RESUMEN
Mastocytosis is a heterogeneous disease characterized by an abnormal accumulation of mast cells (MCs) in 1 or several organs. Although a somatic KIT D816V mutation is detected in â¼85% of patients, attempts to demonstrate its oncogenic effect alone have repeatedly failed, suggesting that additional pathways are involved in MC transformation. From 3 children presenting with both Greig cephalopolysyndactyly syndrome (GCPS, Mendelian Inheritance in Man [175700]) and congenital mastocytosis, we demonstrated the involvement of the hedgehog (Hh) pathway in mastocytosis. GCPS is an extremely rare syndrome resulting from haploinsufficiency of GLI3, the major repressor of Hh family members. From these familial cases of mastocytosis, we demonstrate that the Hh pathway is barely active in normal primary MCs and is overactive in neoplastic MCs. GLI3 and KIT mutations had a synergistic, tumorigenic effect on the onset of mastocytosis in a GCPS mouse model. Finally, Hh inhibitors suppressed neoplastic MC proliferation in vitro and extend the survival time of mice with aggressive systemic mastocytosis (ASM). This work revealed, for the first time, the involvement of Hh signaling in the pathophysiology of mastocytosis and demonstrated the cooperative effects of the KIT and Hh oncogenic pathways in mice with ASM, leading to the identification of new promising therapeutic targets.
Asunto(s)
Acrocefalosindactilia/complicaciones , Proteínas Hedgehog/metabolismo , Mastocitosis/complicaciones , Transducción de Señal , Acrocefalosindactilia/metabolismo , Animales , Células Cultivadas , Niño , Humanos , Mastocitosis/metabolismo , Ratones Endogámicos C57BL , Ratones SCID , Células Tumorales CultivadasRESUMEN
Inducible serum proteins whose concentrations oscillate between nontolerogenic and tolerogenic levels pose a particular challenge to the maintenance of self-tolerance. Temporal restrictions of intrathymic antigen supply should prevent continuous central tolerization of T cells, in analogy to the spatial limitation imposed by tissue-restricted antigen expression. Major acute-phase proteins such as human C-reactive protein (hCRP) are typical examples for such inducible self-antigens. The circulating concentration of hCRP, which is secreted by hepatocytes, is induced up to 1,000-fold during an acute-phase reaction. We have analyzed tolerance to hCRP expressed in transgenic mice under its autologous regulatory regions. Physiological regulation of basal levels (<10(-9) M) and inducibility (>500-fold) are preserved in female transgenics, whereas male transgenics constitutively display induced levels. Surprisingly, crossing of hCRP transgenic mice to two lines of T cell receptor transgenic mice (specific for either a dominant or a subdominant epitope) showed that tolerance is mediated by intrathymic deletion of immature thymocytes, irrespective of widely differing serum levels. In the absence of induction, hCRP expressed by thymic medullary epithelial cells rather than liver-derived hCRP is necessary and sufficient to induce tolerance. Importantly, medullary epithelial cells also express two homologous mouse acute-phase proteins. These results support a physiological role of "ectopic" thymic expression in tolerance induction to acute-phase proteins and possibly other inducible self-antigens and have implications for delineating the relative contributions of central versus peripheral tolerance.
