Pdf inducible sterilization of zebrafish by disruption. Cftr is required for the migration of primordial germ cells during. In many lower animals, germ cell formation, migration and maintenance depend on maternally provided determinants in germ plasm. During zebrafish development, a gradient of stromalderived factor 1a sdf1a provides the directional cue that guides the migration of the primordial germ cells pgcs to the gonadal tissue. The mechanism for primordial germcell migration is conserved. To determine the role of the germ line in somatic development of zebrafish, we ablated the pgcs by using dnd antisense mos. Here we describe a method to produce large numbers of infertile fish by inducing ubiquitous expression of sdf1a in zebrafish embryos resulting in disruption of the normal pgc migration pattern. New insights into the control of these remaining mysteries have emerged from genetic dissection and live imaging of germ cell migration in drosophila, zebrafish, and mouse embryos. Gcts occur in neonates, children, adolescents and young adults and can occur in the testis, the ovary or extragonadal sites. Cftr is required for the migration of primordial germ cells during zebrafish early embryogenesis mutations in the cystic fibrosis transmembrane conductance regulator cftr gene affect fertility in both sexes.
A compact, yet separate animal and vegetal germ plasm aggregates at 16cell stage are found at the edge of the first two furrows. To identify a molecular marker for the zebrafish pgcs, we cloned the zebrafish homologue of the drosophila vasa gene, which, in the fly. The mechanism for primordial germcell migration is. Studies have shown that overexpression of a dominantnegative form of the regulatory subunit of pi3k dnpi3k results in mis migration of zebrafish. Regulation of zebrafish primordial germ cell migration by.
Specification of primordial germ cells in medaka oryzias. Migration of primordial germ cells pgcs from their site of specification towards the developing gonad is controlled by directional cues from somatic tissues. When expression of cxcr4b or its ligand is inhibited, the migration of germ cells is disrupted, indicating that sdf1a acts as a potent chemoattractant for zebrafish pgcs. Germ cell development, maxplanckinstitute for biophysical chemistry, am fassberg 11, 37070 goettingen, germany. Characterization of three novel members of the zebrafish pax258 family. Recent studies show that soon after their specification, the cells. Role of rho gtpases during primordial germ cell migration. Germ plasm to primordial germ cell transition in zebrafish. B samples were embedded in 4% low melting point lmp agarose within a plastic mold. Embryos lacking germ cells develop into sterile adult males. The zebrafish is an excellent model for in vivo studies of vertebrate germ line because of the availability of molecular markers and the transparency of developing zebrafish embryos, which permits direct visualization of pgc development, a coordinated sequence of events involving cell fate specification, proliferation, and directed migration.
Maternal mir2025p is required for zebrafish primordial. Primordial germ cell development in zebrafish sciencedirect. Localizes to several cellular components, including the catenin complex. Pgc migration in zebrafish is an excellent in vivo model system to study cell. Primordial germ cells pgcs in zebrafish arise at distinct positions of the developing embryo and migrate towards their target guided by the chemokine sdf1a. Specification of primordial germ cells in zebrafish. Cells of interest in red primordial germ cells pgcs in this example, marked with asterisk. Germ cell specification begins during cleavage in many animals or in the epiblast during gastrulation in birds and mammals. However, since cell migration is a complicated process.
Due to their optical clarity, they are uniquely suited for timelapse imaging studies, providing insights into the dynamic processes underlying tissue formation and cell migration. These cells follow a characteristic developmental path that is manifested in specialized regulation of basic cell functions and behavior making them an attractive system for studying cell fate specification, differentiation and migration. Interestingly, in the mouse cholesterol synthesis seems to be involved in this process, as both cholesterol and isoprenoids are required to rescue this phenotype. During their migration, zebrafish primordial germ cells pgcs rely on directional cues provided by the chemokine sdf1a, whose receptor is cxcr4b. Here we apply genetics and in vivo imaging to show that odysseus, a zebrafish homologue of the gproteincoupled chemokine receptor cxcr4, is required specifically in germ cells for their. Although the gcl homologue genes have been identified in some organisms, there is little data on the expression pattern and functional analysis of the gcl gene in zebrafish. In zebrafish, these p we use cookies to enhance your experience on our website. Although zebrafish has recently emerged as a model for vertebrate development, the primordial germ cells pgcs in this organism have not been previously described. The orderly movement of a cell specialized to produce haploid. These structures share common features with polar granules in drosophila, germinal granules in xenopus and chromatoid bodies in mice germ cells, such as the localization of the zebrafish vasa, piwi and nanos proteins, among others. Pgcs specification in zebrafish appears to depend on.
Cell migration in development and disease wiley online. Inhibition of hmgcr in a tissue culture system impairs germ cell migration. Antitumor efficacy of a novel class of nonthiolcontaining peptidomimetic. Pgc migration in zebrafish takes place during the first 24 hours of its. Germ cell migration in zebrafish is cyclopaminesensitive. Cell migration matches nearly all research areas in cell and developmental biology, genetics, and biomedicine. These cells arise elsewhere and have to migrate across the embryo to reach developing gonadal precursors. Pgc migration in control zebrafish embryos by timelapse imaging. Home about cirm our publications grantee publications germ cell migration in zebrafish is cyclopaminesensitive but smoothenedindependent. In zebrafish, maternally inherited germ plasm ribonucleoparticles rnps have. Zebrafish germ cells contain granularlike structures, organized around the cell nucleus. Hypoxia impairs primordial germ cell migration in zebrafish danio. After transport, involving passive movements and active migration, germ cells arrive at the developing gonads. A stepbystep guide to generating targeted chimeric zebrafish embryos by transplantation at the blastula or gastrula stage.
