RESUMO
Craniofrontonasal syndrome (CFNS) is an X-linked disorder caused by inactivating mutations in the gene for ephrin-B1 (EFNB1). Paradoxically it shows a more severe phenotype in females than in males. As a result of X inactivation cell populations with and without EFNB1 expression are found in EFNB1+/- females. This is thought to initiate a process termed cellular interference which may be responsible for the phenotype in females. We present a boy with severe clinical features of CFNS. In â¼42% of his blood cells we found a supernumerary ring X chromosome containing EFNB1 but lacking XIST. Mosaicism for cell populations with different levels of EFNB1 expression can explain the severe phenotype of this patient. In vitro experiments in Xenopus tissue showed that cells overexpress ephrinB1 cluster and sort out from wild-type cells. Our report provides further evidence that cellular interference contributes to the paradoxical inheritance pattern of CFNS.
Assuntos
Anormalidades Craniofaciais/genética , Efrina-B1/genética , Animais , Cromossomos Humanos X , Efrina-B1/metabolismo , Humanos , Lactente , Masculino , Mosaicismo , RNA Longo não Codificante/genética , Síndrome , Xenopus/genéticaRESUMO
Cell signalling through Frizzled receptors has evolved to considerable complexity within the metazoans. The Frizzled-dependent signalling cascade comprises several branches, whose differential activation depends on specific Wnt ligands, Frizzled receptor isoforms and the cellular context. In Xenopus laevis embryos, the canonical beta-catenin pathway contributes to the establishment of the dorsal-ventral axis. A different branch, referred to as the planar cell polarity pathway, is essential for cell polarization during elongation of the axial mesoderm by convergent extension. Here we demonstrate that a third branch of the cascade is independent of Dishevelled function and involves signalling through trimeric G proteins and protein kinase C (PKC). During gastrulation, Frizzled-7 (Fz7)-dependent PKC signalling controls cell-sorting behaviour in the mesoderm. Loss of zygotic Fz7 function results in the inability of involuted anterior mesoderm to separate from the ectoderm, which leads to severe gastrulation defects. This result provides a developmentally relevant in vivo function for the Fz/PKC pathway in vertebrates.
Assuntos
Gástrula/fisiologia , Receptores de Superfície Celular/fisiologia , Receptores Acoplados a Proteínas G , Transdução de Sinais , Transativadores , Proteínas de Xenopus , Animais , Polaridade Celular , Proteínas do Citoesqueleto/metabolismo , Embrião não Mamífero , Fatores de Crescimento de Fibroblastos/farmacologia , Proteínas de Ligação ao GTP/metabolismo , Gástrula/citologia , Gástrula/metabolismo , Expressão Gênica/efeitos dos fármacos , Mesoderma/fisiologia , Morfogênese/fisiologia , Oligonucleotídeos Antissenso/farmacologia , Proteína Quinase C/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Xenopus laevis , beta CateninaRESUMO
The members of the Pitx family of homeobox transcription factors have been involved in many aspects of vertebrate embryogenesis, like for example, development of teeth, eyes and limbs. We previously reported expression patterns and function of Pitx2c in the generation of laterality and asymmetric morphogenesis of heart and gastro-intestinal tract in mouse, frog and zebrafish (Development 126 (1999) 1225; Mech. Dev. 90 (2000) 41). Here we describe the differential expression of Pitx1, Pitx2b and Pitx2c during anterior ectodermal pattern formation and differentiation of cement gland, stomodeum and pituitary in the frog Xenopus laevis.
