In mouse embryos, germ cells arise during gastrulation and migrate to the early gonad. First, they emerge from the primitive streak into the region of the endoderm that forms the hindgut. Later in ...development, a second phase of migration takes place in which they migrate out of the gut to the genital ridges. There, they co-assemble with somatic cells to form the gonad. In vitro studies in the mouse, and genetic studies in other organisms, suggest that at least part of this process is in response to secreted signals from other tissues. Recent genetic evidence in zebrafish has shown that the interaction between stromal cell-derived factor 1 (SDF1) and its G-protein-coupled receptor CXCR4, already known to control many types of normal and pathological cell migrations, is also required for the normal migration of primordial germ cells. We show that in the mouse, germ cell migration and survival requires the SDF1/CXCR4 interaction. First, migrating germ cells express CXCR4, whilst the body wall mesenchyme and genital ridges express the ligand SDF1. Second, the addition of exogenous SDF1 to living embryo cultures causes aberrant germ cell migration from the gut. Third, germ cells in embryos carrying targeted mutations in CXCR4 do not colonize the gonad normally. However, at earlier stages in the hindgut, germ cells are unaffected in CXCR4(-/-) embryos. Germ cell counts at different stages suggest that SDF1/CXCR4 interaction also mediates germ cell survival. These results show that the SDF1/CXCR4 interaction is specifically required for the colonization of the gonads by primordial germ cells, but not for earlier stages in germ cell migration. This demonstrates a high degree of evolutionary conservation of part of the mechanism, but also an area of evolutionary divergence.
Chemokines are vertebrate‐specific, structurally related proteins that function primarily in controlling cell movements by activating specific 7‐transmembrane receptors. Chemokines play critical ...roles in a large number of biological processes and are also involved in a range of pathological conditions. For these reasons, chemokines are at the focus of studies in developmental biology and of clinically oriented research aimed at controlling cancer, inflammation, and immunological diseases. The small size of the zebrafish embryos, their rapid external development, and optical properties as well as the large number of eggs and the fast expansion in genetic tools available make this model an extremely useful one for studying the function of chemokines and chemokine receptors in an in vivo setting. Here, we review the findings relevant to the role that chemokines play in the context of directed single‐cell migration, primarily in neutrophils and germ cells, and compare it to the collective cell migration of the zebrafish lateral line. We present the current knowledge concerning the formation of the chemokine gradient, its interpretation within the cell, and the molecular mechanisms underlying the cellular response to chemokine signals during directed migration.
Chemokines are vertebrate‐specific molecules with a wide range of activities in different aspects of animal physiology. Bussmann and Raz review here their involvement in single and collective cell migration in zebrafish.
The control over the acquisition of cell motility is central for a variety of biological processes in development, homeostasis, and disease. An attractive in vivo model for investigating the ...regulation of migration initiation is that of primordial germ cells (PGCs) in zebrafish embryos. In this study, we show that, following PGC specification, the cells can polarize but do not migrate before the time chemokine-encoded directional cues are established. We found that the regulator of G-protein signaling 14a protein, whose RNA is a newly identified germ plasm component, regulates the temporal relations between the appearance of the guidance molecules and the acquisition of cellular motility by regulating E-cadherin levels.
Single-cell migration is a key process in development, homeostasis, and disease. Nevertheless, the control over basic cellular mechanisms directing cells into motile behavior in vivo is largely ...unknown. Here, we report on the identification of a minimal set of parameters the regulation of which confers proper morphology and cell motility. Zebrafish primordial germ cells rendered immotile by knockdown of Dead end, a negative regulator of miRNA function, were used as a platform for identifying processes restoring motility. We have defined myosin contractility, cell adhesion, and cortex properties as factors whose proper regulation is sufficient for restoring cell migration of this cell type. Tight control over the level of these cellular features, achieved through a balance between miRNA-430 function and the action of the RNA-binding protein Dead end, effectively transforms immotile primordial germ cells into polarized cells that actively migrate relative to cells in their environment.
