Live imaging of ERK signalling dynamics in differentiating mouse embryonic stem cells

Deathridge, Julia; Antolović, Vlatka; Parsons, Maddy; Chubb, Jonathan R.

doi:10.1242/dev.172940

Cited by 28 publications

(40 citation statements)

References 49 publications

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“…Heterogeneous labelling was observed among the cells (Fig. S3B) (Deathridge et al, 2019;Hamilton and Brickman, 2014), possibly due to heterogeneous levels of NANOG between cells (Chambers et al, 2007) that are controlling Fgf4 secretion, and also owing to possible transient ERK phosphorylation. In the presence of the MEK inhibitor, pERK staining was strongly diminished in ES cells (Fig.…”

Section: Perk Is Present During Postimplantation Development and In Cmentioning

confidence: 99%

Regulation of the ERK signalling pathway in the developing mouse blastocyst

et al. 2019

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Activation of the ERK signalling pathway is essential for the differentiation of the inner cell mass (ICM) during mouse preimplantation development. We show here that ERK phosphorylation occurs in ICM precursor cells, in differentiated primitive endoderm (PrE) cells as well as in the mature, formative state epiblast (Epi). We further show that DUSP4 and ETV5, factors often involved in negative-feedback loops of the FGF pathway, are differently regulated. Whereas DUSP4 presence clearly depends on ERK phosphorylation in PrE cells, ETV5 localises mainly to Epi cells. Unexpectedly, ETV5 accumulation does not depend on direct activation by ERK but requires NANOG activity. Indeed ETV5, like Fgf4 expression, is not present in Nanog mutant embryos. Our results lead us to propose that in pluripotent early Epi cells, NANOG induces the expression of both Fgf4 and Etv5 to enable the differentiation of neighbouring cells into the PrE while protecting the Epi identity from autocrine signalling.

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Section: Perk Is Present During Postimplantation Development and In Cmentioning

confidence: 99%

Regulation of the ERK signalling pathway in the developing mouse blastocyst

et al. 2019

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“…The detection of signal-dependent ERK activity dynamics on short time scales in ESCs was made possible by combining the KTR sensor with high time-resolution recordings. A previous study which examined ERK dynamics upon acute stimulation focused on long term activity and did not resolve the short-timescale oscillations that we report here (Deathridge et al, 2019). These previously undetected dynamics have a modal interpulse interval of approximately 7 minutes (that is, about 8 pulses per hour), and are thus much faster than in any other cell system described so far.…”

Section: Erk Pulses Are More Prevalent Early In the Cell Cyclementioning

confidence: 59%

“…Live-cell ERK activity can be monitored with substrate-based sensors that employ FRET or subcellular localization as read-outs (Komatsu et al, 2011;Regot et al, 2014). Analysis of ERK activity in acutely stimulated ESCs expressing a FRET-based sensor revealed a transient peak of activation that decayed over long timescales (Deathridge et al, 2019). However, the short timescale ERK signaling dynamics in the continuous FGF stimulation regimes required to trigger differentiation of ESCs (Hamilton et al, 2019) remains largely unexplored.…”

Section: Introductionmentioning

confidence: 99%

Intermittent ERK oscillations downstream of FGF in mouse embryonic stem cells

Raina

Morelli

Schröter

2020

Preprint

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Signal transduction networks process extracellular signals to guide cell fate decisions such as to divide, differentiate, or die. These networks can generate characteristic dynamic activities that are shaped by their cell-type specific architecture. The differentiation of pluripotent cells is controlled by FGF/ERK signaling. However, the dynamic activity of the FGF/ERK signaling network in this context remains unexplored. Here we use live cell sensors in wild type and Fgf4 mutant mouse embryonic stem cells to measure ERK dynamic activity in single cells, in response to defined ligand concentrations. We find that ERK activity oscillates in embryonic stem cells. Single cells can transit between oscillatory and non-oscillatory behavior, leading to heterogeneous dynamic activities in the population. Oscillations become more prevalent with increasing FGF4 dose, while maintaining a robust characteristic timescale. Our results suggest that FGF/ERK signaling operates in the vicinity of a transition point between oscillatory and non-oscillatory dynamics in embryonic stem cells.

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“…The Ras/Raf/MEK/ERK signaling pathway is involved in erythropoiesis which mainly promotes growth, differentiation and prevents apoptosis of hematopoietic cells [61][62][63]. MASL1 could induce erythroid differentiation in CD34 cells through the Raf/MEK/ERK signaling pathway [64].…”

Section: Discussionmentioning

confidence: 99%

CRKL but not CRKII Inhibits Erythropoiesis and Megakaryopoiesis of CML via Inactivating Raf/MEK/ERK/Elk-1 Pathway

Guo

Zhang

Yan

et al. 2020

Preprint

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Background: As members of the CT10 regulation of kinase (CRK) adaptor protein family, CRK-like (CRKL) and CRKII are involved in cell proliferation, survival, adhesion, migration and differentiation. However, the exact role and underlying mechanism of CRKL and CRKII in leukemic cell differentiation are still unknown. Methods: Quantitative real-time qPCR (qRT-PCR) was used to detect the expression levels of CRKL and CRKII in chronic myeloid leukemia (CML) patients and complete remission (CR) patients; Western blotting (WB) was used to measure the expression levels of CRKL and CRKII during hemin-induced erythroid differentiation of K562 cells; Benzidine staining, isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis, cDNA microarray assay, qRT-PCR and WB were used to examine the effects of CRKL and CRKII deregulation on erythroid and megakaryocyte differentiation of K562 cells; PD98059 was used to investigate the underlying mechanism of CRKL in erythropoiesis and megakaryopoiesis. Results: CRKL was found to be overexpressed in chronic myeloid leukemia (CML) patients compared with normal samples, while its expression level was lower in CR patients than in corresponding CML patients. The CRKL expression level was significantly decreased during the erythroid differentiation of K562 cells following hemin treatment. Moreover, CRKL downregulation promoted erythroid and megakaryocyte differentiation of K562 cells accompanied by increased expression level of the erythroid differentiation markers γ-globin, glycophorin (GPA) and the megakaryocyte differentiation markers CD41, CD61. Furthermore, gene microarray and iTRAQ quantitative proteomic analysis showed that CRKL downregulation increased hemoglobin (HB) molecules HBD, HBA1, HBA2, HBZ, HBE1, HBG1 and globin transcription factor 1 (GATA1), high-mobility group protein (HMGB2) expression levels. Mechanistically, CRKL inhibited erythroid and megakaryocyte differentiation of K562 cell via inactivating Raf/MEK/ERK/Elk-1 pathway. Conversely, CRKII was only slightly overexpressed in CML patients and had no effect on erythroid differentiation of K562 cells. Conclusions: Taken together, our results demonstrate that CRKL but not CRKII contributes to development, progression, erythropoiesis and megakaryopoiesis of CML, providing novel insights into effective diagnosis and therapy for CML patients.

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Live imaging of ERK signalling dynamics in differentiating mouse embryonic stem cells

Cited by 28 publications

References 49 publications

Regulation of the ERK signalling pathway in the developing mouse blastocyst

Regulation of the ERK signalling pathway in the developing mouse blastocyst

Intermittent ERK oscillations downstream of FGF in mouse embryonic stem cells

CRKL but not CRKII Inhibits Erythropoiesis and Megakaryopoiesis of CML via Inactivating Raf/MEK/ERK/Elk-1 Pathway

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