Asunto(s)
Proteína C-Reactiva/inmunología , Linfocitos T CD4-Positivos/inmunología , Autotolerancia/inmunología , Timo/inmunología , Proteínas de Fase Aguda/inmunología , Reacción de Fase Aguda/inmunología , Animales , Células Clonales/inmunología , Cruzamientos Genéticos , Epítopos/inmunología , Femenino , Citometría de Flujo , Regulación de la Expresión Génica/genética , Humanos , Hígado/metabolismo , Masculino , Ratones , Ratones Transgénicos , Fragmentos de Péptidos/inmunologíaRESUMEN
The members of the multigene family of intermediate filament (IF) proteins are expressed in various combinations and amounts that are specific for a given pathway or state of differentiation. Previous experiments in which the cell type-specific IF cytoskeleton was altered by introducing foreign IF proteins into cultured cells or certain tissues of transgenic animals have shown a remarkable tolerance, without detectable interference with cell functions. To examine the importance of the cell type-specific cytokeratin (CK) IF pattern, we have studied the ectopic expression of CK genes in different epithelia of transgenic mice. Here we report changes observed in the beta cells of pancreatic islets expressing the genes for human epidermal CKs 1 and/or 10 brought under control of the rat insulin promoter. Both genes were efficiently expressed, resulting in the appearance of numerous and massive bundles of aggregated IFs, resembling those of epidermal keratinocytes. While the synthesis of epidermal CK 10 was readily accommodated and compatible with cell function, mice expressing CK 1 in their beta cells, alone or in combination with CK 10, developed a special form of diabetes characterized by a drastic reduction of insulin-secretory vesicles and of insulin-and CK 1-producing cells. In many CK 1-producing cells, accumulations of fibrous or granular material containing CK 1 were also seen in the nucleus. This demonstration of functional importance of the specific CK-complement in an epithelial cell indicates a contribution of cell type-specific factors to cytoplasmic IF compartmentalization and that the specific CK complement can be crucial for functions and longevity of a given kind of epithelium.
Asunto(s)
Citoesqueleto/ultraestructura , Insulina/biosíntesis , Islotes Pancreáticos/metabolismo , Queratinas/biosíntesis , Piel/metabolismo , Células 3T3 , Animales , Glucemia/metabolismo , Núcleo Celular/metabolismo , Núcleo Celular/ultraestructura , Células HeLa , Humanos , Insulina/análisis , Islotes Pancreáticos/ultraestructura , Queratinas/análisis , Queratinas/genética , Ratones , Ratones Transgénicos , Microscopía Electrónica , Microscopía Inmunoelectrónica , Plásmidos , Mapeo Restrictivo , TransfecciónRESUMEN
A novel mouse homeobox-containing gene, Nkx-2.2, has been isolated. Nkx-2.2 is a member of a family of genes whose homeodomains are homologous to that of the Drosophila NK-2 gene. Nkx-2.2 transcripts are found in localized domains of the brain during mouse embryogenesis. Nkx-2.2 expression in the brain abuts and partially overlaps with the expression domains of two other related homeobox-containing genes, TTF-1 and Dlx. The expression domains of the three genes in the developing prosencephalon coincide with anatomical boundaries, particularly apparent in the diencephalon. This result raises the possibility that these genes may specify regional differentiation of the developing diencephalon into its anatomically and functionally defined subregions. Nkx-2.2 may be involved in specifying diencephalic neuromeric boundaries.
