Mesoderm is required for coordinated cell movements within zebrafish neural plate in vivo.

BACKGROUND: Morphogenesis of the zebrafish neural tube requires the coordinated movement of many cells in both time and space. A good example of this is the movement of the cells in the zebrafish neural plate as they converge towards the dorsal midline before internalizing to form a neural keel. How these cells are regulated to ensure that they move together as a coherent tissue is unknown. Previous work in other systems has suggested that the underlying mesoderm may play a role in this process but this has not been shown directly in vivo. RESULTS: Here we analyze the roles of subjacent mesoderm in the coordination of neural cell movements during convergence of the zebrafish neural plate and neural keel formation. Live imaging demonstrates that the normal highly coordinated movements of neural plate cells are lost in the absence of underlying mesoderm and the movements of internalization and neural tube formation are severely disrupted. Despite this, neuroepithelial polarity develops in the abnormal neural primordium but the resulting tissue architecture is very disorganized. CONCLUSIONS: We show that the movements of cells in the zebrafish neural plate are highly coordinated during the convergence and internalization movements of neurulation. Our results demonstrate that the underlying mesoderm is required for these coordinated cell movements in the zebrafish neural plate in vivo

Zebrafish as an emerging model to study estrogen receptors in neural development

Estrogens induce several regulatory signals in the nervous system that are mainly mediated through estrogen receptors (ERs). ERs are largely expressed in the nervous system, yet the importance of ERs to neural development has only been elucidated over the last decades. Accumulating evidence shows a fundamental role for estrogens in the development of the central and peripheral nervous systems, hence, the contribution of ERs to neural function is now a growing area of research. The conservation of the structure of the ERs and their response to estrogens make the zebrafish an interesting model to dissect the role of estrogens in the nervous system. In this review, we highlight major findings of ER signaling in embryonic zebrafish neural development and compare the similarities and differences to research in rodents. We also discuss how the recent generation of zebrafish ER mutants, coupled with the availability of several transgenic reporter lines, its amenability to pharmacological studies and in vivo live imaging, could help us explore ER function in embryonic neural development

Étude d'un gène hsp40 au cours de la régénération de la nageoire caudale et du développement embryonnaire chez le poisson zèbre (Danio rerio)

PARIS7-Bibliothèque centrale (751132105) / SudocSudocFranceF

La régénération des appendices chez les vertébrés: un modèle expérimental ancien pour étudier les cellules souches chez l’adulte

La recherche sur les cellules souches laisse entrevoir d’extraordinaires possibilités de traitement des maladies dégénératives. En effet, la capacité de pouvoir dériver des cellules totipotentes à partir d’embryons humains donne la possibilité de développer une médecine régénérative, mais pose également le problème du statut de l’embryon qui, dans ce cas, est considéré comme matériel thérapeutique. Une alternative à l’utilisation des cellules souches embryonnaires humaines est l’utilisation de cellules souches prélevées chez l’adulte. Mais, dans un cas comme dans l’autre, nos connaissances sur les cellules totipotentes ou pluripotentes sont insuffisantes et de nombreuses questions doivent être résolues avant que l’on ne maîtrise la sélection et la différenciation de ces cellules dans un type cellulaire donné. Quelles sont les caractéristiques moléculaires d’une cellule souche adulte? Quels sont les mécanismes sous-jacents à la re-programmation d’une cellule? Quels sont les signaux qui contrôlent la multiplication et la différenciation des cellules souches? Un travail de recherche fondamentale est nécessaire pour éclaircir ces différents points. Dans ce contexte, la régénération des appendices chez les vertébrés offre un terrain d’investigation intéressant. Cet article se propose de faire le point sur nos connaissances concernant la régénération des pattes chez les tétrapodes et des nageoires chez les poissons.The application of stem cell therapy to cure degenerative diseases offers immense possibilities, but the research in this field is the subject of ethical debates raised by the question of destructive research on early human embryos. Stem cells taken in the adult constitute an alternative to human embryonic stem cells, but our knowledge on totipotent or pluripotent cells is currently insufficient. Furthermore, many questions must be solved before selection and differentiation of these cells in a given cellular type can be controlled on a routine basis. What are the molecular characteristics of an adult stem cell? What are the mechanisms involved in cell reprogramming? Which signals control stem cell replication and differentiation? Basic research activities must be carried out in order to clarify all these points. In this context, the regeneration of vertebrate appendages provides a model for this type of research. The regeneration process is defined by both the morphological and functional reconstruction of a part of a living organism, which has previously been destroyed. But why are some vertebrates able to regenerate complex structures and others apparently not? Among most vertebrates, the capacity to regenerate is limited to some tissues. It is however possible to observe the regeneration of appendages (limb, tail, fin, jaw, etc.) among several amphibians and fish. This regeneration leads to re-forming of the amputated part with a complete restoration of its shape, segmentation and function. Why is the amputation of limbs not followed by regeneration in mammals and birds: absence of stem cells, absence of recruitment signals for these cells, or absence of signal receptivity? This review constitutes a report on the current understanding of the basis of on regeneration of legs in tetrapods and of fins in fish with an emphasis in the role of the nervous system in this process