Assuntos
Embrião não Mamífero/metabolismo , Expressão Gênica , Proteínas de Homeodomínio/genética , Proteínas Nucleares , Fatores de Transcrição/genética , Proteínas de Xenopus , Xenopus laevis/embriologia , Sequência de Aminoácidos , Animais , Padronização Corporal , Glândulas Exócrinas/embriologia , Gástrula/metabolismo , Perfilação da Expressão Gênica , Genes Homeobox , Proteínas de Homeodomínio/química , Hibridização In Situ , Dados de Sequência Molecular , Boca/embriologia , Boca/metabolismo , Fatores de Transcrição Box Pareados , Hipófise/embriologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/química , Xenopus laevis/genética , Proteína Homeobox PITX2RESUMO
The mucus secreting cement gland is the anterior-most ectodermal organ of the Xenopus embryo. The homeobox genes Pltx1 and Pitx2c are expressed in the cement gland primordium. Misexpression of both genes induced ectopic cement gland tissue in whole embryos and transcription of the marker genes Xag1 and Xag2 in animal cap explant cultures. Antisense morpholino oligonucleotides against Pitx1 and Pitx2c inhibited ectopic cement gland formation induced by otx2. Gene knock downs generated by morpholino oligonucleotides were specific and could be rescued by coinjection of Pitx mRNAs. These data demonstrate for the first time the requirement of specific genes for cement gland formation by loss-of-function experiments. genesis 30:144--148, 2001.
Assuntos
Coristoma/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/metabolismo , Proteínas Nucleares , Fatores de Transcrição/metabolismo , Xenopus laevis/embriologia , Xenopus laevis/genética , Animais , Coristoma/metabolismo , Ectoderma/metabolismo , Genes Homeobox/genética , Proteínas de Homeodomínio/genética , Proteínas do Tecido Nervoso/metabolismo , Oligonucleotídeos Antissenso/genética , Fatores de Transcrição Otx , Fatores de Transcrição Box Pareados , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transativadores/metabolismo , Fatores de Transcrição/genética , Proteínas de Xenopus/genética , Proteínas de Xenopus/metabolismo , Xenopus laevis/anatomia & histologia , Proteína Homeobox PITX2RESUMO
Seven-transmembrane receptors of the frizzled family can interact with secreted Wnt ligands and transmit Wnt signals into the cell. Dependent on the ligand receptor combination, distinct Wnt pathways are activated. Xenopus frizzled 7 (Xfz7) and Xwnt-8b as well as Human frizzled 5 (Hfz5) and Xwnt-5a can act synergistically in the activation of Wnt/beta-catenin target genes siamois (Xsia) and nodal related 3 (Xnr3) and in the induction of ectopic axes in Xenopus embryos. In order to characterize the role of different protein domains of Xfz7 in Wnt/beta-catenin signaling, chimeric Xfz7/Hfz5 receptors were generated in which the extracellular (N5-TC7) or the intracellular domains (NT7-C5) between Xfz7 and Hfz5 were exchanged. We present evidence that the extracellular domain of Xfz7 can interact with Xwnt-5a and that the intracellular C-terminus can transmit a Wnt/beta-catenin signal. Despite these abilities, Xfz7 and Xwnt-5a do not act synergistically in the activation of Wnt/beta-catenin targets. This implies that the interaction of a frizzled receptor with different ligands can result in distinct cellular responses.
Assuntos
Receptores de Superfície Celular/fisiologia , Receptores Acoplados a Proteínas G , Proteínas de Xenopus , Xenopus/embriologia , Xenopus/fisiologia , Proteínas de Peixe-Zebra , Animais , Sequência de Bases , Primers do DNA/genética , Receptores Frizzled , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Estrutura Terciária de Proteína , Proteínas/genética , Proteínas/fisiologia , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de Superfície Celular/química , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Proteínas Recombinantes de Fusão/metabolismo , Transdução de Sinais , Proteínas Wnt , Proteína Wnt-5a , Xenopus/genéticaRESUMO
Frizzled proteins act as putative Wnt receptors and depending on Wnt/Frizzled interactions distinct intracellular pathways can be activated. The canonical Wnt pathway, triggered by Wnt-1-type ligands, is the best characterized and involves the activation of the cytoplasmic protein Dishevelled (Dsh). The Xenopus frizzled 7 receptor (Xfz7) can act in the canonical (Wnt-1-type) as well as in a non-canonical Wnt pathway which involves the activation of protein kinase C (PKC). In order to analyze the interaction between Xfz7 and Xdsh protein, we tested the effect of overexpression of Xfz7 on the subcellular distribution of Xdsh-myc protein. We demonstrate that Xfz7 can recruit Xdsh-myc to the plasma membrane and target genes of the Wnt-1-type pathway such as siamois and nodal related 3 (Xnr-3) are only activated in the presence of exogenous Xdsh-myc. Xfz7 in combination with Xwnt-8b, however, is able to induce siamois even in the absence of Xdsh-myc. Similar results were obtained after expression of Human frizzled 5 (Hfz5) together with Xwnt-5a but this receptor-ligand combination recruits Xdsh-myc only very poorly to the membrane. These results suggests that the endogenous Xdsh pool and exogenous Xdsh-myc differ in their ability to be recruited by Frizzled receptors.
