RESUMO
Background: Congenital hypothyroidism due to thyroid dysgenesis is a frequent congenital endocrine disorder for which the molecular mechanisms remain unresolved in the majority of cases. This situation reflects, in part, our still limited knowledge about the mechanisms involved in the early steps of thyroid specification from the endoderm, in particular the extrinsic signaling cues that regulate foregut endoderm patterning. In this study, we used small molecules and genetic zebrafish models to characterize the role of various signaling pathways in thyroid specification. Methods: We treated zebrafish embryos during different developmental periods with small-molecule compounds known to manipulate the activity of Wnt signaling pathway and observed effects in thyroid, endoderm, and cardiovascular development using whole-mount in situ hybridization and transgenic fluorescent reporter models. We used the antisense morpholino (MO) technique to create a zebrafish acardiac model. For thyroid rescue experiments, bone morphogenetic protein (BMP) pathway induction in zebrafish embryos was obtained by manipulation of heat-shock inducible transgenic lines. Results: Combined analyses of thyroid and cardiovascular development revealed that overactivation of Wnt signaling during early development leads to impaired thyroid specification concurrent with severe defects in the cardiac specification. When using a model of MO-induced blockage of cardiomyocyte differentiation, a similar correlation was observed, suggesting that defective signaling between cardiac mesoderm and endodermal thyroid precursors contributes to thyroid specification impairment. Rescue experiments through transient overactivation of BMP signaling could partially restore thyroid specification in models with defective cardiac development. Conclusion: Collectively, our results indicate that BMP signaling is critically required for thyroid cell specification and identify cardiac mesoderm as a likely source of BMP signals.
Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Proteína Morfogenética Óssea 4/metabolismo , Hipotireoidismo Congênito/metabolismo , Proteínas do Citoesqueleto/metabolismo , Cardiopatias Congênitas/metabolismo , Miócitos Cardíacos/metabolismo , Disgenesia da Tireoide/metabolismo , Glândula Tireoide/metabolismo , Proteínas Wnt/metabolismo , Via de Sinalização Wnt , Proteínas de Peixe-Zebra/metabolismo , Animais , Animais Geneticamente Modificados , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 4/genética , Hipotireoidismo Congênito/genética , Hipotireoidismo Congênito/patologia , Proteínas do Citoesqueleto/genética , Modelos Animais de Doenças , Desenvolvimento Embrionário , Endoderma/anormalidades , Endoderma/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Cardiopatias Congênitas/genética , Cardiopatias Congênitas/patologia , Mesoderma/anormalidades , Mesoderma/metabolismo , Morfolinos/genética , Morfolinos/metabolismo , Miócitos Cardíacos/patologia , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/metabolismo , Disgenesia da Tireoide/genética , Disgenesia da Tireoide/patologia , Glândula Tireoide/anormalidades , Proteínas Wnt/genética , Peixe-Zebra/embriologia , Peixe-Zebra/genética , Peixe-Zebra/metabolismo , Proteínas de Peixe-Zebra/genéticaRESUMO
In the anterior foregut (AFG) of mouse embryos, the transcription factor SOX2 is expressed in the epithelia of the esophagus and proximal branches of respiratory organs comprising the trachea and bronchi, whereas NKX2.1 is expressed only in the epithelia of respiratory organs. Previous studies using hypomorphic Sox2 alleles have indicated that reduced SOX2 expression causes the esophageal epithelium to display some respiratory organ characteristics. In the present study, we produced mouse embryos with AFG-specific SOX2 deficiency. In the absence of SOX2 expression, a single NKX2.1-expressing epithelial tube connected the pharynx and the stomach, and a pair of bronchi developed in the middle of the tube. Expression patterns of NKX2.1 and SOX9 revealed that the anterior and posterior halves of SOX2-deficient AFG epithelial tubes assumed the characteristics of the trachea and bronchus, respectively. In addition, we found that mesenchymal tissues surrounding the SOX2-deficient NKX2.1-expressing epithelial tube changed to those surrounding the trachea and bronchi in the anterior and posterior halves, as indicated by the arrangement of smooth muscle cells and SOX9-expressing cells and by the expression of Wnt4 (esophagus specific), Tbx4 (respiratory organ specific), and Hoxb6 (distal bronchus specific). The impact of mesenchyme-derived signaling on the early stage of AFG epithelial specification has been indicated. Our study demonstrated an opposite trend where epithelial tissue specification causes concordant changes in mesenchymal tissues, indicating a reciprocity of epithelial-mesenchymal interactions.