Rho GTPases Signaling in Zebrafish Development and Disease

Cells encounter countless external cues and the specificity of their responses is translated through a myriad of tightly regulated intracellular signals. For this, Rho GTPases play a central role and transduce signals that contribute to fundamental cell dynamic and survival events. Here, we review our knowledge on how zebrafish helped us understand the role of some of these proteins in a multitude of in vivo cellular behaviors. Zebrafish studies offer a unique opportunity to explore the role and more specifically the spatial and temporal dynamic of Rho GTPases activities within a complex environment at a level of details unachievable in any other vertebrate organism

Neuronal Ndrg4 Is Essential for Nodes of Ranvier Organization in Zebrafish

International audienceAxon ensheathment by specialized glial cells is an important process for fast propagation of action potentials. The rapid electrical conduction along myelinated axons is mainly due to its saltatory nature characterized by the accumulation of ion channels at the nodes of Ranvier. However, how these ion channels are transported and anchored along axons is not fully understood. We have identified N-myc downstream-regulated gene 4, ndrg4, as a novel factor that regulates sodium channel clustering in zebrafish. Analysis of chimeric larvae indicates that ndrg4 functions autonomously within neurons for sodium channel clustering at the nodes. Molecular analysis of ndrg4 mutants shows that expression of snap25 and nsf are sharply decreased, revealing a role of ndrg4 in controlling vesicle exocytosis. This uncovers a previously unknown function of ndrg4 in regulating vesicle docking and nodes of Ranvier organization, at least through its ability to finely tune the expression of the t-SNARE/NSF machinery

Wnt/beta-catenin signaling is an essential and direct driver of myelin gene expression and myelinogenesis.

International audienceWnt/beta-catenin signaling plays a major role in the development of the nervous system and contributes to neuronal plasticity. However, its role in myelination remains unclear. Here, we identify the Wnt/beta-catenin pathway as an essential driver of myelin gene expression. The selective inhibition of Wnt components by small interfering RNA or dominant-negative forms blocks the expression of myelin protein zero (MPZ) and peripheral myelin protein 22 (PMP22) in mouse Schwann cells and proteolipid protein in mouse oligodendrocytes. Moreover, the activation of Wnt signaling by recombinant Wnt1 ligand increases by threefold the transcription of myelin genes and enhances the binding of beta-catenin to T-cell factor/lymphoid-enhancer factor transcription factors present in the vicinity of the MPZ and PMP22 promoters. Most important, loss-of-function analyses in zebrafish embryos show, in vivo, a key role for Wnt/beta-catenin signaling in the expression of myelin genes and in myelin sheath compaction, both in the peripheral and central nervous systems. Inhibition of Wnt/beta-catenin signaling resulted in hypomyelination, without affecting Schwann cell and oligodendrocyte generation or axonal integrity. The present findings attribute to Wnt/beta-catenin pathway components an essential role in myelin gene expression and myelinogenesis

ndrg4 is not required for axonal outgrowth or early Schwann cell development.

<p>Acetylated tubulin expression in control (A), ndrg4 mutant (C) and morphant (B) embryos at 4 dpf showing the PLLn nerve. Scale bar = 45μm. (d-F) Whole mount <i>in situ</i> hybridization of a (d) control embryo, ndrg4 mutant (F) and ndrg4 morphant (E) showing <i>sox10</i> expression in PLLn SCs (arrows) at 3 dpf. Scale bar = 200μm. Lateral view of a control foxd3::GFP embryo (G), a ndrg4 morphant (H) at 3 dpf showing SCs (arrows) along the PLLn. Transmission electron micrographs showing cross-section through (I) control and ndrg4 mutant (J). Control PLLn shows an average of 10.7 myelinated axons (blue asterisks). (J) An average of 5.36 myelinated axons (blue asterisks) is observed in the ndrg4 mutant’s PLLn. (S: Schwann cell). Scale bars = 0.5μm. (K,L) Quantification of the total number of axons and the number myelinated axons in controls, ndrg4 mutants and ndrg4 morphants. NS: Non Significant.</p

ndrg4 is required for sodium channel and neurofascin clustering in the peripheral nervous system.

<p>(A-C) Lateral view of <i>mbp</i> RNA expression in control (A), ndrg4 mutant (B) and morphant (C) embryos at 4 dpf. Arrows indicate <i>mbp</i>-expressing cells along the PLLn. Scale bars = 200μm. (D-L) Acetylated tubulin (ac tub; red) and sodium channels (NaCh; green) immunohistochemistry of a (D-F) control, ndrg4 mutant (G-I) and ndrg4 morphant (J-L) PLLn at 4 dpf. Scale bars = 5μm. (M) High magnification of three nodes of Ranvier (arrowheads) from a control nerve. Scale bar = 100nm. (N) A significant decrease in the number of sodium channels clusters is observed in ndrg4 mutants and morphants in comparison to controls (p<0.001). Acetylated tubulin (ac tub; red) and FIGQY (green) immunohistochemistry of a (D’-F’) control, ndrg4 mutant (G’-I’) and ndrg4 morphant (J’-L’) PLLn at 4dpf. Scale bars = 5 μm. (O,P) A significant decrease in the number of FIGQY labeled clusters is observed in ndrg4 mutants and morphants in comparison to controls (p<0.001).</p