Generation of pulsatile ERK activity in mouse embryonic stem cells is regulated by Raf activity
Yayoi Toyooka1,2, Kazuhiro Aoki3,4,5, Fumiko Matsukawa Usami6,5
1Division of Embryology, National Institute for Basic Biology, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji-Cho, Okazaki, Aichi, 444-8787, Japan. yayoi.toyooka@cira.kyoto-u.ac.jp.
Scientific Reports
|June 10, 2023
Summary
Extracellular signal-regulated kinase (ERK) activity in embryonic stem cells (ESCs) shows pulsatile dynamics. Raf signaling is crucial for regulating these ERK pulses during differentiation.
Area of Science:
- Cell Biology
- Developmental Biology
- Biochemistry
Background:
- Extracellular signal-regulated kinase (ERK) is a key regulator of cell proliferation and differentiation.
- The ERK signaling pathway is essential for primitive endoderm cell differentiation in embryos and embryonic stem cells (ESCs).
Purpose of the Study:
- To monitor ERK activity in real-time within living undifferentiated and differentiating ESCs.
- To investigate the dynamic patterns of ERK signaling during ESC differentiation.
Main Methods:
- Development of EKAREV-NLS-EB5 ESC lines stably expressing the EKAREV-NLS biosensor.
- Utilizing fluorescence resonance energy transfer (FRET) for live imaging of ERK activity.
- Employing pharmacological inhibition of ERK pathway components.
Main Results:
- ERK activity in ESCs exhibits pulsatile dynamics.
- ESCs were categorized into active (high-frequency pulses) and inactive (no detectable pulses) groups.
- Raf was identified as a critical component influencing ERK pulse patterns.
Conclusions:
- ERK signaling dynamics are pulsatile in ESCs.
- Raf plays a significant role in modulating ERK pulse patterns during differentiation.
- The developed biosensor enables real-time monitoring of ERK activity in ESCs.