Assuntos
Fosfoproteínas/metabolismo , Receptores de Superfície Celular/metabolismo , Receptores Acoplados a Proteínas G , Proteínas de Xenopus , Xenopus/embriologia , Proteínas de Peixe-Zebra , Proteínas Adaptadoras de Transdução de Sinal , Animais , Membrana Celular/metabolismo , Proteínas Desgrenhadas , Imunofluorescência , Receptores Frizzled , Proteínas de Homeodomínio/metabolismo , Humanos , Ligantes , Modelos Biológicos , Mutação , Fosfoproteínas/genética , Ligação Proteica , Proteína Quinase C/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , RNA/metabolismo , Receptores de Superfície Celular/genética , Receptores de Neurotransmissores/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Fator de Crescimento Transformador beta/metabolismo , Proteínas Wnt , Proteína Wnt1RESUMO
We have examined the ability of normal and heterotopically transplanted neural crest cells to migrate along cranial neural crest pathways in the axolotl using focal DiI injections and in situ hybridization with the neural crest marker, AP-2. DiI labeling demonstrates that cranial neural crest cells migrate as distinct streams along prescribed pathways to populate the maxillary and mandibular processes of the first branchial arch, the hyoid arch and gill arches 1-4, following migratory pathways similar to those observed in other vertebrates. Another neural crest marker, the transcription factor AP-2, is expressed by premigratory neural crest cells within the neural folds and migrating neural crest cells en route to and within the branchial arches. Rotations of the cranial neural folds suggest that premigratory neural crest cells are not committed to a specific branchial arch fate, but can compensate when displaced short distances from their targets by migrating to a new target arch. In contrast, when cells are displaced far from their original location, they appear unable to respond appropriately to their new milieu such that they fail to migrate or appear to migrate randomly. When trunk neural folds are grafted heterotopically into the head, trunk neural crest cells migrate in a highly disorganized fashion and fail to follow normal cranial neural crest pathways. Importantly, we find incorporation of some trunk cells into branchial arch cartilage despite the random nature of their migration. This is the first demonstration that trunk neural crest cells can form cartilage when transplanted to the head. Our results indicate that, although cranial and trunk neural crest cells have inherent differences in ability to recognize migratory pathways, trunk neural crest can differentiate into cranial cartilage when given proper instructive cues.
Assuntos
Ambystoma mexicanum/embriologia , Encéfalo/embriologia , Crista Neural/fisiologia , Animais , Clonagem Molecular , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Embrião não Mamífero/fisiologia , Morfogênese , Mutação , Crista Neural/citologia , Crista Neural/transplante , Proteínas Recombinantes/metabolismo , Fator de Transcrição AP-2 , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Transplante HeterotópicoRESUMO
Here we report the cloning of a Xenopus frizzled transmembrane receptor, Xfz7, and describe its expression pattern during early embryogenesis. Xfz7 mRNA is provided maternally and zygotic transcription peaks in gastrula stages. At that time, transcripts are preferentially localized to the marginal zone and become restricted to distinct regions of the tadpoles in tailbud stages. Overexpression of Xfz7 in embryos perturbs the morphogenesis of trunk and tail, blocks convergence-extension movements in animal caps induced with activin and dorsal lip explants and decreases cadherin-mediated cell adhesion. Xfz7 can interact specifically with Xwnt-8b and signal in the canonical, dorsalizing Wnt pathway. Overexpression of Xfz7 does not trigger the Wnt-1-type pathway but acts in a non-canonical Wnt or morphogenetic-effector pathway involving the activation of protein kinase C (PKC). Xfz7 seems to be involved in different aspects of Wnt signaling during the course of embryogenesis.