Asunto(s)
Regulación de la Expresión Génica/genética , Genes Homeobox/genética , Prosencéfalo/embriología , Animales , Secuencia de Bases , Northern Blotting , Southern Blotting , Mapeo Cromosómico , ADN/genética , Diencéfalo/embriología , Diencéfalo/fisiología , Regulación de la Expresión Génica/fisiología , Genes Homeobox/fisiología , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Morfogénesis/fisiología , Hibridación de Ácido Nucleico , Prosencéfalo/fisiología , Transcripción Genética/genéticaRESUMEN
Addition of serum to mitogen-starved cells activates the cellular immediate-early gene (IEG) response. Serum response factor (SRF) contributes to such mitogen-stimulated transcriptional induction of many IEGs during the G0-G1 cell cycle transition. SRF is also believed to be essential for cell cycle progression, as impairment of SRF activity by specific antisera or antisense RNA has previously been shown to block mammalian cell proliferation. In contrast, Srf(-/-) mouse embryos grow and develop up to E6.0. Using the embryonic stem (ES) cell system, we demonstrate here that wild-type ES cells do not undergo complete cell cycle arrest upon serum withdrawal but that they can mount an efficient IEG response. This IEG response, however, is severely impaired in Srf(-/-) ES cells, providing the first genetic proof that IEG activation is dependent upon SRF. Also, Srf(-/-) ES cells display altered cellular morphology, reduced cortical actin expression, and an impaired plating efficiency on gelatin. Yet, despite these defects, the proliferation rates of Srf(-/-) ES cells are not substantially altered, demonstrating that SRF function is not required for ES cell cycle progression.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Genes Inmediatos-Precoces , Proteínas Inmediatas-Precoces , Proteínas Nucleares/metabolismo , Animales , Secuencia de Bases , Ciclo Celular , Ensayo de Unidades Formadoras de Colonias , Cartilla de ADN/genética , Proteínas de Unión al ADN/genética , Proteína 1 de la Respuesta de Crecimiento Precoz , Embrión de Mamíferos/citología , Embrión de Mamíferos/metabolismo , Desarrollo Embrionario y Fetal/genética , Genes fos , Ratones , Ratones Noqueados , Microscopía Electrónica de Rastreo , Proteínas Nucleares/genética , Factor de Respuesta Sérica , Transducción de Señal , Células Madre/citología , Células Madre/metabolismo , Factores de Transcripción/genéticaRESUMEN
Transgenic mice have been generated with an inducible SV40 t/T antigen construct with the aim of analysing the early changes that take place in the course of liver tumorigenesis. The strictly liver-specific human C-reactive protein (CRP) gene promoter was chosen for the control of the transgene expression because this promoter can be turned on transiently by injection of bacterial lipopolysaccharide. Among 10 independently derived CRP-Tag mouse lines five showed inducible expression of the CRP-Tag transgene in liver. However, only one had a tight control of the transgene with virtually no expression under physiological conditions and high levels of Tag expression after stimulation. Females of this line were used to analyse the progression of liver alterations upon repeated induction of the t/T antigen for different lengths of time. The first signs of transgene-induced liver alterations could be monitored by the activation of the marker enzyme gamma-glutamyltranspeptidase 30 days after the start of the induction program. After 90 days hepatocellular carcinomas were already detectable. Thus, CRP-Tag mice constitute an excellent system to analyse the sequential events that take place during liver carcinogenesis.
Asunto(s)
Antígenos Transformadores de Poliomavirus/genética , Proteína C-Reactiva/genética , Neoplasias Hepáticas Experimentales/etiología , Animales , Femenino , Expresión Génica , Lipopolisacáridos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Regiones Promotoras Genéticas , ARN Mensajero/análisis , Factores Sexuales , Factores de TiempoRESUMEN
The proto-oncogene c-fos has been isolated as the cellular homolog of the v-fos gene found in the osteosarcoma inducing FBR- and FBJ-murine sarcoma viruses (MSV). Expression of the c-fos gene in transgenic mice leads to the development of bone lesions of which about half progress to bone tumors mainly chondrosarcomas. The tumors display a strong preference for males and have a latency with a mean of 9.5 months. However, also mice without visible lesions develop bone tumors with the same sex preference and latency. These consequences of c-fos expression are independent of the chosen promoter but dependent on a replacement of 3' noncoding sequences of c-fos by a long terminal repeat (LTR) of the FBJ-MSV virus.
Asunto(s)
Neoplasias Óseas/etiología , Proteínas Proto-Oncogénicas/genética , Proto-Oncogenes , Animales , Neoplasias Óseas/patología , Femenino , Masculino , Ratones , Ratones Transgénicos , Proteínas Proto-Oncogénicas c-fos , Secuencias Repetitivas de Ácidos Nucleicos , Virus del Sarcoma Murino/genética , Factores SexualesRESUMEN
Herpesvirus saimiri is an oncogenic virus causing rapid T-cell lymphomas in New World primates and rabbits. Deletion analysis of one strain of H saimiri has indicated an open reading frame, StpA, necessary for oncongenicity in monkeys. We have investigated the function of StpA in tumor induction by the generation of transgenic mice. Expression of two different constructs caused the development of peripheral lymphomas. The infiltrating cells were of T-cell origin, expressing mainly the CD4 phenotype and restricted sets of V beta chains. Thus, StpA is not only necessary for the oncogenicity of Herpesvirus saimiri, but is also sufficient for the induction of peripheral pleomorphic T-cell lymphomas.
