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Almuedo-Castillo, M.

Publications and source records attributed to Almuedo-Castillo, M..

3 recordsLinked to original sources

Optic cup folding is driven by the geometry and tension of the Retinal Pigmented Epithelium (RPE) cells

While optic cup folding is known to involve specific geometrical changes of RPE cells, the precise gene regulatory mechanisms orchestrating the adoption of their highly rigid geometry, and how these contribute to successful folding, remain poorly understood. To address this gap, we investigated how the increase in mechanical tension and maintenance of an elongated geometry depend on the activation of the Wnt/{beta}-catenin and YAP pathways in RPE cells. We demonstrated that interference with these pathways causes folding failure due to a reduction in RPE cellular tension. We also identified transcriptional programs controlled by these pathways that regulate the mechanical properties of the actin cytoskeleton, cell-to-cell and cell-to-ECM adhesions, and endocytosis. Finally, we hypothesized that the LINC complex, which transmits tension between the cell and nuclear membranes, is responsible for the nuclear entry of {beta}-catenin and YAP in a cellular geometry-dependent manner. We combined quantitative imaging, functional analysis, mechanical perturbation assays, and transcriptomic analysis to generate a comprehensive view of how the coordination of mechanosensitive gene expression and changes in cellular geometries drive eye formation.

developmental biology↗

A Yap-dependent transcriptional program directs cell migration for embryo axis assembly

The condensation of the embryo primary axis is a fundamental landmark in the establishment of the vertebrate body plan. Although the complex morphogenetic movements directing cell convergence towards the midline have been described extensively, little is known on how gastrulating cells interpret mechanical cues. Yap proteins are among the best characterized transcriptional mechanotransducers, yet their role in gastrulation has remained elusive. Here we show that the double knockout of yap and its paralog yap1b in medaka results in an axis assembly failure. Quantitative live imaging reveals that mutant cells display reduced displacement and migratory persistence. By characterizing the Yap-dependent transcriptional program, we identified genes involved in cytoskeletal organization and cell-ECM adhesion, rather than in germ layer specification, as direct Yap targets. Dynamic analysis of Tead sensors and downstream targets reveals Yap is acting in migratory cells, and not as a midline beacon, to direct gastrulating precursors trajectories by promoting cortical actin recruitment and focal adhesions assembly. We propose that Yap is engaged in a mechano-regulatory loop that is essential to maintain the directed cell migration sustaining embryo axis formation.

developmental biology↗

Mutation of Vsx genes in zebrafish highlights the robustness of the retinal specification network.

Genetic studies in human and mice have established a dual role for Vsx genes in retina development: an early function in progenitors specification, and a later requirement for bipolar-cells fate determination. Despite their conserved expression patterns, it is currently unclear to which extent Vsx functions are also conserved across vertebrates, as mutant models are available only in mammals. To gain insight into vsx function in teleosts, we have generated vsx1 and vsx2 CRISPR-Cas9 double knockouts (vsxKO) in zebrafish. Our electrophysiological and histological analyses indicate severe visual impairment and bipolar cells depletion in vsxKO larvae, with retinal precursors being rerouted towards photoreceptor or Muller glia fates. Surprisingly, neural retina is properly specified and maintained in mutant embryos, which do not display microphthalmia. We show that although important cis-regulatory remodelling occurs in vsxKO retinas during early specification, this has little impact at a transcriptomic level. Our observations point to genetic redundancy as an important mechanism sustaining the integrity of the retinal specification network, and to Vsx genes regulatory weight varying substantially among vertebrate species. Brief Summary Statement for use in emailed and online tables of content alertsThe mutation of vsx genes in zebrafish confirms a conserved role in bipolar cells specification across vertebrates, but do not interfere with the specification of the neural retina domain. Our data reveal the unexpected robustness of the genetic network sustaining the identity of the neural retina.

developmental biology↗