Assuntos
Padronização Corporal/genética , Proteínas Proto-Oncogênicas/metabolismo , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Receptores Acoplados a Proteínas G , Proteínas de Xenopus , Xenopus/genética , Proteínas de Peixe-Zebra , Animais , Adesão Celular/genética , Clonagem Molecular , Ectoderma/metabolismo , Embrião não Mamífero , Feminino , Regulação da Expressão Gênica no Desenvolvimento , Glicoproteínas/genética , Glicoproteínas/metabolismo , Mesoderma/metabolismo , Morfogênese , Proteínas Proto-Oncogênicas/genética , Transdução de Sinais , Cauda/anormalidades , Cauda/embriologia , Proteínas Wnt , Proteína Wnt-5a , Proteína Wnt1 , Proteína Wnt4 , Xenopus/embriologiaRESUMO
During vertebrate left-right development the homeobox gene Pitx2 serves as a mediator between transient nodal signaling in the left lateral plate mesoderm (l-LPM) and asymmetric organ morphogenesis. Misexpression of Pitx2 in chick and frog led to alteration of organ situs. Here we report the presence of different Pitx2 isoforms in mouse and frog. Pitx2c but not Pitx2a or Pitx2b was asymmetrically expressed in the l-LPM, heart and gut, and was specifically induced by nodal in Xenopus animal cap explant cultures and whole embryos. Pitx2c induced its own transcription, suggesting a maintenance mechanism following the down-regulation of nodal in the l-LPM. Pitx2c thus represents the left-specific isoform involved in vertebrate left-right asymmetry.
Assuntos
Padronização Corporal/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas Nucleares , Fatores de Transcrição/genética , Sequência de Aminoácidos , Animais , Embrião não Mamífero/embriologia , Embrião não Mamífero/fisiologia , Desenvolvimento Embrionário e Fetal/genética , Lateralidade Funcional/genética , Camundongos , Dados de Sequência Molecular , Fatores de Transcrição Box Pareados , Isoformas de Proteínas/genética , Alinhamento de Sequência , Xenopus , Proteína Homeobox PITX2RESUMO
Here we report the cloning of zebrafish erm and pea3 cDNAs, which are members of the PEA3 subgroup of Ets transcription factors. The expression patterns of these two genes were examined during zebrafish embryogenesis. Maternal mRNAs of both genes are detectable and transcripts are found ubiquitously until the late blastula, in the marginal zone of gastrula stages and in the presumptive fore- and hindbrain and in the trunk region of early somite stages. Later, erm expression is observed in distinct regions of the forebrain, the mid-/hindbrain boundary, the differentiating rhombomeres, branchial arches, otic vesicles, the pectoral fins, the somites and the tailbud in a dynamic fashion. In contrast, pea3 is not expressed in the rhombomeres but in the Rohon-Beard neurons, the sensory lateral line placodes and the heart.
