Dorsal-ventral Polarity Research Articles

The Drosophila TGF-α-like protein Gurken: expression and cellular localization during Drosophila oogenesis

The establishment of anterior-posterior and dorsal-ventral polarity of the Drosophila egg and embryo depends on the function of the genes gurken, cornichon and Egfr ( Drosophila epidermal growth factor receptor homolog). These genes encode components of a signal transduction pathway that transmits information between the germline cells and the somatic follicle cells of the ovary, gurken encodes a transforming growth factor-α-like protein and is a putative germline ligand of the Egfr present on the follicle cells. In mid-oogenesis the gurken transcript becomes spatially localized to the future dorsal-anterior cortex of the oocyte. To analyze the distribution pattern of Gurken protein we prepared antibodies against Gurken. We describe here the distribution pattern of the Gurken protein in wild-type ovaries and in ovaries from a number of dorsal-ventral patterning mutants. By immunoblotting we detect one major form of the Gurken protein, which likely corresponds to the unprocessed protein.

Restricting oxygen supply to the prospective dorsal side does not reverse axis polarity in embryos of Xenopus laevis

During the first cell cycle, the prospective dorsal side of the embryo of Xenopus laevis becomes enriched in mitochondria relative to the ventral side. This differential distribution of mitochondria persists throughout early development, but it is not known if it is of functional significance, since there do not appear to be dorsal-ventral differences in metabolic rate. However, the unilateral anaerobiosis experiments of Landström and Løvtrup do suggest a role for energy metabolism in determining axis polarity. These experiments apparently show that restricting oxygen supply to the prospective dorsal side causes a reversal of dorsal-ventral axis polarity. We have reinvestigated this point using cell-marking techniques. We find that although gastrulation is initiated at the open end of the tube, the polarity of neural plate development is unaffected. Thus, definitive dorsal-ventral polarity is not affected by the experimental treatment, and it is unlikely that gradients of energy metabolism have a role in specifying axis polarity in X. laevis.

BrainiacEncodes a Novel, Putative Secreted Protein That Cooperates with Grk TGFα in the Genesis of the Follicular Epithelium

brainiac (brn)is involved in a number of developmental events. In addition to being required zygotically for segregation of neuroblasts from epidermoblasts, it is essential for a series of critical steps during oogenesis which also depend upongurken (grk),a TGFα homolog. Animals harboring strong mutations of eithergrkor EGF receptor tyrosine kinase (Egfr) or doubly mutant forbrnand weakgrkorEgfrmutations produce ovarian follicles with multiple sets of nurse cell–oocyte complexes. These follicles frequently have discontinuities in the follicular epithelium that uncover nurse cells but not the oocyte. Gaps first appear in the germarium, suggesting that some nurse cells lack affinity for invading prefollicular cells. This is the first evidence thatgrk,in addition to its involvement in the genesis of anterior–posterior and dorsal–ventral polarity, is also required for Egfr-dependent development of the follicular epithelium that surrounds each nurse cell/oocyte cluster to form an egg chamber. We have used restriction fragment length polymorphisms to localizebrnto a 10-kb region within a 300-kb stretch of DNA on the X-chromosome, and we have identified thebrngene by means of RNA rescue.brncodes for a putative secreted protein.brnis expressed in germ cells at the time follicle cells first surround the nurse cell–oocyte complex. Our genetic data suggest thatbrnacts in a parallel, but partially overlapping pathway to the Grk–Egfr signaling pathway. Thebrnpathway may help to provide specificity to TGFα–Egfr function during oogenesis.

Intercellular signaling and the polarization of body axes during Drosophila oogenesis.

The anterior-posterior and dorsal-ventral axes of the Diosophila egg and embryo are established during oogen­ esis as the egg is being formed. The mechanisms under­ lying this process involve intercellular signaling events, including bidirectional communication between germline and somatic cells, and local cell-cell interactions in the soma. On the molecular level, these interactions ap­ pear to be mediated by a small number of intercellular signaling systems, including the Epidermal growth fac­ tor receptor [Egfr] and Notch systems, that are used mul­ tiple times during oogenesis to trigger different develop­ mental switches. The precisely regulated interplay be­ tween these various signaling systems forms a network of interdependencies that leads to the establishment of both the anterior-posterior and dorsal-ventral polarity in the egg chamber and embryo. Because both the Egfr and Notch signaling systems are widely conserved, the mechanisms involved in the regulation of these pro­ cesses in Drosophila oogenesis may serve as a paradigm for understanding intercellular signaling events in other organisms.

