing fluctuations in NANOG levels, even when cultured in ground-state conditions (2i media). We further show that fluctuations have similar kinetics when mESCs are cultured in standard conditions (serum plus leukemia inhibitory factor) or ground-state conditions, implying that NANOG fluctuations are inherent to the pluripotent state. We have then compared the developmental potential of low-NANOG and high-NANOG mESCs, grown in different conditions, and confirm that mESCs are more susceptible to enter differentiation at the low-NANOG state. Further analysis by gene expression profiling reveals that low-NANOG cells have marked expression of lineage-affiliated genes, with variable profiles according to the signalling environment. By contrast, high-NANOG cells show a more stable expression profile in different environments, with minimal expression of lineage markers. Altogether, our data support a model in which stochastic NANOG fluctuations provide opportunities for mESCs to explore multiple lineage options, modulating their probability to change functional state.