Asunto(s)
Proteínas Bacterianas/genética , Proteínas de Unión al ADN , Proteínas de Escherichia coli , Herpesvirus Saimiriino 2/genética , Linfoma de Células T/genética , Linfoma de Células T/virología , Ratones Transgénicos/genética , Ratones Transgénicos/virología , Chaperonas Moleculares , Proteínas Oncogénicas Virales/genética , Animales , Secuencia de Bases , Northern Blotting , Complejo CD3 , Antígenos CD4 , Clonación Molecular , Expresión Génica , Inmunohistoquímica , Activación de Linfocitos , Ratones , Datos de Secuencia Molecular , Fenotipo , Regiones Promotoras Genéticas , Receptores de Antígenos de Linfocitos T/química , Receptores de Antígenos de Linfocitos T/genéticaRESUMEN
The potential of tissue inhibitors of metalloproteinases (TIMPs) to inhibit neoplastic progression has been postulated from studies of genetically manipulated cells. To investigate whether the TIMP-1 expressed in a host tissue suppresses cancer in vivo and to identify the affected stages, we developed transgenic mice with constitutive overexpression or reduction of TIMP-1 in the liver. In double transgenic experiments, the TIMP-1 lines were crossed with a second transgenic line which expresses the Simian Virus 40t/T antigen (TAg). This viral oncogene leads to heritable development of hepatocellular carcinomas with a 100% incidence. Effects of TIMP-1 coexpression on the TAg-induced neoplasms were determined at the tissue and cellular level. Here, we report that overexpression of hepatic TIMP-1 blocked the development of TAg-induced hepatocellular carcinomas. High TIMP-1 levels inhibited not only the later stages in tumor development (growth and angiogenesis), but also events associated with tumor initiation (altered hepatocyte cytology and tissue architecture). We further show that an antisense-mediated reduction of TIMP-1 resulted in a more rapid tumor initiation and progression. These data demonstrate that intrinsic TIMP-1 levels contribute to a tissue's susceptibility to viral oncogene-induced tumorigenesis.
Asunto(s)
Antígenos Transformadores de Poliomavirus/fisiología , Glicoproteínas/fisiología , Neoplasias Hepáticas Experimentales/prevención & control , Animales , Antígenos Transformadores de Poliomavirus/biosíntesis , Antígenos Transformadores de Poliomavirus/genética , Transformación Celular Neoplásica , ADN Complementario/genética , Femenino , Glicoproteínas/genética , Glicoproteínas/metabolismo , Hígado/metabolismo , Hígado/patología , Neoplasias Hepáticas Experimentales/inducido químicamente , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Inhibidores Tisulares de MetaloproteinasasRESUMEN
Within the tumor-stromal microenvironment a disrupted balance between matrix metalloproteinases (MMPs) and their inhibitors compromises the integrity of the extracellular matrix and promotes malignancy. Tissue inhibitors of metalloproteinases (TIMPs) have been linked to tumor suppression in studies of genetically altered tissue culture cells and in analyses of clinical specimens in situ. We generated transgenic mice as a model system to test the relationship between TIMP-1 levels in a host organ and susceptibility to experimentally targeted metastasis. Ectopically overexpressed TIMP-1 in the brain resulted in a tissue microenvironment with elevated protein levels of this natural MMP inhibitor. Metastatic challenge provided by lacZ-tagged fibrosarcoma cells permitted high-resolution analysis of metastatic load and pattern. We found that elevated host TIMP-1 imposed resistance to experimental metastasis of fibrosarcoma: In TIMP-1 overexpressing mice, brain metastases were significantly reduced by 75% compared to wild-type littermates. Our findings demonstrate that ectopic TIMP-1 expression efficiently exerts a suppressive effect on metastasizing tumor cells.