Assuntos
Encéfalo/embriologia , Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica no Desenvolvimento , Fatores de Transcrição/genética , Peixe-Zebra/embriologia , Animais , Encéfalo/metabolismo , Proteínas de Ligação a DNA/metabolismo , Embrião não Mamífero , Olho/embriologia , Olho/metabolismo , Feminino , Coração/embriologia , Dados de Sequência Molecular , Neurônios/metabolismo , Prosencéfalo/embriologia , Prosencéfalo/metabolismo , Rombencéfalo/embriologia , Rombencéfalo/metabolismo , Fatores de Transcrição/metabolismo , Peixe-Zebra/genética , Proteínas de Peixe-ZebraRESUMO
During vertebrate embryonic development, the organs of the chest and abdomen, heart, lung and gastrointestinal tract, acquire characteristic asymmetric positions with respect to the left-right body axis. In the beginning of the 20th century Hans Spemann and his co-workers described manipulations of amphibian embryos which resulted in inversion of organ laterality in a predictable manner. Hedwig Wilhelmi concluded from these experiments that determinants on the left side of the embryo specify laterality, and Meyer postulated that a mediator should transfer this positional information to the forming heart. In this review we discuss the classical experiments in the light of recent advances in the molecular understanding of left-right development, with a focus on the mediator role of the homeobox gene Pitx2.
Assuntos
Padronização Corporal/fisiologia , Biologia do Desenvolvimento/métodos , Proteínas Nucleares , Transdução de Sinais , Vertebrados/embriologia , Anfíbios/embriologia , Animais , Biologia do Desenvolvimento/tendências , Embrião não Mamífero , Lateralidade Funcional , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas de Homeodomínio/metabolismo , Morfogênese , Proteína Nodal , Fatores de Transcrição Box Pareados , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/metabolismo , Proteína Homeobox PITX2RESUMO
Two secreted signaling molecules, Xwnt-8 and BMP-4, play an essential role in the dorso-ventral patterning of the mesoderm in Xenopus. Here we investigate how the Wnt-8 and the BMP-4 pathways are connected and how they regulate target genes in the lateral and ventral marginal zone. BMP-4 regulates the transcription of Xwnt-8 in a threshold dependent manner. High levels of BMP-4 induce the expression of the Wnt antagonist sizzled in the ventral marginal zone, independent of Xwnt-8 signaling. Xwnt-8 induces the early muscle marker myf-5 in the lateral marginal zone in a BMP independent manner. The expression of the homeobox gene Xvent-1 can be modulated through both the BMP-4 and the Xwnt-8 pathways. The spatial distribution and the level of BMP-4 activity in the lateral and ventral marginal zone is reflected in the dynamic expression pattern of Xwnt-8. The data support the view that Xwnt-8 is involved in the specification of lateral (somitogenic) mesoderm and BMP-4 in the specification of ventral mesoderm.
Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Proteínas de Ligação a DNA , Regulação da Expressão Gênica no Desenvolvimento , Mesoderma/fisiologia , Proteínas Proto-Oncogênicas/metabolismo , Transativadores , Proteínas de Xenopus , Proteínas de Peixe-Zebra , Animais , Padronização Corporal , Proteína Morfogenética Óssea 4 , Gástrula/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Hibridização In Situ , Cloreto de Lítio/farmacologia , Mesoderma/efeitos dos fármacos , Proteínas Musculares/metabolismo , Fator Regulador Miogênico 5 , Proteínas/metabolismo , RNA/metabolismo , Transdução de Sinais , Fatores de Tempo , Transcrição Gênica , Proteínas Wnt , XenopusRESUMO
We report the cloning of a cDNA encoding a Xenopus laevis Ets-type transcription factor. This new Xenopus gene belongs to the PEA3 subfamily of Ets proteins and shows the highest degree of sequence similarity to the mouse and human ER81 genes. The Xenopus ER81 gene (XER81) is transcribed in the embryo after mid blastula transition (MBT) and three transcripts of 3, 4 and 6 kb are detected throughout embryogenesis. XER81 mRNA is localized in the animal pole of the late blastula stage and higher levels of XER81 transcripts are detected in the marginal zone at the onset of gastrulation. In later embryogenesis XER81 transcripts are found in neural crest cells, eyes, otic vesicles and pronephros. The transcription of XER81 can be stimulated by bFGF and eFGF in animal and vegetal cap explants. Expression of the dominant negative FGF receptor mutant in animal caps and embryos blocks XER81 transcription, arguing that the expression of this Ets gene requires active FGF signaling. The spatial overlap of eFGF and XER81 expression domains supports the idea that XER81 transcription could be a marker for regions with active FGF signaling in the embryo.