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► Zebrafish primordial germ cells (PGCs) depleted for Dead end (Dnd) are immotile ► Dnd-depleted PGCs are a platform for identifying processes promoting cell motility ► Dnd RNA binding function balance with miRNA action to control motility parameters ► Contractility, cortical rigidity, and cell adhesion modulation recover PGC motility
Using zebrafish primordial germ cells (PGCs) rendered immotile by knockdown of the RNA-binding protein Dnd, Goudarzi et al. define a combination of cellular parameters necessary and sufficient for bleb-based PGC motility. A balance between opposing actions of Dnd and miRNA-430 modulate these parameters in the control of cell motility.
Activation of the chemokine receptor CXCR4 by SDF1 controls a variety of biological processes in development, immune response, and disease 1–5. The carboxyl-terminal region of CXCR4 is subject to ...phosphorylation that allows binding of regulatory proteins 5; this results in downregulation of CXCR4 signaling and receptor internalization 6. Notably, truncations of this part of CXCR4 have been implicated in WHIM syndrome, a dominantly inherited immunodeficiency disorder 7, 8. Despite its importance in receptor signaling and the clinical relevance of its regulation, the precise function of regulating signaling level and internalization in controlling cell behavior is not known. Whereas a number of in vitro studies suggested that the carboxyl terminus of CXCR4 positively regulates chemotaxis (e.g., 9), others reached the opposite conclusion 8, 10, 11. These conflicting results highlight the importance of investigating this process under physiological conditions in the live animal. In this study, we demonstrate the significance of internalization and of controlling receptor signaling level for SDF-1-guided migration. We found that whereas internalization and the control over signaling intensity are dispensable for cell motility and directional sensing, they are essential for fine-tuning of migration in vivo, allowing precise arrival of zebrafish PGCs at their target, the region where the gonad develops.
The mechanisms that govern cell interactions during organ formation are not fully understood. In this issue of Developmental Cell, Miao et al. demonstrate a channel-independent role for gap junction ...proteins in the establishment of contacts between three cell types that build up the micropyle during oocyte development in Drosophila.
The mechanisms that govern cell interactions during organ formation are not fully understood. In this issue of Developmental Cell, Miao et al. demonstrate a channel-independent role for gap junction proteins in the establishment of contacts between three cell types that build up the micropyle during oocyte development in Drosophila.
During their migration, zebrafish primordial germ cells (PGCs) rely on directional cues provided by the chemokine SDF-1a, whose receptor is CXCR4b. The molecular mechanisms whereby CXCR4b activation ...is interpreted intracellularly into directional migration are not known. Here we investigate the role of two important biochemical pathways -- G-protein-dependent and phosphoinositide 3-kinase (PI3K)-dependent signaling -- in directing PGC migration in zebrafish. We show that G proteins of the Gi family are essential for directional migration but not for PGC motility. Inhibition of PI3K signaling in PGCs slows down their migration and leads to abnormal cell morphology as well as to reduced stability of filopodia. Invariably, during directed PGC migration, the distribution of the products of PI3K activity - phosphatidylinositol (3,4,5)-trisphosphate PtdIns(3,4,5)P(3) and/or phosphatidylinositol (3,4)bisphosphate PtdIns(3,4)P(2) -- is not polarized, and reducing the level of these 3-phosphoinositides does not affect the ability of PGCs to migrate directionally. We therefore conclude that Gi-dependent signaling is essential for directional migration, whereas the PI3K pathway is important for the actual motility of PGCs.
As in many other animals, the primordial germ cells (PGCs) in avian and reptile embryos are specified in positions distinct from the positions where they differentiate into sperm and egg. Unlike in ...other organism however, in these embryos, the PGCs use the vascular system as a vehicle to transport them to the region of the gonad where they exit the blood vessels and reach their target. To determine the molecular mechanisms governing PGC migration in these species, we have investigated the role of the chemokine stromal cell-derived factor-1 (SDF-1/CXCL12) in guiding the cells towards their target in the chick embryo. We show that sdf-1 mRNA is expressed in locations where PGCs are found and towards which they migrate at the time they leave the blood vessels. Ectopically expressed chicken SDF-1alpha led to accumulation of PGCs at those positions. This analysis, as well as analysis of gene expression and PGC behavior in the mouse embryo, suggest that in both organisms, SDF-1 functions during the second phase of PGC migration, and not at earlier phases. These findings suggest that SDF-1 is required for the PGCs to execute the final migration steps as they transmigrate through the blood vessel endothelium of the chick or the gut epithelium of the mouse.
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