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Nanog fluctuations in embryonic stem cells highlight the problem of measurement in cell biology

Nanog fluctuations in embryonic stem cells highlight the problem of measurement in cell biology
Nanog fluctuations in embryonic stem cells highlight the problem of measurement in cell biology
A number of important pluripotency regulators, including the transcription factor Nanog, are observed to fluctuate stochastically in individual embryonic stem (ES) cells. By transiently priming cells for commitment to different lineages, these fluctuations are thought to be important to the maintenance of, and exit from, pluripotency. However, since temporal changes in intracellular protein abundances cannot be measured directly in live cells, fluctuations are typically assessed using genetically engineered reporter cell lines that produce a fluorescent signal as a proxy for protein expression. Here, using a combination of mathematical modeling and experiment, we show that there are unforeseen ways in which widely used reporter strategies can systematically disturb the dynamics they are intended to monitor, sometimes giving profoundly misleading results. In the case of Nanog we show how genetic reporters can compromise the behavior of important pluripotency-sustaining positive feedback loops, and induce a bifurcation in the underlying dynamics that gives rise to heterogeneous Nanog expression patterns in reporter cell lines that are not representative of the wild-type. These findings help explain the range of published observations of Nanog variability and highlight the problem of measurement in live cells.
0006-3495
2641–2652
Smith, Rosanna
1fe5586f-92e9-4658-bd55-cd3eaa176b66
Stumpf, Patrick
dfdb286c-b321-46d3-8ba2-85b3b4a7f092
Ridden, Sonya
f5cd375a-6ba3-4c05-967f-eed2cae66748
Sim, Aaron
b3c3cb91-25f6-4659-8068-4583494ab335
Filippi, Sarah
3796a978-320b-4151-8b68-b29c0f7d70a2
Harrington, Heather
264cceab-f06d-4c01-8ce4-754aeb406d4f
Macarthur, Benjamin
2c0476e7-5d3e-4064-81bb-104e8e88bb6b
Smith, Rosanna
1fe5586f-92e9-4658-bd55-cd3eaa176b66
Stumpf, Patrick
dfdb286c-b321-46d3-8ba2-85b3b4a7f092
Ridden, Sonya
f5cd375a-6ba3-4c05-967f-eed2cae66748
Sim, Aaron
b3c3cb91-25f6-4659-8068-4583494ab335
Filippi, Sarah
3796a978-320b-4151-8b68-b29c0f7d70a2
Harrington, Heather
264cceab-f06d-4c01-8ce4-754aeb406d4f
Macarthur, Benjamin
2c0476e7-5d3e-4064-81bb-104e8e88bb6b

Smith, Rosanna, Stumpf, Patrick, Ridden, Sonya, Sim, Aaron, Filippi, Sarah, Harrington, Heather and Macarthur, Benjamin (2017) Nanog fluctuations in embryonic stem cells highlight the problem of measurement in cell biology. Biophysical Journal, 112 (12), 2641–2652. (doi:10.1016/j.bpj.2017.05.005).

Record type: Article

Abstract

A number of important pluripotency regulators, including the transcription factor Nanog, are observed to fluctuate stochastically in individual embryonic stem (ES) cells. By transiently priming cells for commitment to different lineages, these fluctuations are thought to be important to the maintenance of, and exit from, pluripotency. However, since temporal changes in intracellular protein abundances cannot be measured directly in live cells, fluctuations are typically assessed using genetically engineered reporter cell lines that produce a fluorescent signal as a proxy for protein expression. Here, using a combination of mathematical modeling and experiment, we show that there are unforeseen ways in which widely used reporter strategies can systematically disturb the dynamics they are intended to monitor, sometimes giving profoundly misleading results. In the case of Nanog we show how genetic reporters can compromise the behavior of important pluripotency-sustaining positive feedback loops, and induce a bifurcation in the underlying dynamics that gives rise to heterogeneous Nanog expression patterns in reporter cell lines that are not representative of the wild-type. These findings help explain the range of published observations of Nanog variability and highlight the problem of measurement in live cells.

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Accepted/In Press date: 5 May 2017
e-pub ahead of print date: 20 June 2017
Published date: 20 June 2017
Organisations: Human Development & Health

Identifiers

Local EPrints ID: 404462
URI: http://eprints.soton.ac.uk/id/eprint/404462
ISSN: 0006-3495
PURE UUID: 0a2c0ebd-b3d0-4301-8ac1-f6908dc0fd1d
ORCID for Patrick Stumpf: ORCID iD orcid.org/0000-0003-0862-0290

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Date deposited: 10 Jan 2017 14:06
Last modified: 17 Dec 2019 01:34

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