Control of convergent yolk syncytial layer nuclear movement in zebrafish (original) (raw)

Initiation of convergence and extension movements of lateral mesoderm during zebrafish gastrulation

Developmental Dynamics, 2005

Embryonic morphogenesis is accomplished by cellular movements, rearrangements, and cell fate inductions. Vertebrate gastrulation entails morphogenetic processes that generate three germ layers, endoderm, mesoderm, and ectoderm, shaped into head, trunk, and tail. To understand how cell migration mechanistically contributes to tissue shaping during gastrulation, we examined migration of lateral mesoderm in the zebrafish. Our results illustrate that cell behaviors, different from mediolaterally oriented cell intercalation, also promote convergence and extension (C&E). During early gastrulation, upon internalization, individually migrating mesendodermal cells contribute to the elongation of the mesoderm by moving animally, without dorsal movement. Convergence toward dorsal starts later, by 70% epiboly (7.7 hpf). Depending on location along the Animal-Vegetal axis, an animal or vegetal bias is added to the dorsalward movement, so that paths fan out and the lateral mesoderm both converges and extends. Onset of convergence is independent of noncanonical Wnt signaling but is delayed when Stat3 signaling is compromised. To understand which aspects of motility are controlled by guidance cues, we measured turning behavior of lateral mesodermal cells. We show that cells exhibit directional preference, directionally-regulated speed, and turn toward dorsal when off-course. We estimate that ectoderm could supply from a fraction to all the dorsalward displacement seen in mesoderm cells. Using mathematical modeling, we demonstrate that directional preference is sufficient to account for mesoderm convergence and extension, and that, at minimum, two sources of guidance cues could orient cell paths realistically if located in the dorsal midline. Developmental Dynamics 234: 279 -292, 2005.

Cooperation of polarized cell intercalations drives convergence and extension of presomitic mesoderm during zebrafish gastrulation

Journal of Cell Biology, 2008

During vertebrate gastrulation, convergence and extension (C&E) movements narrow and lengthen the embryonic tissues, respectively. In zebrafish, regional differences of C&E movements have been observed; however, the underlying cell behaviors are poorly understood. Using time-lapse analyses and computational modeling, we demonstrate that C&E of the medial presomitic mesoderm is achieved by cooperation of planar and radial cell intercalations. Radial intercalations preferentially separate anterior and posterior neighbors to promote extension. In knypek;trilobite noncanonical Wnt mutants, the frequencies of cell intercalations are altered and the anteroposterior bias of radial intercalations is lost. This provides evidence for noncanonical Wnt signaling polarizing cell movements between different mesodermal cell layers. We further show using fluorescent fusion proteins that during dorsal mesoderm C&E, the noncanonical Wnt component Prickle localizes at the anterior cell edge, whereas D...

Regulation of Zebrafish Gastrulation Movements by slb/wnt11

2005

Epiboly movements Shortly after midblastula transition, the embryo consists of a mass of cells, the blastoderm, positioned on top of a big yolk cell; this stage is the so-called blastula stage. The blastoderm can be subdivided into an outer epithelium of enveloping layer (EVL) cells that cover the non-epithelial deep layer (DEL) cells, from which the embryo proper will form. Epiboly starts when EVL and DEL cells begin to spread over the yolk cell. In the DEL, epiboly is triggered by radial intercalations of cells deep within the blastula that move upwards into more superficial layers, thereby thinning the DEL along its 'innerouter' extent and expanding its coverage over the yolk cell (figure 1 A,B). The EVL layer, by contrast, does not undergo radial cell intercalations. Instead, it connects at its leading edge to the yolk cell membrane and moves towards the vegetal pole of the et al., 2004; Solnica-Krezel, 2005). Alternatively, since hypoblast cells simultaneously internalize around the circumference of the germ-ring, prechordal plate progenitor cells could be pushed away from the germring towards the animal pole by newly ingressing cells at the blastoderm margin rather than actively migrate into the anterior direction. This assumption is supported by studies showing that single wild-type cells transplanted into the germ-ring of mzoep mutantswhich lack nearly all mesendodermal induction and internalization-are able to internalize, but hardly move towards the animal pole, pointing at the possibility that this movement requires the simultaneous ingression of other mesendodermal cells (Carmany-Rampey and Schier, 2001; Montero et al., 2003). zebrafish schmalspur/sur mutants) and regulate the transcription of specific target genes. Among these genes are determinants of mesendodermal fates, like goosecoid (gsc) or no

