Biology subjects

Alegria-Prevot, O.

Publications and source records attributed to Alegria-Prevot, O..

4 recordsLinked to original sources

bioRxiv2025

Actin and myosin dynamics during epithelial remodeling in avian gastrulation

Epithelial remodeling is powered by contractile forces exerted by the actomyosin cytoskeleton. In invertebrates, pulsatile contractile flows of the medio-apical actomyosin cortex have been shown to be critical in promoting junction contraction that ultimately drive cell rearrangements, apical constriction, and cell extrusion. However, how actomyosin dynamics drives epithelial remodeling in amniotes remains poorly understood. In this study, we generated transgenic quail lines reporting actin and myosin and investigate their dynamics in gastrulating embryos. We show that during this process, epithelial remodeling events are closely associated with the contraction of junctional myosin. Although we observe medio-apical contractile flows, those appear to contribute to junction contraction by promoting junctional myosin recruitment. Notably, by characterizing live and apoptotic cell extrusions and their associated actomyosin dynamics, we provide new insights into the cellular processes underlying primitive streak formation and the emergence of germ layers.

developmental biology↗

bioRxiv2025

Live imaging and functional characterization of the avian hypoblast redefine the mechanisms of primitive streak induction

In birds and mammals, the formation of the primitive streak, the hallmark of the primary axis and site of gastrulation, is thought to occur when an anterior displacement of the hypoblast (visceral endoderm in mice) lifts its inhibition on the posterior epiblast, enabling the activation of NODAL signaling. Although the anterior movement of the murine visceral endoderm is well documented, the dynamics of the avian hypoblast remain poorly understood. Here, using live imaging and quantitative image analysis, we find that the hypoblast is mechanically coupled to the epiblast and does not migrate away from its posterior end. Instead, the hypoblast moves and deforms passively, in response to the forces transmitted from the epiblast that shape the primitive streak, after its induction. Furthermore, we show that the posterior hypoblast does not exert an inhibitory effect on the epiblast but instead expresses NODAL, which activates primitive streak formation. NODAL concomitantly regulates gene expression in the hypoblast, patterning it along the anteroposterior axis. Our results thus redefine how the primary axis is established in avians, demonstrating that the displacement of the hypoblast and its concomitant anteroposterior patterning are consequences -- rather than drivers -- of primitive streak induction, downstream of NODAL signaling. Essential summaryNODAL asymmetric expression in the hypoblast activates primitive streak formation in the avian embryo.

developmental biology↗

bioRxiv2024

CRISPR/Cas9-based somatic knock-in of reporters in the avian embryo in ovo

Gene editing and protein tagging are at the heart of modern developmental and cell biology. The advent of CRISPR/Cas9 based methods offers the possibility to develop customized approaches for genomic manipulations in non-classical experimental models. Here, we show that highly efficient somatic knock-ins of long DNA fragments can be achieved in the developing chick neural tube in ovo. We compare different types of repair matrices and different methods for the delivery of the CRISPR/Cas9 machinery, and find that an all plasmid-based approach and short arms of homology provide an easy and efficient method to achieve high frequencies of knock-in insertions with virtually no background signal. We use this method to target fluorescent reporters and dynamically monitor the subcellular distribution of endogenously expressed tagged proteins, as well as to insert the Gal4-VP16 transcription factor or the Cre recombinase at specific loci to label neural sub-populations in the chick embryonic spinal cord. Finally, we show that the method can also be applied to target the epiblast and somitic mesoderm.

developmental biology↗

bioRxiv2021

Self-organized tissue mechanics underlie embryonic regulation

Early amniote development is a highly regulative and self-organized process, capable to adapt to interference through cell-cell interactions, which are widely believed to be mediated by molecules. Analyzing intact and mechanically perturbed avian embryos, we show that the mechanical forces that drive embryogenesis self-organize in an analog of Turings molecular reaction-diffusion model, with contractility locally self-activating and the ensuing tension acting as a long-range inhibitor. This mechanical feedback governs the persistent pattern of tissue flows that shape the embryo and steers the concomitant emergence of embryonic territories by modulating gene expression, ensuring the formation of a single embryo under normal conditions, yet allowing the emergence of multiple, well-proportioned embryos upon perturbations. Thus, mechanical forces are a central signal in embryonic self-organization, feeding back onto gene expression to canalize both patterning and morphogenesis.

developmental biology↗