Ewing Sarcoma Protein Ewsr1 Maintains Mitotic Integrity and Proneural Cell Survival in the Zebrafish Embryo

BACKGROUND:The Ewing sarcoma breakpoint region 1 gene (EWSR1), also known as EWS, is fused to a number of different partner genes as a result of chromosomal translocation in diverse sarcomas. Despite the involvement of EWSR1 in these diverse sarcomas, the in vivo function of wild type EWSR1 remains unclear. PRINCIPAL FINDINGS:We identified two zebrafish EWSR1 orthologues, ewsr1a and ewsr1b, and demonstrate that both genes are expressed maternally, and are expressed ubiquitously throughout zebrafish embryonic development. Morpholino induced knockdown of both zebrafish ewsr1 genes led to mitotic defects with multipolar or otherwise abnormal mitotic spindles starting from the bud stage (10 hour post-fertilization (hpf)). The abnormalities in mitotic spindles were followed by p53-mediated apoptosis in the developing central nervous system (CNS) leading to a reduction in the number of proneural cells, disorganization of neuronal networks, and embryonic lethality by 5 days post-fertilization. siRNA silencing of EWSR1 in Hela cells resulted in mitotic defects accompanied by apoptotic cell death, indicating that the role of EWSR1 is conserved between zebrafish and human. CONCLUSIONS:Ewsr1 maintains mitotic integrity and proneural cell survival in early zebrafish development

In vivo cell biology using Gal4-mediated multicolor subcellular labeling in zebrafish

The behavior of a cell is determined by the interplay of its subcellular components. Thus, being able to simultaneously visualize several organelles inside cells within the natural context of a living organism could greatly enhance our understanding of developmental processes. We have established a Gal4-based system for the simultaneous and cell type specific expression of multiple subcellular labels in transparent zebrafish embryos. This system offers the opportunity to follow intracellular developmental processes in a live vertebrate organism using confocal fluorescence time-lapse microscopy. Using this approach we recently showed that the centrosome neither persistently leads migration nor determines the site of axonogenesis in migrating neurons in the zebrafish cerebellum in vivo. Here we present additional in vivo findings about the centrosomal and microtubule dynamics of neuroepithelial cells during mitotic cleavages at early neural tube stages

Tension-oriented cell divisions limit anisotropic tissue tension in epithelial spreading during zebrafish epiboly

International audienceEpithelial spreading is a common and fundamental aspect of various developmental and disease-related processes such as epithelial closure and wound healing. A key challenge for epithelial tissues undergoing spreading is to increase their surface area without disrupting epithelial integrity. Here we show that orienting cell divisions by tension constitutes an efficient mechanism by which the enveloping cell layer (EVL) releases anisotropic tension while undergoing spreading during zebrafish epiboly. The control of EVL cell-division orientation by tension involves cell elongation and requires myosin II activity to align the mitotic spindle with the main tension axis. We also found that in the absence of tension-oriented cell divisions and in the presence of increased tissue tension, EVL cells undergo ectopic fusions, suggesting that the reduction of tension anisotropy by oriented cell divisions is required to prevent EVL cells from fusing. We conclude that cell-division orientation by tension constitutes a key mechanism for limiting tension anisotropy and thus promoting tissue spreading during EVL epiboly

Mitotic cell rounding and epithelial thinning regulate lumen growth and shape

Many organ functions rely on epithelial cavities with particular shapes. Morphogenetic anomalies in these cavities lead to kidney, brain or inner ear diseases. Despite their relevance, the mechanisms regulating lumen dimensions are poorly understood. Here, we perform live imaging of zebrafish inner ear development and quantitatively analyse the dynamics of lumen growth in 3D. Using genetic, chemical and mechanical interferences, we identify two new morphogenetic mechanisms underlying anisotropic lumen growth. The first mechanism involves thinning of the epithelium as the cells change their shape and lose fluids in concert with expansion of the cavity, suggesting an intra-organ fluid redistribution process. In the second mechanism, revealed by laser microsurgery experiments, mitotic rounding cells apicobasally contract the epithelium and mechanically contribute to expansion of the lumen. Since these mechanisms are axis specific, they not only regulate lumen growth but also the shape of the cavity.This work has been supported by grants BFU2011-27006 from MICINN and XVICN from Fundación Areces to B.A

N-cadherin–mediated cell adhesion restricts cell proliferation in the dorsal neural tube

N-cadherin (N-cad), an adherens junction (AJ) component, restricts neural progenitor division and couples progenitor cell-cycle exit and differentiation. The effect of N-cad on cell proliferation is mediated by ectopic activation of Hedgehog (Hh) signaling. Hh itself promotes AJ assembly, suggesting a reciprocal interaction between AJs and Hh signaling

Stretching Morphogenesis of the Roof Plate and Formation of the Central Canal

10.1371/journal.pone.0056219PLoS ONE82