Asunto(s)
Neoplasias Encefálicas/secundario , Fibrosarcoma/secundario , Metástasis de la Neoplasia/genética , Inhibidor Tisular de Metaloproteinasa-1/biosíntesis , Animales , Inmunidad Innata , Ratones , Ratones Transgénicos , Inhibidor Tisular de Metaloproteinasa-1/genéticaRESUMEN
To investigate the role of herpesviral genes in tumourigenesis, transgenic mice were generated expressing STP-C, a transformation associated protein of the lymphoma inducing herpesvirus saimiri. Epithelial tumours developed in the salivary gland, pancreas, thymus and liver of transgenic mice within the first weeks of life. Thus, the target cells for tumour formation in the transgenic mice were surprisingly different from those of the herpesvirus from which the oncogene was derived. Our results identify STP-C as a herpesvirus oncogene sufficient for tumour induction without the cooperation of other viral gene products. Furthermore, the results demonstrate pleiotropic transforming capabilities of the STP-C oncogene and suggest that the specificity of lymphoma induction by the virus is determined by factors other than the oncogene itself.
Asunto(s)
Transformación Celular Neoplásica , Herpesvirus Saimiriino 2/genética , Neoplasias/etiología , Proteínas Oncogénicas Virales/genética , Oncogenes , Animales , Secuencia de Bases , Epitelio/patología , Activación de Linfocitos , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , FenotipoRESUMEN
We have isolated and analysed embryonic stem (ES) cell clones after electroporation with a gene trap vector. Clones were screened for changes in their lacZ reporter gene activity upon in vitro differentiation. The cDNA of one of the trapped transcripts, T10-2A2, was isolated and analysed in detail. Although not expressed constitutively in differentiating ES cells, the transcript was present in most organs of adult mice and widely expressed in midgestation mouse embryos. Zoo blot analysis indicated a conservation of this novel gene in yeast, rat and human.
Asunto(s)
ARN Mensajero/química , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Evolución Biológica , Diferenciación Celular , Clonación Molecular , Secuencia Conservada , Embrión de Mamíferos , Humanos , Operón Lac , Ratones , Datos de Secuencia Molecular , Ratas , Células Madre/metabolismoRESUMEN
The zinc finger gene Gli3 plays a role in limb and brain development. To facilitate the molecular analysis of different mouse mutations of this gene, the murine cDNA was isolated and sequenced. This 5113 bp cDNA encodes a putative protein of 1596 amino acids. Comparison of the murine and human GLI3 cDNA revealed an overall homology of 85% between the deduced amino acid sequences. More importantly, several regions of the protein, including the zinc fingers, are more highly conserved ( > 95%), suggesting that these represent functional domains in the Gli3 protein.
Asunto(s)
ADN Complementario/química , Proteínas de Unión al ADN/genética , Proteínas del Tejido Nervioso , Proteínas Represoras , Factores de Transcripción/genética , Proteínas de Xenopus , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Proteínas de Unión al ADN/química , Humanos , Factores de Transcripción de Tipo Kruppel , Ratones , Datos de Secuencia Molecular , Proteína Gli3 con Dedos de ZincRESUMEN
The most obvious phenotype of Ft/+ mice is a syndactyly of fore limbs characterised by a fusion of the tips of digits 1 to 4. The tempospatial expression of genes involved in limb development revealed that patterning of Ft/+ limb buds is not affected by the mutation. However, an upregulation of Bmp4 in the anterior-distal region of the limb bud at d12.0 of embryonic development is accompanied by a loss of Fgf8 expression in the distal part of the AER. Downstream target genes of Bmp action such as Msx1 and 2 are upregulated. This induction of the signalling cascade indicates ectopic expression of functional Bmp4. Nevertheless, analysis of physical parameters of bones from adult mice revealed a reduction of the bone mass of the autopod. The data suggest a negative effect of Bmp4 on Fgf8 expression and a positive influence on the induction of bone elements.
