Stochastic NANOG fluctuations allow mouse embryonic stem cells to explore pluripotency

Autor: Petr Svoboda, Hédia Maamar, Martin Moravec, Elsa Abranches, Ana M. V. Guedes, Arjun Raj, Domingos Henrique
Přispěvatelé: Repositório da Universidade de Lisboa
Rok vydání: 2014
Předmět:
Homeobox protein NANOG
Pluripotency
Pluripotent Stem Cells
Transcription
Genetic
Rex1
Stem cells
Biology
Nanog
Time-Lapse Imaging
Cell Line
03 medical and health sciences
Mice
0302 clinical medicine
Animals
Cell Lineage
RNA
Messenger
Lineage priming
Induced pluripotent stem cell
Molecular Biology
reproductive and urinary physiology
Embryonic Stem Cells
030304 developmental biology
Cell Proliferation
Homeodomain Proteins
0303 health sciences
Principal Component Analysis
Stochastic Processes
Lineage markers
Gene Expression Profiling
Nanog Homeobox Protein
Gene Expression Regulation
Developmental
Flow Cytometry
Stem Cells and Regeneration
Molecular biology
Embryonic stem cell
Cell biology
Clone Cells
Kinetics
embryonic structures
Stem cell
biological phenomena
cell phenomena
and immunity
Leukemia inhibitory factor
Gene expression heterogeneity
030217 neurology & neurosurgery
Developmental Biology
Zdroj: Development
ISSN: 1477-9129
Popis: © Published by The Company of Biologists Ltd. All rights reserved.
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.
This work was supported by Fundação para a Ciência e Tecnologia, Portugal [SFRH/ BPD/78313/2011 to E.A., SFRH/BD/80191/2011 to A.M.V.G. and PTDC/SAUOBD/100664/2008]. M.M. and P.S. were supported by The Academy of Sciences of the Czech Republic [project M200521202]. P.S. is a member of the Centre for RNA Biology funded by the Czech Science Foundation [GACR P305/12/G034].
Databáze: OpenAIRE
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