β-catenin translocation into nuclei demarcates the dorsalizing centers in frog and fish embryos

The question of how dorsal-ventral polarity is established in vertebrates is central to our understanding of their early development. Several lines of evidence suggest that wnt-signaling is involved in the induction of dorsal-specific gene expression in the Spemann Organizer of amphibians. Here, we show that β-catenin, acting as a component of the wnt-pathway, transiently accumulates in nuclei on the dorsal side of Xenopus and zebrafish blastulae. The spatially restricted nuclear translocation of β-catenin precedes the expression of dorsal-specific genes. In experimentally ventralized frog embryos the dorsal-ventral pattern of β-catenin nuclear staining is abolished; in contrast, embryos hyperdorsalized by Li-ions or by injection of Xwnt8-mRNA exhibit an enhanced nuclear accumulation of β-catenin. The results show that translocation of β-catenin into nuclei in the wake of wnt-signaling is an early step in the establishment of the dorsal-ventral axis in frog and fish embryos.

Functional interactions between the pelle kinase, Toll receptor, and tube suggest a mechanism for activation of dorsal.

A complex signal transduction pathway functions in the early Drosophila embryo to establish dorsal-ventral polarity. Activation of this pathway results in the nuclear transport of the protein dorsal (dl), a member of the rel/NF-kappaB family of transcription factors. Genetic studies have identified three intracellular components whose activity is required for activation of dl: Toll, a transmembrane receptor; pelle (pll), a serine/threonine protein kinase; and tube, a protein of unknown function. Here we examine the activities of these proteins when coexpressed in Drosophila Schneider cells. Coexpression of pll with dl enhanced dl nuclear localization and resulted in a modest increase in transcriptional activity. However, when pll was coexpressed with a specific mutant derivative of Toll (TlNaeI), although not with wild-type Toll, a striking synergistic activation of dl was detected. Unexpectedly, coexpression of pll plus TlNaeI, in the absence of dl, resulted in a similar synergistic activation of a GAL4-tube fusion protein. Based on these and other results, we propose a model in which pll receives a signal from activated Toll and phosphorylates tube, which then participates directly in dl activation.

Multiple signaling pathways establish both the individuation and the polarity of the oocyte follicle in Drosophila.

The development of the Drosophila oocyte depends upon a sequential series of interactions between the germline cells and the somatically derived follicle cells to produce individual follicles with appropriate polarities. In the germarium the control of germline cell division depends upon a proper interaction with somatic cells adjacent to the germline stem cells. Both gurken and brainiac are required in the germline, and the Egfr, daughterless, Notch, and Delta genes are required in the somatic cells to produce individual egg chambers with a continuous follicular epithelium. After a follicle forms, components in these same signaling pathways, plus additional genes, are then required for the establishment of the anterior-posterior polarity, followed by the dorsal-ventral polarity of the developing follicle. Initially, gurken mRNA is localized to the posterior edge of the oocyte, where it signals the posterior polar follicle cells to differentiate as posterior. The anterior-posterior assymmetry of the oocyte is then established by a reorganization of the microtubule network, which require a Notch-Delta-dependent signal sent from the posterior polar follicle cells to the oocyte and the activity of protein kinase A in the germ line. This reorganization leads to the localization of the maternal anterior-posterior determinants bicoid and oskar to opposite poles of the oocyte and the repositioning of the oocyte nucleus to the anterior-dorsal surface of the oocyte, gurken mRNA and protein are now concentrated between the oocyte nucleus and the adjacent anterior-dorsal follicle cells, where, in combination with Rhomboid, it locally activates the EGF receptor and its downstream cascade to direct the adjoining cells to adopt a dorsal fate. This process is thought to restrict the action of three follicle cell gene functions, encoded by windbeutel, nudal, and, pipe, to the ventral follicle cells, where they lead to the localized activation of a serine protease cascade required to produce the active Spätzle ligand to activate the Toll receptor. Finally, the termini of the embryo are dependent upon the activation of the Torso receptor, and this requires the localized expression of torso-like in a subset of follicle cells at the anterior and posterior poles of the follicle, which leads to the activation of Trunk, the putative ligand for Torso. In summary, the normal development of the oocyte requires a continuous sequence of germline-follicle cell interactions to provide the polarities responsible for normal development.

Pattern formation: The link between ovary and embryo.

Abstract The Drosophila gene nudel may encode a spatially restricted serine protease involved in producing the ligand for the receptor Toll and linking dorsal–ventral polarity in the egg chamber to the developing embryonic axis.

Spatial regulation of Drosophila snake protease activity in the generation of dorsal-ventral polarity.