Assuntos
Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Fatores de Transcrição/fisiologia , Proteínas de Xenopus , Xenopus/embriologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Blastocisto/metabolismo , Northern Blotting , Clonagem Molecular , Proteínas de Ligação a DNA/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Fatores de Transcrição Forkhead , Gástrula/metabolismo , Expressão Gênica , Mesoderma/metabolismo , Modelos Genéticos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Distribuição Tecidual , Ativação TranscricionalRESUMO
Left-right asymmetry in vertebrates is controlled by activities emanating from the left lateral plate. How these signals get transmitted to the forming organs is not known. A candidate mediator in mouse, frog and zebrafish embryos is the homeobox gene Pitx2. It is asymmetrically expressed in the left lateral plate mesoderm, tubular heart and early gut tube. Localized Pitx2 expression continues when these organs undergo asymmetric looping morphogenesis. Ectopic expression of Xnr1 in the right lateral plate induces Pitx2 transcription in Xenopus. Misexpression of Pitx2 affects situs and morphology of organs. These experiments suggest a role for Pitx2 in promoting looping of the linear heart and gut.
Assuntos
Padronização Corporal , Sistema Digestório/embriologia , Genes Homeobox , Coração/embriologia , Proteínas de Homeodomínio/metabolismo , Proteínas Nucleares , Fatores de Transcrição/metabolismo , Vertebrados/embriologia , Anormalidades Múltiplas/embriologia , Ativinas , Sequência de Aminoácidos , Animais , Comunicação Celular , Gástrula , Regulação da Expressão Gênica no Desenvolvimento , Inibinas/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular , Camundongos , Camundongos Mutantes , Dados de Sequência Molecular , Morfogênese , Proteína Nodal , Fatores de Transcrição Box Pareados , Homologia de Sequência de Aminoácidos , Especificidade da Espécie , Distribuição Tecidual , Fator de Crescimento Transformador beta/biossíntese , Fator de Crescimento Transformador beta/metabolismo , Xenopus , Proteínas de Xenopus , Peixe-Zebra , Proteínas de Peixe-Zebra , Proteína Homeobox PITX2RESUMO
Patterning of the marginal zone in the Xenopus embryo has been attributed to interactions between dorsal genes expressed in the organizer and ventral-specific genes. In this antagonistic interplay of activities, BMP-4, a gene that is not expressed in the organizer, provides a strong ventralizing signal. The Xenopus caudal type homeobox gene, Xcad-2, which is expressed around the blastopore with a gap over the dorsal lip, was analyzed as part of the ventral signal. Xcad-2 was shown to efficiently repress during early gastrula stages the dorsal genes gsc, Xnot-2, Otx-2, XFKH1 and Xlim-1, while it positively regulates the ventral genes, Xvent-1 and Xvent-2, with Xpo exhibiting a strong positive response to Xcad-2 overexpression. Xcad-2 was also capable of inducing BMP-4 expression in the organizer region. Support for a ventralizing role for Xcad-2 was obtained from co-injection experiments with the dominant negative BMP receptor which was used to block BMP-4 signaling. Under lack-of-BMP-signaling conditions Xcad-2 could still regulate dorsal and ventral gene expression and restore normal development, suggesting that it can act downstream of BMP-4 signaling or independently of it. Xcad-2 could also inhibit secondary axis formation and dorsalization induced by the dominant negative BMP receptor. Xcad-2 was also shown to efficiently reverse the dorsalizing effects of LiCl. These results place Xcad-2 as part of the ventralizing gene program which acts during early gastrula stages and can execute its ventralizing function in the absence of BMP signaling.