Nodal signals mediate interactions between the extra-embryonic and embryonic tissues in zebrafish

Developmental Biology, 2007

In many vertebrates, extra-embryonic tissues are important signaling centers that induce and pattern the germ layers. In teleosts, the mechanism by which the extra-embryonic yolk syncytial layer (YSL) patterns the embryo is not understood. Although the Nodal-related protein Squint is expressed in the YSL, its role in this tissue is not known. We generated a series of stable transgenic lines with GFP under the control of squint genomic sequences. In all species, nodal-related genes induce their own expression through a positive feedback loop. We show that two tissue specific enhancers in the zebrafish squint gene mediate the response to Nodal signals. Expression in the blastomeres depends upon a conserved Nodal response element (NRE) in the squint first intron, while expression in the extra-embryonic enveloping layer (EVL) is mediated by an element upstream of the transcription start site. Targeted depletion experiments demonstrate that the zebrafish Nodal-related proteins Squint and Cyclops are required in the YSL for endoderm and head mesoderm formation. Thus, Nodal signals mediate interactions between embryonic and extra-embryonic tissues in zebrafish that maintain nodal-related gene expression in the margin. Our results demonstrate a high degree of functional conservation between the extra-embryonic tissues of mouse and zebrafish.

The Bmp Gradient of the Zebrafish Gastrula Guides Migrating Lateral Cells by Regulating Cell-Cell Adhesion

Current Biology, 2007

Background: Bone morphogenetic proteins (Bmps) are required for the specification of ventrolateral cell fates during embryonic dorsoventral patterning and for proper convergence and extension gastrulation movements, but the mechanisms underlying the latter role remained elusive. Results: Via bead implantations, we show that the Bmp gradient determines the direction of lateral mesodermal cell migration during dorsal convergence in the zebrafish gastrula. This effect is independent of its role during dorsoventral patterning and of noncanonical Wnt signaling. However, it requires Bmp signal transduction through Alk8 and Smad5 to negatively regulate Ca 2+ / Cadherin-dependent cell-cell adhesiveness. In vivo, converging mesodermal cells form lamellipodia that attach to adjacent cells. Bmp signaling diminishes the Cadherin-dependent stability of such contact points, thereby abrogating subsequent cell displacement during lamellipodial retraction. Conclusions: We propose that the ventral-to-dorsal Bmp gradient has an instructive role to establish a reverse gradient of cell-cell adhesiveness, thereby defining different migratory zones and directing lamellipodia-driven cell migrations during dorsal convergence in lateral regions of the zebrafish gastrula.

E-cadherin is required for gastrulation cell movements in zebrafish

Mechanisms of Development, 2005

E-cadherin is a member of the classical cadherin family and is known to be involved in cell-cell adhesion and the adhesion-dependent morphogenesis of various tissues. We isolated a zebrafish mutant (cdh1 rk3) that has a mutation in the e-cadherin/cdh1 gene. The mutation rk3 is a hypomorphic allele, and the homozygous mutant embryos displayed variable phenotypes in gastrulation and tissue morphogenesis. The most severely affected embryos displayed epiboly delay, decreased convergence and extension movements, and the dissociation of cells from the embryos, resulting in early embryonic lethality. The less severely affected embryos survived through the pharyngula stage and showed flattened anterior neural tissue, abnormal positioning and morphology of the hatching gland, scattered trigeminal ganglia, and aberrant axon bundles from the trigeminal ganglia. Maternal-zygotic cdh1 rk3 embryos displayed epiboly arrest during gastrulation, in which the enveloping layer (EVL) and the yolk syncytial layer but not the deep cells (DC) completed epiboly. A similar phenotype was observed in embryos that received antisense morpholino oligonucleotides (cdh1MO) against E-cadherin, and in zebrafish epiboly mutants. Complementation analysis with the zebrafish epiboly mutant weg suggested that cdh1 rk3 is allelic to half baked/weg. Immunohistochemistry with an anti-b-catenin antibody and electron microscopy revealed that adhesion between the DCs and the EVL was mostly disrupted but the adhesion between DCs was relatively unaffected in the MZcdh1 rk3 mutant and cdh1 morphant embryos. These data suggest that E-cadherin-mediated cell adhesion between the DC and EVL plays a role in the epiboly movement in zebrafish.