Assuntos
Esôfago/anormalidades , Trato Gastrointestinal/anormalidades , Organogênese/genética , Fatores de Transcrição SOXB1/deficiência , Traqueia/anormalidades , Animais , Diferenciação Celular/genética , Endoderma/anormalidades , Endoderma/embriologia , Epitélio/embriologia , Esôfago/embriologia , Imunofluorescência , Trato Gastrointestinal/embriologia , Regulação da Expressão Gênica no Desenvolvimento , Mesoderma/embriologia , Camundongos , Camundongos Transgênicos , Traqueia/embriologiaRESUMO
The ZIC transcription factors are key mediators of embryonic development and ZIC3 is the gene most commonly associated with situs defects (heterotaxy) in humans. Half of patient ZIC3 mutations introduce a premature termination codon (PTC). In vivo, PTC-containing transcripts might be targeted for nonsense-mediated decay (NMD). NMD efficiency is known to vary greatly between transcripts, tissues and individuals and it is possible that differences in survival of PTC-containing transcripts partially explain the striking phenotypic variability that characterizes ZIC3-associated congenital defects. For example, the PTC-containing transcripts might encode a C-terminally truncated protein that retains partial function or that dominantly interferes with other ZIC family members. Here we describe the katun (Ka) mouse mutant, which harbours a mutation in the Zic3 gene that results in a PTC. At the time of axis formation there is no discernible decrease in this PTC-containing transcript in vivo, indicating that the mammalian Zic3 transcript is relatively insensitive to NMD, prompting the need to re-examine the molecular function of the truncated proteins predicted from human studies and to determine whether the N-terminal portion of ZIC3 possesses dominant-negative capabilities. A combination of in vitro studies and analysis of the Ka phenotype indicate that it is a null allele of Zic3 and that the N-terminal portion of ZIC3 does not encode a dominant-negative molecule. Heterotaxy in patients with PTC-containing ZIC3 transcripts probably arises due to loss of ZIC3 function alone.
Assuntos
Códon sem Sentido/genética , Síndrome de Heterotaxia/embriologia , Síndrome de Heterotaxia/genética , Proteínas de Homeodomínio/metabolismo , Degradação do RNAm Mediada por Códon sem Sentido/genética , Fatores de Transcrição/metabolismo , Alelos , Animais , Sequência de Bases , Núcleo Celular/metabolismo , Difusão , Embrião de Mamíferos/anormalidades , Embrião de Mamíferos/metabolismo , Embrião de Mamíferos/patologia , Endoderma/anormalidades , Endoderma/embriologia , Gastrulação/genética , Proteínas de Homeodomínio/genética , Humanos , Mesoderma/anormalidades , Mesoderma/embriologia , Camundongos , Camundongos Mutantes , Dados de Sequência Molecular , Proteínas Mutantes/metabolismo , Mutação/genética , Organogênese/genética , Estabilidade Proteica , Sítios de Splice de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/genética , Transcrição Gênica , beta Catenina/metabolismoRESUMO
Two cases of endodermal cyst of the posterior fossa are reported. A 12-year-old girl presented with severe headache and vomiting caused by increased intracranial pressure. Computed tomography and magnetic resonance (MR) imaging showed a cystic mass occupying the ambient and quadrigeminal cisterns. A 65-year-old woman presented with dizziness, and MR imaging revealed a cystic mass in the posterior fossa. The two patients underwent surgery for decompression and resection of the cyst. Surgical specimens of the cyst walls consisted of a single layer of ciliated columnar epithelium. The diagnoses were endodermal cyst. The optimal surgical goal is total resection of the cyst wall, but the cyst wall sometimes tightly adheres to the adjacent nerves, vessels, and vital structures. The cyst must communicate adequately with the surrounding cerebrospinal fluid space, and a newly closed cyst space must be avoided, by the widest possible resection of the cyst wall.