Assuntos
Proteínas Aviárias , Padronização Corporal/genética , Proteínas Morfogenéticas Ósseas/fisiologia , Gástrula/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/fisiologia , Proteínas Repressoras , Fatores de Transcrição , Proteínas de Xenopus/fisiologia , Xenopus laevis/embriologia , Animais , Proteína Morfogenética Óssea 4 , Proteínas de Transporte/biossíntese , Proteínas de Transporte/genética , Proteínas de Ligação a DNA/biossíntese , Proteínas de Ligação a DNA/genética , Fatores de Transcrição Forkhead , Gástrula/fisiologia , Genes Homeobox , Proteína Goosecoid , Proteínas de Homeodomínio/biossíntese , Proteínas de Homeodomínio/genética , Peptídeos e Proteínas de Sinalização Intracelular , Proteínas com Homeodomínio LIM , Microinjeções , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/genética , Organizadores Embrionários , Fatores de Transcrição Otx , Transdução de Sinais , Transativadores/biossíntese , Transativadores/genética , Proteínas de Xenopus/biossíntese , Proteínas de Xenopus/genética , Xenopus laevis/genéticaRESUMO
The Spätzle/Toll signaling pathway controls ventral axis formation in Drosophila by generating a gradient of nuclear Dorsal protein. Dorsal controls the downstream regulators dpp and sog, whose patterning functions are conserved between insects and vertebrates. Although there is no experimental evidence that the upstream events are conserved as well, we set out to ask if a vertebrate embryo can respond to maternal components of the fly Dorsal pathway. Here we demonstrate a dorsalizing activity for the heterologous Easter, Spätzle and Toll proteins in UV-ventralized Xenopus embryos, which is inhibited by a co-injected dominant Cactus variant. We conclude that the Dorsal signaling pathway is a component of the conserved dorsoventral (d/v) patterning system in bilateria.
Assuntos
Padronização Corporal/genética , Proteínas de Drosophila , Proteínas de Insetos/genética , Glicoproteínas de Membrana/genética , Receptores de Superfície Celular , Transdução de Sinais/genética , Xenopus/embriologia , Xenopus/genética , Animais , Vértebra Cervical Áxis/embriologia , Citoplasma/fisiologia , Drosophila/genética , Genes Dominantes/fisiologia , Serina Endopeptidases/genética , Receptores Toll-Like , Xenopus/fisiologiaRESUMO
Expression of the Xenopus Xcad-1 and Xcad-2 genes initiates during early gastrulation exhibiting a dorsoventral asymmetry in their domains of transcription. At mid-gastrulation the ventral preference becomes stronger and the caudal genes take up a posterior localization in their expression, which they will maintain until their downregulation along the dorsal midline. Comparison of the three Xenopus caudal genes revealed a temporal and spatial nested set of expression patterns. The transcription of the caudal genes is sequentially downregulated with the one expressed most caudally (Xcad-2) being shut down first, this sequence is most evident along the dorsal midline. This pattern of expression suggests a role for the caudal genes as posterior determinants along the anteroposterior axis. In chicken, mouse, man and Xenopus three members of the caudal family have been identified in the genome. Even though in Xenopus the Xcad-3 gene has been previously described, in order to obtain a better insight on the role of the caudal genes a comparative study of all three frog genes was performed.
Assuntos
Proteínas Aviárias , Vértebra Cervical Áxis/embriologia , Regulação para Baixo/genética , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Homeodomínio/genética , Proteínas de Xenopus , Xenopus/embriologia , Xenopus/genética , Animais , Padronização Corporal/genética , Proteínas de Drosophila , Proteínas Fetais/biossíntese , Proteínas Fetais/genética , Proteínas de Homeodomínio/biossíntese , Proteínas de Homeodomínio/fisiologia , Fatores de TranscriçãoRESUMO
Patterning events that occur before the mid-blastula transition (MBT) and that organize the spatial pattern of gene expression in the animal hemisphere have been analyzed in Xenopus embryos. We present evidence that genes that play a role in dorsoventral specification display different modes of activation. Using early blastomere explants (16-128-cell stage) cultured until gastrula stages, we demonstrate by RT-PCR analysis that the expression of goosecoid (gsc), wnt-8 and brachyury (bra) is dependent on mesoderm induction. In contrast, nodal-related 3 (nr3) and siamois (sia) are expressed in a manner that is independent of mesoderm induction, however their spatially correct activation does require cortical rotation. The pattern of sia and nr3 expression reveals that the animal half of the 16-cell embryo is already distinctly polarized along the dorsoventral axis as a result of rearrangement of the egg structure during cortical rotation. Similar to the antagonistic activity between the ventral and the dorsal mesoderm, the ventral animal blastomeres can attenuate the expression of nr3 and sia in dorsal animal blastomeres. Our data suggest that no Nieuwkoop center activity at the blastula stage is required for the activation of nr3 and sia in vivo.