Asunto(s)
Proteínas Morfogenéticas Óseas/genética , Factores de Crecimiento de Fibroblastos/genética , Sindactilia/genética , Factor de Crecimiento Transformador beta , Animales , Tipificación del Cuerpo/genética , Proteína Morfogenética Ósea 2 , Proteína Morfogenética Ósea 4 , Proteína Morfogenética Ósea 7 , Proteínas Morfogenéticas Óseas/metabolismo , Proteínas Portadoras , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Ectodermo/metabolismo , Extremidades/anatomía & histología , Extremidades/embriología , Factor 8 de Crecimiento de Fibroblastos , Factores de Crecimiento de Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Proteínas de Homeodominio/metabolismo , Factores de Transcripción de Tipo Kruppel , Esbozos de los Miembros/metabolismo , Factor de Transcripción MSX1 , Ratones , Ratones Endogámicos , Ratones Mutantes , Ratones Transgénicos , Mutación , Proteínas Oncogénicas/genética , Tamaño de los Órganos/genética , Proteínas/genética , Proteínas/metabolismo , Transactivadores , Factores de Transcripción/genética , Proteína con Dedos de Zinc GLI1 , Proteína Gli2 con Dedos de ZincRESUMEN
We have investigated the expression of left/right (L/R) asymmetry markers, nodal and lefty, in the situs inversus mouse mutant Fused toes (Ft). Both genes exhibited bilateral expression in the lateral plate mesoderm (LPM) at developmental stages whereas in wildtype embryos these genes were found to be expressed exclusively in the left LPM. Inspection of tail location and primitive heart tube looping, structures known to be handed in their orientation, documented a random orientation of these structures. Crossing of the Ft mutation into a different genetic background resulted in a strong reduction of this random orientation. Although the major fraction of these individuals still displayed nodal and lefty on both sides of the LPM, expression was almost always found to be weaker in the right LPM. These results suggest that the establishment of asymmetry is independent of nodal or lefty signals. However, handed asymmetry, which means consistent L/R differences, such as the dextral looping of the primitive heart tube or the right-oriented tail, is directed by differences in the L/R expression pattern of these two genes.
Asunto(s)
Desarrollo Embrionario y Fetal/genética , Lateralidad Funcional/genética , Regulación del Desarrollo de la Expresión Génica , Mutación , Animales , Corazón/embriología , Homocigoto , Factores de Determinación Derecha-Izquierda , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Proteína Nodal , Fenotipo , Proteínas/genética , Cola (estructura animal)/embriología , Factor de Crecimiento Transformador beta/genéticaRESUMEN
dkk-1 has recently been identified as a secreted protein in Xenopus laevis which is sufficient and necessary to cause head induction by antagonizing Wnt signalling (Glinka et al., 1998, Nature 391, 357-362). Consistent with such a role dkk-1 is expressed in the Spemann organizer of the early frog gastrula. Later, expression can be observed in an endomesodermal domain corresponding to the prospective prechordal plate, in two longitudinal stripes flanking the anterior chordamesoderm and in the precursors of the liver. At late neurula stage expression occurs in the prechordal plate adjacent to the prospective forebrain and eyes and in a stripe corresponding to the forming somites. dkk-1 is part of a gene family with at least three family members which is conserved between species. Its mouse homologue, Dkk-1, is first expressed at embryonic day (E) 6.5 in mesodermal cells adjacent to the embryonic/extraembryonic junction. Starting at E7.5 transcripts can be detected in the head mesoderm and at E8.5 additionally in developing somites (Glinka et al., 1998, Nature 391, 357-362). In this study we focus on the highly dynamic pattern of Dkk-1 mRNA distribution during mouse limb development from E9.0-E14.5. The other currently known family members, Dkk-2 and -3, are not expressed in the limb bud before E11.5 (C. Niehrs, pers. commun.) while the limb pattern is established. We show that Dkk-1 expression starts with the first sign of forelimb budding, whereas in the presumptive hindlimb region transcription becomes already apparent before the limb starts to bud out. Expression then becomes confined to two mesenchymal domains at E10.5 and E11.5. Using double-whole mount in situ hybridization we show that the posterior Dkk-1 expression domain initially overlaps with that of Shh, one of the key signalling molecules in limb development. Later, the two expression domains become separated. At E12.5-E14.5 Dkk-1 transcripts are restricted to the interdigital mesenchyme.