Cellular and developmental biology, part b developmental biology, the second volume on the topic in the methods in cell biology series, looks at methods for analyzing cellular and developmental biology of zebrafish. Regulation of zebrafish primordial germ cell migration by attraction. Additionally, cholesterol was found to be enriched in the genital ridges. Correlative light and electron microscopy of rare cell. Little is known about the structure of these granules as well as their. Primordial germ cells pgcs of zebrafish originate at random positions with respect to the dorsoventral axis. By continuing to use our website, you are agreeing to our use of cookies. The germ plasm effectively turns off gene expression to render the genome of the cell inert.
Inducible sterilization of zebrafish by disruption of. The field shows radical progress powered by the combination of new genomic tools, cell labeling techniques and the incorporation of new model systems. With permission from azadeh paksa, erez raz lab, muenster, germany. In sexually reproducing organisms, primordial germ cells pgcs give rise to gametes that are responsible for the development of a new organism in the next generation. Figure 2migration of primordial germ cells is altered by atorvastatin treatment. Twophoton microscopy of early germ cell migration in. Chapters cover such topics as cell biology and developmental and neural biology. The migration of zebrafish primordial germ cell towards the region where the gonad develops is guided by the chemokine sdf1a. Migration of primordial germ cells pgcs from their site of specification towards the. The zebrafish has become an excellent model system for studying the mechanisms of germ cell development and function.
A zebrafish homologue of the chemokine receptor cxcr4 is a. Active migration of zebrafish pgcs primordial germ cells mehdi goudarzi. Control over the morphology and segregation of zebrafish. In search of factors that affect the specification, maintenance or migration of primordial germ cells pgcs in zebrafish, we assayed embryos from a. Primordial germ cell migration in zebrafish wiley online library. Cells expressing germ plasm become primordial germ cells pgcs which will then give rise to the gametes. Although in several animals the pgcs are attracted by signals emanating from their final target, the gonadal mesoderm, little is known about the mechanisms that control earlier steps of migration. The molecular mechanisms whereby cxcr4b activation is interpreted intracellularly into directional migration are not known. Hints from embryological studies guidance cues for. Active migration of zebrafish pgcs primordial germ cells. Yet, within 24 hours of development most of the cells arrive at distinct bilateral positions from which they coalesce into a gonad. Two modes of germ line specification have been described. Insulinlike growth factors igfs and their receptors, which can function by activation of the pi3k pathway, are also required for zebrafish primordial germ cell migration and survival 7.
Germ cells are labled in blue and nuclei of all cells are. Regulation of zebrafish primordial germ cell migration by attraction towards an. The embryo can be seen as a crescent shape at top of movie, pgcs can be seen as strongly labelled green cells. Cftr is required for the migration of primordial germ cells during zebrafish early embryogenesis in reproduction authors. Zebrafish dead end rna is a component of the germ plasm and is specifically expressed in primordial germ cells in a largescale wholemount in situ screen for genes expressed in zebrafish pgcs, we identified a novel gene, dead end dnd, which is specifically expressed in germ plasm and primordial germ cells. Generating chimeric zebrafish embryos by transplantation. Defect of which may impair germ cell development through elevation of. In the present study, we used statins, a class of potent competitive hmgcoar inhibitors. However, the involvement of cftr in regulating germ cell development remains largely unknown. The germ line development in mammals, on the other hand, occurs by induction and not by an endogenous germ plasm. Cftr is required for the migration of primordial germ.
Primordial germ cells pgcs are the progenitors of reproductive cells in metazoans and are an important model for the study of cell migration in vivo. Weidinger g1, stebler j, slanchev k, dumstrei k, wise c, lovellbadge r, thisse c, thisse b, raz e. Germ cell migration in zebrafish is dependent on hmgcoa. Germcell specification in fish was studied mainly in bony fish teleosts where the pgcs were recognized by their large size and by the typical electron dense nuage structures that are found in germ cells of different organisms. Effects of specific and prolonged expression of zebrafish. Zebrafish primordial germ cell migration embryology.
Primordial germ cells pgcs give rise to gametes that are responsible for the development of a new organism in the next generation. Germ cell tumors gcts are malignant cancers that arise from embryonic precursors known as primordial germ cells. This expression analysis indicates that zebrafish ifitm1 may play a critical role in early organogenesis and may perform immune or hematopoietic functions and ifitm3 might be necessary for pgc migration and the formation of female germ cells. Germ cell determinant transmission, segregation, and. Zebrafish embryos develop rapidly and externally so early aspects of germ cell development, such as primordial germ cell specification and migration, can be observed and manipulated in living animals raz and reichman. Germ cell determinant transmission, segregation, and function in. Three different egfp expression patterns were observed in f1 offspring obtained from different founder fish. Read germ cell migration in zebrafish is cyclopaminesensitive but smoothenedindependent, developmental biology on deepdyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips. Primordial germ cells pgcs are segregated and specified from somatic cells. Primordial germ cells pgcs are segregated and specified from somatic cells during early development. As described 29, 30, dnd is essential for normal migration and survival of pgcs, and therefore, embryos devoid of this protein develop to become sterile adults. Germ cell migration in zebrafish is dependent on hmgcoa reductase activity and. Huijuan liao 1, yan chen 1, yulong li 1, 2, shaolong xue 1, 3, mingfeng liu 1, 4, ziyuan lin 1, yanyan liu 5, hsiao chang chan 1, 3, xiaohu zhang 1 and huaqin sun 1. Here, we used zebrafish model to investigate the role of cftr in primordial germ cells pgcs development.
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