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Prediction and control of symmetry breaking in embryoid bodies by environment and signal integration

Naor Sagy, Shaked Slovin, Maya Allalouf, Maayan Pour, Gaya Savyon, Jonathan Boxman, View ORCID ProfileIftach Nachman
doi: https://doi.org/10.1101/506543
Naor Sagy
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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Shaked Slovin
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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Maya Allalouf
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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Maayan Pour
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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Gaya Savyon
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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Jonathan Boxman
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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Iftach Nachman
Department of Biochemistry and Molecular Biology, Tel Aviv University, Israel
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  • ORCID record for Iftach Nachman
  • For correspondence: iftachn@tauex.tau.ac.il
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Abstract

During early embryogenesis, mechanical signals, localized biochemical signals and neighboring cell layers interaction coordinate around anteroposterior axis determination and symmetry breaking. Deciphering their relative roles, which are hard to tease apart in vivo, will enhance our understanding of how these processes are driven. In recent years, in vitro 3D models of early mammalian development, such as embryoid bodies (EBs) and gastruloids, were successful in mimicking various aspects of the early embryo, providing high throughput accessible systems for studying the basic rules shaping cell fate and morphology during embryogenesis. Using Brachyury (Bry), a primitive streak and mesendoderm marker in EBs, we study how contact, biochemical and neighboring cell cues affect the positioning of a primitive streak-like locus, determining the AP axis. We show that a Bry-competent layer must be formed in the EB before Bry expression initiates, and that Bry onset locus selection depends on contact points of the EB with its surrounding. We can maneuver Bry onset to occur at a specific locus, a few loci, or in an isotropic peripheral pattern. By spatially separating contact and biochemical signal sources, we show these two modalities can be integrated by the EB to generate a single Bry locus. Finally, we show Foxa2+ cells are predictive of the future location of Bry onset, demonstrating an earlier symmetry-breaking event. By delineating the temporal signaling pathway dependencies of Bry and Foxa2, we were able to selectively abolish either, or spatially decouple the two cell types during EB differentiation. These findings demonstrate multiple inputs integration during an early developmental process, and may prove valuable in directing in vitro differentiation.

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Posted December 26, 2018.
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Prediction and control of symmetry breaking in embryoid bodies by environment and signal integration
Naor Sagy, Shaked Slovin, Maya Allalouf, Maayan Pour, Gaya Savyon, Jonathan Boxman, Iftach Nachman
bioRxiv 506543; doi: https://doi.org/10.1101/506543
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Prediction and control of symmetry breaking in embryoid bodies by environment and signal integration
Naor Sagy, Shaked Slovin, Maya Allalouf, Maayan Pour, Gaya Savyon, Jonathan Boxman, Iftach Nachman
bioRxiv 506543; doi: https://doi.org/10.1101/506543

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