Assuntos
Proteínas Fetais , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Repressoras , Proteínas com Domínio T , Xenopus/embriologia , Xenopus/genética , Animais , Sequência de Bases , Blastocisto/citologia , Blastocisto/metabolismo , Blastômeros/citologia , Blastômeros/metabolismo , Padronização Corporal , Proteínas do Citoesqueleto , Primers do DNA/genética , Proteínas de Ligação a DNA/genética , Proteína Goosecoid , Proteínas de Homeodomínio/genética , Mesoderma/citologia , Mesoderma/metabolismo , Reação em Cadeia da Polimerase , Proteínas/genética , Fatores de Transcrição/genética , Proteínas Wnt , Proteínas de Xenopus , Proteínas de Peixe-ZebraRESUMO
The homeobox gene goosecoid has been implicated to play a central role in the Spemann organizer tissue of the vertebrate embryo. Misexpression of goosecoid on the ventral side of a Xenopus laevis gastrula embryo was shown to result in a partial duplication of the primary body axis, reminiscent of the Spemann organizer graft. Normal embryonic development thus requires tight temporal and spatial control of genes instrumental for organizer function. In the present study we investigated the transcriptional control of goosecoid gene expression. Sequence analysis of the mouse and human promoter region revealed the presence of two palindromic binding elements for homeobox genes of the prd type to which goosecoid belongs. We show that Goosecoid protein can bind to these sites in vitro. By using reporter gene constructs of the human and mouse promoter, we demonstrate that Goosecoid can act as a repressor of its own promoter activity in transient co-transfection experiments in mouse P19 cells and in Xenopus embryos. Autorepression depends on the presence of the homeodomain and is mediated through the prd element more proximal to the transcriptional start site. Our results suggest a role for goosecoid in restricting organizer activity in the vertebrate gastrula embryo.
Assuntos
Proteínas de Ligação a DNA/genética , Genes Homeobox , Proteínas de Homeodomínio , Fatores de Transcrição , Sequência de Aminoácidos , Animais , Sequência de Bases , Sítios de Ligação , Linhagem Celular , Clonagem Molecular , DNA Complementar , Proteínas de Ligação a DNA/metabolismo , Embrião não Mamífero/metabolismo , Proteína Goosecoid , Humanos , Camundongos , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Ligação Proteica , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Homologia de Sequência do Ácido Nucleico , Transcrição Gênica , Xenopus laevis/embriologiaRESUMO
Specific signaling molecules play a pivotal role in the induction and specification of tissues during early vertebrate embryogenesis. BMP-4 specifies ventral mesoderm differentiation and inhibits neural induction in Xenopus, whereas three molecules secreted from the organizer, noggin, follistatin and chordin dorsalize mesoderm and promote neural induction. Here we report that follistatin antagonizes the activities of BMP-4 in frog embryos and mouse teratocarcinoma cells. In Xenopus embryos follistatin blocks the ventralizing effect of BMP-4. In mouse P19 cells follistatin promotes neural differentiation. BMP-4 antagonizes the action of follistatin and prevents neural differentiation. In addition we show that the follistatin and BMP-4 proteins can interact directly in vitro. These data provide evidence that follistatin might play a role in modulating BMP-4 activity in vivo.