Asunto(s)
Extremidades/embriología , Proteínas/genética , Animales , Regulación del Desarrollo de la Expresión Génica , Péptidos y Proteínas de Señalización Intercelular , Esbozos de los Miembros , Mesodermo/fisiología , Ratones , Ratones EndogámicosRESUMEN
Sonic hedgehog (Shh) expression in the developing limb is associated with the zone of polarising activity (ZPA), and both are restricted to the posterior part of the limb bud. We show that the expression patterns of Shh and Gli3, a member of the Gli-family believed to function in transcriptional control, appear to be mutually exclusive in limb buds of mouse embryos. In the polydactyly mouse mutant extra toes (Xt), possessing a null mutation of Gli3, Shh is additionally expressed in the anterior region of the limb bud. The transcript of Ptc, the putative receptor for Shh protein, can be detected anteriorly as well. Other genes known to be involved in limb outgrowth and patterning, like Fibroblast growth factor (Fgf), Bone morphogenetic protein (Bmp), and Hoxd are misexpressed in relation to the ectopic Shh expression domain in Xt limb buds. This data suggest that Gli3 is a regulator of Shh expression in mouse limb development.
Asunto(s)
Proteínas Morfogenéticas Óseas/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteínas de Drosophila , Extremidades/fisiología , Factores de Crecimiento de Fibroblastos/metabolismo , Regulación del Desarrollo de la Expresión Génica , Proteínas del Tejido Nervioso , Proteínas/metabolismo , Proteínas Represoras , Transactivadores , Factores de Transcripción , Factor de Crecimiento Transformador beta , Proteínas de Xenopus , Animales , Proteína Morfogenética Ósea 2 , Proteína Morfogenética Ósea 4 , Proteína Morfogenética Ósea 7 , Extremidades/embriología , Factor 4 de Crecimiento de Fibroblastos , Factor 8 de Crecimiento de Fibroblastos , Sustancias de Crecimiento/metabolismo , Proteínas Hedgehog , Proteínas de Homeodominio/metabolismo , Hibridación in Situ , Proteínas de Insectos/metabolismo , Factores de Transcripción de Tipo Kruppel , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C3H , Mutación , Polidactilia/genética , Proteínas Proto-Oncogénicas/metabolismo , Receptores de Superficie Celular , Proteína Gli3 con Dedos de ZincRESUMEN
Signaling of TGF-beta superfamily members is tightly controlled by an elaborate network of regulators (for recent review see Trends Genet. 15 (1999) 3; Genes Dev. 14 (2000) 627). Recently, the transmembrane protein BAMBI (BMP and activin membrane-bound inhibitor) has been shown to interfere with Bmp and activin-like signaling by inhibiting Tgf-beta type I receptor activation (Nature 401 (1999) 480). In striking contrast to other Bmp antagonists like noggin (Cell 86 (1996) 599) or chordin (Cell 86 (1996) 589), BAMBI is strictly coexpressed with Bmp-4 during early Xenopus embryogenesis. The grouping of genes according to their shared complex spatial expression pattern and their involvement in the same biological signaling pathway has been referred to as synexpression group. This concept facilitates prognoses about the roles of a group member with unknown function. Apparently, only a minority of genes is organized in synexpression groups and up to now they have mainly been described in yeast and Xenopus (for review see Nature 402 (1999) 483). In the frog, BAMBI is a member of the Bmp-4 synexpression group (Nature 401 (1999) 480). We identified two murine homologues of BAMBI one of which, named Bambi-psi, is a pseudogene. We show that the spatiotemporal expression pattern of Bambi closely matches that of Bmp-4 during mouse embryonic development. Moreover, we show that Bambi expression is induced in mouse embryonic fibroblasts by Bmp-4. Hence, we provide first evidence for the existence of an evolutionarily conserved Bmp-4 synexpression group in mammals.