Assuntos
Fossa Craniana Posterior/cirurgia , Endoderma/patologia , Defeitos do Tubo Neural/cirurgia , Procedimentos Neurocirúrgicos/métodos , Idoso , Criança , Fossa Craniana Posterior/diagnóstico por imagem , Fossa Craniana Posterior/patologia , Descompressão Cirúrgica/métodos , Endoderma/anormalidades , Feminino , Humanos , Hipertensão Intracraniana/etiologia , Defeitos do Tubo Neural/diagnóstico por imagem , Defeitos do Tubo Neural/patologia , RadiografiaRESUMO
The mammalian respiratory system, consisting of both trachea and lung, initiates from the foregut endoderm. The molecular program that instructs endodermal cells to adopt the respiratory fate is not fully understood. Here we show that conditional inactivation of beta-Catenin (also termed Ctnnb1) in foregut endoderm leads to absence of both the trachea and lung due to a failure in maintaining the respiratory fate. In converse, conditional expression of an activated form of beta-Catenin leads to expansion of Nkx2.1, an early marker for the trachea and lung, into adjacent endoderm including the stomach epithelium. Analyses of these mutants show that the loss or gain of trachea/lung progenitor identity is accompanied by an expansion or contraction of esophagus/stomach progenitor identity, respectively. Our findings reveal an early role for beta-Catenin in the establishment of respiratory progenitors in mouse foregut endoderm.
Assuntos
Pulmão/anormalidades , Sistema Respiratório/embriologia , Traqueia/anormalidades , beta Catenina/fisiologia , Animais , Embrião de Mamíferos/anormalidades , Embrião de Mamíferos/metabolismo , Endoderma/anormalidades , Endoderma/metabolismo , Feminino , Fator 10 de Crescimento de Fibroblastos/genética , Fator 10 de Crescimento de Fibroblastos/metabolismo , Imunofluorescência , Regulação da Expressão Gênica no Desenvolvimento , Hibridização In Situ , Pulmão/metabolismo , Masculino , Camundongos , Camundongos Knockout , Sistema Respiratório/metabolismo , Anormalidades do Sistema Respiratório/genética , Anormalidades do Sistema Respiratório/fisiopatologia , Transdução de Sinais , Traqueia/metabolismo , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , Proteínas Wnt/fisiologia , beta Catenina/genética , beta Catenina/metabolismoRESUMO
We report the isolation of a recessive ENU-induced short-tailed mutant in the ascidian Ciona intestinalis that is the product of a premature stop in the brachyury gene. Notochord differentiation and morphogenesis are severely disrupted in the mutant line. At the larval stage, variable degrees of ectopic endoderm staining were observed in the homozygous mutants, indicating that loss of brachyury results in stochastic fate transformation. In post-metamorphosis mutants, a uniform defect in tail resorption was observed, together with variable defects in digestive tract development. Some cells misdirected from the notochord lineage were found to be incorporated into definitive endodermal structures, such as stomach and intestine.