Positional information along the dorsal-ventral axis of the Drosophila embryo is acquired through a signal transduction pathway which employs a extracellular protease cascade. The sequential activation of serine protease zymogens results in the ventrally localized production of a ligand in the perivitelline space of the embryo. Snake is one of several serine proteases which function in generating the ventralizing signal. Here, we investigate the biochemical properties of Snake in vivo and in vitro using recombinant forms of the protease. Wild-type Snake zymogen completely rescues embryos from snake null females when microinjected into the perivitelline space. Biochemical evidence for a covalently associated two-chain form of the activated protease is presented. The contribution of the activation peptide region to zymogen activation was addressed using site-directed mutagenesis. The phenotypic rescue properties of an autoactivated form of Snake reveal that the covalently associated proenzyme polypeptide chain suppresses a dominant effect associated with the activated catalytic chain alone. Recombinant active catalytic chain was produced and found to be short lived as a recombinant protein. These results suggest a model in which the proenzyme polypeptide both stabilizes and targets the Snake catalytic chain to a ventrally localized activation complex within the perivitelline space.

Strategies for the generation of neuronal diversity in the developing central nervous system

During development, the neural tube produces a large diversity of neuronal phenotypes from a morphologically homogeneous pool of precursor cells. In recent years, the cellular and molecular mechanisms by which specific types of neurons are generated have been explored, in the hope of discovering features common to development throughout the nervous system. This article focuses on three strategies employed by the CNS to generate distinct classes of neuronal phenotypes during development: dorsal-ventral polarization in the spinal cord, segmentation in the hindbrain, and a lamination in the cerebral cortex. The mechanisms for neurogenesis exemplified by these three strategies range from a relatively rigid, cell lineage-dependent specification with a high degree of subservance to early patterns of gene expression, to inductions and cell-cell interactions that determine cell fates more flexibly.

Pattern formation. Gurken meets torpedo for the first time.

Intercellular communication between oocyte and follicle cells, mediated by the gurken-torpedo/DER signalling pathway, has a crucial role in determining both anterior-posterior and dorsal-ventral polarity in Drosophila.

The anterior-posterior and dorsal-ventral axes have a common origin in Drosophila melanogaster.

The mechanisms governing anterior-posterior and dorsal-ventral polarity in Drosophila melanogaster had previously been considered as independent processes. However, two papers(1,2) now reveal that both axes are initiated during oogenesis by the same pathway, and also clearly demonstrate that one is dependent on the other.

Bone morphogenetic protein acts as a ventral mesoderm modifier in early Xenopus embryos.

Mesoderm of early vertebrate embryos gradually acquires dorsal-ventral polarity during embryogenesis. This specification of mesoderm is thought to be regulated by several polypeptide growth factors. Bone morphogenetic protein (BMP), a member of the TGF-β family, is one of the regulators suggested to be involved in the formation of ventral mesoderm. In this paper, the nature of the endogenous BMP signal in dorsal-ventral specification was assessed in early Xenopus embryos using a dominant negative mutant of the Xenopus BMP receptor. In ectodermal explant assays, disruption of endogenous BMP signaling by the mutant receptor changed the competence of the explant cells to mesoderm-inducing factors, activin and basic fibroblast growth factor (bFGF), and led to formation of neural tissue without mesoderm induction. This result suggests that endogenous BMP acts as a ventral mesoderm modifier rather than a ventral mesoderm inducer, and that interactions between endogenous BMP and mesoderm-inducing factors may be important in dorsal-ventral patterning of embryonic mesoderm. In addition, the induction of neural tissue by inhibition of the BMP signaling pathway also suggests involvement of BMP in neural induction.

How many signals does it take?

Although the genetics of dorsal-ventral polarity which leads to mesoderm formation in Drosophila are understood in considerable detail, subsequent molecular mechanisms involved in patterning the mesoderm primordium into individual mesodermal subtypes are poorly understood. Two papers published recently suggest strongly that an inductive signal from dorsal ectoderm is involved in subdividing the underlying mesoderm, and present evidence that one of the signalling factors is Decapentaplegic (Dpp), a member of the bone morphogenetic protein subgroup of the Transforming Growth Factor-beta (TGF-beta) super family of proteins.