Zebrafish Nodal-Related 2encodes An Early Mesendodermal Inducer Signaling From the Extraembryonic Yolk Syncytial Layer

Developmental biology, 1998

Nodal-related factors have been implicated in mesodermal and neural patterning, and left-right asymmetry, in mouse, frog, and chicken embryos. We describe the isolation and characterization of zebrafish nodal-related 2 (znr2). znr2 is expressed at low levels maternally, and zygotic transcripts localize to dorsal blastomeres at MBT. Slightly later, znr2 is also expressed dorsally in the extraembryonic yolk syncytial layer (YSL). During early gastrulation, znr2 expression expands to include deep and superficial cells in the entire marginal zone and YSL. During shield stages, expression is primarily localized to superficial noninvoluting cells of the organizer called dorsal forerunners. Znr2 misexpression in whole fish embryos expands or duplicates dorsoanterior and axial cell fates. Furthermore, Znr2 overexpression exclusively in the YSL, a region implicated in endogenous mesodermal induction, causes broadened or duplicated gsc expression in the overlying blastoderm. Functional comparison of Znr2 and another recently identified zebrafish nodal-related factor, Znr1/Cyclops, reveals distinct inductive properties of each ligand. Znr2 efficiently induces organizer-type dorsoanterior mesodermal and endodermal markers, but only weakly, if at all, neural markers. In contrast, while Znr1/Cyclops reproducibly induces mesodermal and neural markers, it is an inefficient inducer of organizer-type mesoderm. Our results suggest that znr2 encodes a robust mesendodermal inducer that signals nonautonomously during the earliest stages of embryonic patterning, and that part of this activity arises from within the YSL.

Slb/Wnt11 controls hypoblast cell migration and morphogenesis at the onset of zebrafish gastrulation

Development, 2003

During vertebrate gastrulation, highly coordinated cellular rearrangements lead to the formation of the three germ layers, ectoderm, mesoderm and endoderm. In zebrafish, silberblick (slb)/wnt11 regulates normal gastrulation movements by activating a signalling pathway similar to the Frizzled-signalling pathway, which establishes epithelial planar cell polarity (PCP) in Drosophila. However, the cellular mechanisms by which slb/wnt11 functions during zebrafish gastrulation are still unclear. Using high-resolution two-photon confocal imaging followed by computer-assisted reconstruction and motion analysis, we have analysed the movement and morphology of individual cells in three dimensions during the course of gastrulation. We show that in slb-mutant embryos, hypoblast cells within the forming germ ring have slower, less directed migratory movements at the onset of gastrulation. These aberrant cell movements are accompanied by defects in the orientation of cellular processes along the individual movement directions of these cells. We conclude that slb/wnt11-mediated orientation of cellular processes plays a role in facilitating and stabilising movements of hypoblast cells in the germ ring, thereby pointing at a novel function of the slb/wnt11 signalling pathway for the regulation of migratory cell movements at early stages of gastrulation.

Cortical depth and differential transport of vegetally localized dorsal and germ line determinants in the zebrafish embryo

BioArchitecture, 2015

In zebrafish embryos, factors involved in both axis induction and primordial germ cell (PGC) development are localized to the vegetal pole of the egg. However, upon egg activation axis induction factors experience an asymmetric off-center shift whereas PGC factors undergo symmetric animally-directed movement. We examined the spatial relationship between the proposed dorsal genes wnt8a and grip2a and the PGC factor dazl at the vegetal cortex. We find that RNAs for these genes localize to different cortical depths, with the RNA for the PGC factor dazl at a deeper cortical level than those for axis-inducing factors. In addition, and in contrast to the role of microtubules in the long-range transport of dorsal determinants, we find that germ line determinant transport depends on the actin cytoskeleton. Our results support a model in which vegetal cortex differential RNA transport behavior is facilitated by RNA localization along cortical depth and differential coupling to cortical transport.