Assuntos
Ciona intestinalis/crescimento & desenvolvimento , Endoderma/anormalidades , Endoderma/crescimento & desenvolvimento , Proteínas Fetais/deficiência , Mutação/genética , Proteínas com Domínio T/deficiência , Sequência de Aminoácidos , Animais , Sequência de Bases , Biomarcadores/metabolismo , Linhagem da Célula , Ciona intestinalis/citologia , Análise Mutacional de DNA , Etilnitrosoureia , Proteínas Fetais/química , Proteínas Fetais/genética , Trato Gastrointestinal/anormalidades , Trato Gastrointestinal/crescimento & desenvolvimento , Metamorfose Biológica , Dados de Sequência Molecular , Notocorda/citologia , Especificidade de Órgãos , Proteínas com Domínio T/química , Proteínas com Domínio T/genética , Cauda/anormalidades , Cauda/crescimento & desenvolvimentoRESUMO
In the nematode, C. elegans, the bZIP/homeodomain transcription factor SKN-1 and the Wnt effector TCF/POP-1 are central to the maternal specification of the endomesoderm prior to gastrulation. The 8-cell stage blastomere MS is primarily a mesodermal precursor, giving rise to cells of the pharynx and body muscle among others, while its sister E clonally generates the entire endoderm (gut). In C. elegans, loss of SKN-1 results in the absence of MS-derived tissues all of the time, and loss of gut most of the time, while loss of POP-1 results in a mis-specification of MS as an E-like cell, resulting in ectopic gut. We show that in C. briggsae, RNAi of skn-1 results in a stronger E defect but no apparent MS defect, while RNAi of pop-1 results in loss of gut and an apparent E to MS transformation, the opposite of the pop-1 knockdown phenotype seen in C. elegans. The difference in pop-1(-) phenotypes correlates with changes in how the endogenous endoderm-specifying end genes are regulated by POP-1 in the two species. Our results suggest that integration of Wnt-dependent and Wnt-independent cell fate specification pathways within the Caenorhabditis genus can occur in different ways.
Assuntos
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/embriologia , Caenorhabditis/embriologia , Proteínas de Ligação a DNA/metabolismo , Endoderma/embriologia , Proteínas de Grupo de Alta Mobilidade/metabolismo , Mesoderma/embriologia , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Animais , Animais Geneticamente Modificados , Padronização Corporal , Caenorhabditis/genética , Caenorhabditis elegans/metabolismo , Embrião não Mamífero/anormalidades , Embrião não Mamífero/metabolismo , Endoderma/anormalidades , Regulação da Expressão Gênica no Desenvolvimento , Mesoderma/metabolismo , Modelos Biológicos , Dados de Sequência Molecular , Faringe/anormalidades , Fenótipo , Interferência de RNA , Homologia de Sequência de Aminoácidos , Proteínas Wnt/metabolismoRESUMO
Four members of the TEAD/TEF family of transcription factors are expressed widely in mouse embryos and adult tissues. Although in vitro studies have suggested various roles for TEAD proteins, their in vivo functions remain poorly understood. Here we examined the role of Tead genes by generating mouse mutants for Tead1 and Tead2. Tead2(-/-) mice appeared normal, but Tead1(-/-); Tead2(-/-) embryos died at embryonic day 9.5 (E9.5) with severe growth defects and morphological abnormalities. At E8.5, Tead1(-/-); Tead2(-/-) embryos were already small and lacked characteristic structures such as a closed neural tube, a notochord, and somites. Despite these overt abnormalities, differentiation and patterning of the neural plate and endoderm were relatively normal. In contrast, the paraxial mesoderm and lateral plate mesoderm were displaced laterally, and a differentiated notochord was not maintained. These abnormalities and defects in yolk sac vasculature organization resemble those of mutants for Yap, which encodes a coactivator of TEAD proteins. Moreover, we demonstrated genetic interactions between Tead1 and Tead2 and Yap. Finally, Tead1(-/-); Tead2(-/-) embryos showed reduced cell proliferation and increased apoptosis. These results suggest that Tead1 and Tead2 are functionally redundant, use YAP as a major coactivator, and support notochord maintenance as well as cell proliferation and survival in mouse development.