Asunto(s)
Proteínas Morfogenéticas Óseas/biosíntesis , Embrión de Mamíferos/metabolismo , Expresión Génica , Proteínas de la Membrana/biosíntesis , Proteínas de la Membrana/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteína Morfogenética Ósea 4 , Etiquetas de Secuencia Expresada , Extremidades/embriología , Humanos , Ratones , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Ácido Nucleico , Transducción de Señal , Factores de Tiempo , Distribución Tisular , Proteínas de XenopusRESUMEN
Mammalian homologues of the Drosophila Iroquois homeobox gene complex, involved in patterning and regionalization of differentiation, have recently been identified (Mech. Dev., 69 (1997) 169; Dev. Biol., 217 (2000) 266; Dev. Dyn., 218 (2000) 160; Mech. Dev., 91 (2000) 317; Dev. Biol., 224 (2000) 263; Genome Res., 10 (2000) 1453; Mech. Dev., 103 (2001) 193). The six members of the murine family were found to be organized in two cognate clusters of three genes each, Irx1, -2, -4 and Irx3, -5, -6, respectively (Peters et al., 2000). As a basis for further study of their regulation and function we performed a comparative analysis of the genomic organization and of the expression patterns of all six Irx genes. The genes are expressed in highly specific and regionalized patterns of ectoderm, mesoderm and endoderm derived tissues. In most tissues the pattern of expression of the clustered genes, especially of Irx1 and -2 and of Irx3 and -5, respectively, closely resembled each other while those of Irx4 and -6 were very divergent. Interestingly, the expression of cognate genes was found to be mutually exclusive in adjacent and interacting tissues of limb, heart and the laryncho-pharyncheal region. The results indicate that the Irx genes are coordinately regulated at the level of the cluster.
Asunto(s)
Embrión de Mamíferos/metabolismo , Proteínas de Homeodominio/genética , Familia de Multigenes , Factores de Transcripción/genética , Animales , Sistema Nervioso Central/embriología , Sistema Digestivo/embriología , Desarrollo Embrionario y Fetal , Epidermis/embriología , Epidermis/metabolismo , Extremidades/embriología , Perfilación de la Expresión Génica , Gónadas/embriología , Gónadas/metabolismo , Corazón/embriología , Hibridación in Situ , Riñón/embriología , Riñón/metabolismo , Ratones , Sistema Respiratorio/embriología , Sistema Respiratorio/metabolismoRESUMEN
Iroquois (Irx) proteins comprise a family of homeodomain-containing transcription factors involved in patterning and regionalization of embryonic tissues in both vertebrates and invertebrates. The six murine Irx genes are organized in two clusters, each consisting of three genes. Irx1, Irx2 and Irx4 belong to the IrxA cluster on chromosome 13, whereas Irx3, Irx5 and Irx6, comprising the IrxB cluster, are located on chromosome 8 (Peters et al., Genome Res. 10 (2000) 1453). Developmental expression patterns have so far only been reported for five Irx genes. Here, we investigated the expression pattern of Irx6 during mouse morphogenesis and early organogenesis. The pattern was found to be much more restricted compared to the other five Irx genes, and its level of expression was much lower.