Hints from embryological studies guidance cues for. Germ cell determinant transmission, segregation, and. 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. Regulation of zebrafish primordial germ cell migration by attraction. 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. During their migration, zebrafish primordial germ cells pgcs rely on directional cues provided by the chemokine sdf1a, whose receptor is cxcr4b. Pgc migration in zebrafish takes place during the first 24 hours of its. Characterization of three novel members of the zebrafish pax258 family.
Yet, within 24 hours of development most of the cells arrive at distinct bilateral positions from which they coalesce into a gonad. Effects of specific and prolonged expression of zebrafish. In many lower animals, germ cell formation, migration and maintenance depend on maternally provided determinants in germ plasm. Cftr is required for the migration of primordial germ cells during. 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 cell development, maxplanckinstitute for biophysical chemistry, am fassberg 11, 37070 goettingen, germany.
Migration of primordial germ cells pgcs from their site of specification towards the. 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. Migration of primordial germ cells pgcs from their site of specification towards the developing gonad is controlled by directional cues from somatic tissues. To identify a molecular marker for the zebrafish pgcs, we cloned the zebrafish homologue of the drosophila vasa gene, which, in the fly. Three different egfp expression patterns were observed in f1 offspring obtained from different founder fish. The mechanism for primordial germcell migration is conserved. Active migration of zebrafish pgcs primordial germ cells.
Zebrafish germ cells contain granularlike structures, organized around the cell nucleus. Primordial germ cell migration in zebrafish wiley online library. Role of rho gtpases during primordial germ cell migration. The mechanism for primordial germcell migration is. Here, we used zebrafish model to investigate the role of cftr in primordial germ cells pgcs development. Studies have shown that overexpression of a dominantnegative form of the regulatory subunit of pi3k dnpi3k results in mis migration of zebrafish. Embryos lacking germ cells develop into sterile adult males.
Pgc migration in control zebrafish embryos by timelapse imaging. Generating chimeric zebrafish embryos by transplantation. Regulation of zebrafish primordial germ cell migration by attraction towards an. The molecular mechanisms whereby cxcr4b activation is interpreted intracellularly into directional migration are not known. Primordial germ cells pgcs give rise to gametes that are responsible for the development of a new organism in the next generation. 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. However, the involvement of cftr in regulating germ cell development remains largely unknown. Cftr is required for the migration of primordial germ cells during zebrafish early embryogenesis in reproduction authors. 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. 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. With permission from azadeh paksa, erez raz lab, muenster, germany. The orderly movement of a cell specialized to produce haploid.
B,c strong ubiquitous expression, d,e weak expression throughout the body, f,g strong expression in the brain and weak in the body. Zebrafish primordial germ cell migration embryology. Control over the morphology and segregation of zebrafish. 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. In the present study, we used statins, a class of potent competitive hmgcoar inhibitors. Correlative light and electron microscopy of rare cell. Home about cirm our publications grantee publications germ cell migration in zebrafish is cyclopaminesensitive but smoothenedindependent.
Signaling pathways controlling primordial germ cell. Cell migration matches nearly all research areas in cell and developmental biology, genetics, and biomedicine. Active migration of zebrafish pgcs primordial germ cells mehdi goudarzi. Cell migration in development and disease wiley online. Germ cells are labled in blue and nuclei of all cells are. Pgc migration in zebrafish is an excellent in vivo model system to study cell. Gcts occur in neonates, children, adolescents and young adults and can occur in the testis, the ovary or extragonadal sites. 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. 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.
Germ cell tumors gcts are malignant cancers that arise from embryonic precursors known as primordial germ cells. Specification of primordial germ cells in zebrafish. Chapters cover such topics as cell biology and developmental and neural biology. Maternal mir2025p is required for zebrafish primordial. 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. 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. B samples were embedded in 4% low melting point lmp agarose within a plastic mold. Germ cell migration in zebrafish is cyclopaminesensitive. Germ cell migration in zebrafish is dependent on hmgcoa. Figure 2migration of primordial germ cells is altered by atorvastatin treatment. These cells arise elsewhere and have to migrate across the embryo to reach developing gonadal precursors. 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 specification begins during cleavage in many animals or in the epiblast during gastrulation in birds and mammals. Primordial germ cells pgcs of zebrafish originate at random positions with respect to the dorsoventral axis. The field shows radical progress powered by the combination of new genomic tools, cell labeling techniques and the incorporation of new model systems. Specification of primordial germ cells in medaka oryzias. A zebrafish homologue of the chemokine receptor cxcr4 is a.
Weidinger g1, stebler j, slanchev k, dumstrei k, wise c, lovellbadge r, thisse c, thisse b, raz e. To determine the role of the germ line in somatic development of zebrafish, we ablated the pgcs by using dnd antisense mos. Additionally, cholesterol was found to be enriched in the genital ridges. 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. Germ cell determinant transmission, segregation, and function in. Regulation of zebrafish primordial germ cell migration by. 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.
After transport, involving passive movements and active migration, germ cells arrive at the developing gonads. Cells expressing germ plasm become primordial germ cells pgcs which will then give rise to the gametes. By continuing to use our website, you are agreeing to our use of cookies. A compact, yet separate animal and vegetal germ plasm aggregates at 16cell stage are found at the edge of the first two furrows. Zebrafish have emerged as a powerful model organism to study embryo morphogenesis.
Defect of which may impair germ cell development through elevation of. Interestingly, ifitm3 was also expressed in primordial germ cells pgcs from the bud stage to 24 hpf. Antitumor efficacy of a novel class of nonthiolcontaining peptidomimetic. The migration of zebrafish primordial germ cell towards the region where the gonad develops is guided by the chemokine sdf1a. 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. Little is known about the structure of these granules as well as their. Pgcs specification in zebrafish appears to depend on.
The embryo can be seen as a crescent shape at top of movie, pgcs can be seen as strongly labelled green cells. Primordial germ cells pgcs are segregated and specified from somatic cells during early development. In zebrafish, these p we use cookies to enhance your experience on our website. Pdf inducible sterilization of zebrafish by disruption. Twophoton microscopy of early germ cell migration in. In search of factors that affect the specification, maintenance or migration of primordial germ cells pgcs in zebrafish, we assayed embryos from a. Hypoxia impairs primordial germ cell migration in zebrafish danio. Primordial germ cells pgcs in zebrafish arise at distinct positions of the developing embryo and migrate towards their target guided by the chemokine sdf1a. 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. Cftr is required for the migration of primordial germ. Localizes to several cellular components, including the catenin complex.
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. The germ line development in mammals, on the other hand, occurs by induction and not by an endogenous germ plasm. Interestingly, in the mouse cholesterol synthesis seems to be involved in this process, as both cholesterol and isoprenoids are required to rescue this phenotype. The germ plasm effectively turns off gene expression to render the genome of the cell inert. Recent studies show that soon after their specification, the cells. 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.
A stepbystep guide to generating targeted chimeric zebrafish embryos by transplantation at the blastula or gastrula stage. 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. 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. Inducible sterilization of zebrafish by disruption of. In zebrafish, maternally inherited germ plasm ribonucleoparticles rnps have. However, since cell migration is a complicated process. Primordial germ cells pgcs are segregated and specified from somatic cells.
Inhibition of hmgcr in a tissue culture system impairs germ cell migration. 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. Primordial germ cell development in zebrafish sciencedirect. Although zebrafish has recently emerged as a model for vertebrate development, the primordial germ cells pgcs in this organism have not been previously described.
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