Retinoic Acid Induces Embryonic Stem Cells (ESCs) Transition to 2 Cell-Like State Through a Coordinated Expression of Dux and Duxbl1.

Daniela Tagliaferri,Tiziana Angrisano,Geppino Falco,Antonella Caivano,Martina Addeo,Sabino Russi,Luigi Cerulo,Michele Ceccarelli,Irene Cantone,Pellegrino Mazzone,Vitalba Ruggieri,Mario De Felice,Feliciano Visconte,Luigi Del Vecchio,Teresa M R Noviello

doi:10.3389/fcell.2019.00385

Abstract

Embryonic stem cells (ESCs) are derived from inner cell mass (ICM) of the blastocyst. In serum/LIF culture condition, they show variable expression of pluripotency genes that mark cell fluctuation between pluripotency and differentiation metastate. The ESCs subpopulation marked by zygotic genome activation gene (ZGA) signature, including Zscan4, retains a wider differentiation potency than epiblast-derived ESCs. We have recently shown that retinoic acid (RA) significantly enhances Zscan4 cell population. However, it remains unexplored how RA initiates the ESCs to 2-cell like reprogramming. Here we found that RA is decisive for ESCs to 2C-like cell transition, and reconstructed the gene network surrounding Zscan4. We revealed that RA regulates 2C-like population co-activating Dux and Duxbl1. We provided novel evidence that RA dependent ESCs to 2C-like cell transition is regulated by Dux, and antagonized by Duxbl1. Our suggested mechanism could shed light on the role of RA on ESC reprogramming.

Highlights

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of the blastocyst
To assess how retinoids regulate the transition of ESCs to 2C-like cell population, we grew ESCs in culture medium with or without vitamin A and measured Zscan4 expression level and the percentage of Zscan4+ cells
ESCs cultured for 72 h in retinoids-free N2B27 medium showed a decrease of about 80% of Zscan4 expression compared to N2B27 as measured by quantitative PCR (Figure 1A, left)

Summary

Introduction

Embryonic stem cells (ESCs) are derived from the inner cell mass (ICM) of the blastocyst. In vivo epiblast pluripotency is a transitory state that is maintained in vitro through multiple metastable states that fluctuate between self-renewal and differentiation balance, and display a heterogeneous differentiation potential (Ohtsuka et al, 2012). One of these populations marked by Zscan expression retains wider potency capacity, and it is marked by similar expression of 2-cell stage embryo signature, in particular the activation of MERV-L (murine endogenous retrovirus-like) endogenous retrovirus and the expression of Zscan associated gene family among them Prame, Dux-DuxBl1 Mediate Zscan Metastate. Under standard ESCs culture conditions, about 3-5% of the whole ESCs population expresses Zscan (Zscan4+) which has been shown to mark the ESCs to the so-called “2C-like” transition intermediates (Rodriguez-Terrones et al, 2018)

Methods

Results

Conclusion