Cornichon and the EGF receptor signaling process are necessary for both anterior-posterior and dorsal-ventral pattern formation in Drosophila

In Drosophila, the dorsal-ventral polarity of the egg chamber depends on the localization of the oocyte nucleus and the gurken RNA to the dorsal-anterior corner of the oocyte. Gurken protein presumably acts as a ligand for the Drosophila EGF receptor ( torpedo/DER) expressed in the somatic follicle cells surrounding the oocyte. cornichon is a gene required in the germline for dorsal-ventral signaling. cornichon, gurken, and torpedo also function in an earlier signaling event that establishes posterior follicle cell fates and specifies the anterior-posterior polarity of the egg chamber. Mutations in all three genes prevent the formation of a correctly polarized microtubule cytoskeleton required for proper localization of the anterior and posterior determinants bicoid and oskar and for the asymmetric positioning of the oocyte nucleus.

Multiple cis-acting targeting sequences are required for orb mRNA localization during Drosophila oogenesis.

The targeting of positional information to specific regions of the oocyte or early embryo is one of the key processes in establishing anterior-posterior and dorsal-ventral polarity. In many developmental systems, this is accomplished by localization of mRNAs. The germ line-specific Drosophila orb gene plays a critical role in defining both axes of the developing oocyte, and its mRNA is localized in a complex pattern during oogenesis. We have identified a 280-bp sequence from the orb 3' untranslated region capable of reproducing this complex localization pattern. Furthermore, we have found that multiple cis-acting elements appear to be required for proper targeting of orb mRNA.

Multiple cis-acting targeting sequences are required for orb mRNA localization during Drosophila oogenesis.

The targeting of positional information to specific regions of the oocyte or early embryo is one of the key processes in establishing anterior-posterior and dorsal-ventral polarity. In many developmental systems, this is accomplished by localization of mRNAs. The germ line-specific Drosophila orb gene plays a critical role in defining both axes of the developing oocyte, and its mRNA is localized in a complex pattern during oogenesis. We have identified a 280-bp sequence from the orb 3' untranslated region capable of reproducing this complex localization pattern. Furthermore, we have found that multiple cis-acting elements appear to be required for proper targeting of orb mRNA.

Cell intercalation during Drosophila germband extension and its regulation by pair-rule segmentation genes

After the onset of gastrulation, the Drosophila germband undergoes a morphological change in which its length along the anterior-posterior axis increases over two-and-a-half fold while its width along the dorsal-ventral axis simultaneously narrows. The behavior of individual cells during germband extension was investigated by epi-illumination and time-lapse video microscopy of living embryos. Cells intercalate between their dorsal and ventral neighbors during extension, increasing the number of cells along the anterior-posterior axis while decreasing the number of cells along the dorsal-ventral axis. Mutations that reduce segmental subdivision of the embryo along the anterior-posterior axis decrease both germband extension and its associated cell intercalation. In contrast, cell intercalation and germband extension are still detected in embryos that lack dorsal-ventral polarity. Characterization of germband extension and cell intercalation in mutant embryos with altered segmentation gene expression indicates that these processes are regionally autonomous and are dependent upon the establishment of striped expression patterns for certain pair-rule genes. Based on these observations, we propose a model for germband extension in which cell intercalation results from the establishment of adhesive differences between stripes of cells by pair-rule genes.

Ventralizing signal determined by protease activation in Drosophila embryogenesis.

Specification of dorsal-ventral cell fate during Drosophila embryogenesis is mediated by a signal transduction pathway. Asymmetry of cell fates arises through the spatially restricted production of a ligand in an extracellular compartment called the perivitelline space. The snake and easter genes are required for the production of the ligand and they encode the proenzyme form of secreted extracellular serine proteases. We have examined the effect of producing a preactivated form of the snake protease on the generation of dorsal-ventral polarity. SP6 RNA microinjection experiments reveal that different cell fates acquired at cellular blastoderm can be specified by the amount and spatial distribution of activated snake protein. Our results support a protease cascade model in which localized activation of uniformly distributed protease proenzymes leads to the spatially restricted production of ligand in the perivitelline space on the ventral side of the embryo.

The maternal genes apx-1 and glp-1 and establishment of dorsal-ventral polarity in the early C. elegans embryo

The sister biastomeres ABp and ABa are equipotent at the beginning of the 4-cell stage in C. elegans embryos, but soon become committed to different fates. We show that the glp-1 gene, a homolog of the Notch gene of Drosophila, functions in two distinct cell-cell interactions that specify the ABp and ABa fates. These interactions both require maternal expression of glp-1. We show that a second maternal gene, apx-1, functions with glp-1 only in the specification of the ABp fate and that apx-1 can encode a protein homologous to the Delta protein of Drosophila. Our results suggest how interactions mediated by glp-1 and apx-1 contribute to the establishment of the dorsal-ventral axis in the early C. elegans embryo.