Cell-Cycle-Dependent ERK Signaling Dynamics Direct Fate Specification in the Mammalian Preimplantation Embryo

Michael J Pokrass; Kathleen A Ryan; Tianchi Xin; Brittany Pielstick; Winston Timp; Valentina Greco; Sergi Regot

doi:10.1016/j.devcel.2020.09.013

Cell-Cycle-Dependent ERK Signaling Dynamics Direct Fate Specification in the Mammalian Preimplantation Embryo

Dev Cell. 2020 Nov 9;55(3):328-340.e5. doi: 10.1016/j.devcel.2020.09.013. Epub 2020 Oct 21.

Authors

Michael J Pokrass¹, Kathleen A Ryan², Tianchi Xin³, Brittany Pielstick⁴, Winston Timp⁵, Valentina Greco³, Sergi Regot⁶

Affiliations

¹ Department Molecular Biology and Genetics, the Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department Oncology, the Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Biochemistry, Cellular, and Molecular Biology Graduate Program, Baltimore, MD, USA.
² Department Molecular Biology and Genetics, the Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department Oncology, the Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
³ Genetics Department, Yale School of Medicine, New Haven, CT 06520, USA.
⁴ Biochemistry, Cellular, and Molecular Biology Graduate Program, Baltimore, MD, USA; Biomedical Engineering Department, the Johns Hopkins University, Baltimore, MD 21218, USA.
⁵ Biomedical Engineering Department, the Johns Hopkins University, Baltimore, MD 21218, USA.
⁶ Department Molecular Biology and Genetics, the Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA; Department Oncology, the Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. Electronic address: sregot@jhmi.edu.

Abstract

Despite the noisy nature of single cells, multicellular organisms robustly generate different cell types from one zygote. This process involves dynamic cross regulation between signaling and gene expression that is difficult to capture with fixed-cell approaches. To study signaling dynamics and fate specification during preimplantation development, we generated a transgenic mouse expressing the ERK kinase translocation reporter and measured ERK activity in single cells of live embryos. Our results show primarily active ERK in both the inner cell mass and trophectoderm cells due to fibroblast growth factor (FGF) signaling. Strikingly, a subset of mitotic events results in a short pulse of ERK inactivity in both daughter cells that correlates with elevated endpoint NANOG levels. Moreover, endogenous tagging of Nanog in embryonic stem cells reveals that ERK inhibition promotes enhanced stabilization of NANOG protein after mitosis. Our data show that cell cycle, signaling, and differentiation are coordinated during preimplantation development.

Keywords: ERK; NANOG; blastocyst; cell cycle; embryonic stem cells; preimplantation development; signaling dynamics.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Animals
Blastocyst / cytology*
Blastocyst / enzymology*
Cell Cycle*
Cell Lineage*
Germ Layers / cytology
Humans
MAP Kinase Signaling System*
Mammals / embryology*
Mice
Mitosis
Models, Biological
Nanog Homeobox Protein / metabolism
Protein Stability
Reproducibility of Results

Substances

Nanog Homeobox Protein

Grants and funding

R35 GM133499/GM/NIGMS NIH HHS/United States