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Isolation of the Xenopus homolog of int-1/wingless and expression during neurula stages of early development. 总被引:6,自引:2,他引:4
We have isolated the Xenopus homolog (Xint-1) of the mouse protooncogene int-1 from a neurula stage 17 cDNA library. The deduced protein sequence of Xint-1 includes 371 amino acids. The Xint-1 protein is more similar to the mammalian int-1 product (69%), than to the Drosophila counterpart of int-1, wingless (50%). Xint-1 shares several characteristics of secreted proteins with the other int-1 homologs: it has a hydrophobic leader, multiple conserved potential N-linked glycosylation sites and is rich in cysteine residues. All 23 cysteines are conserved in the three proteins. Xint-1 is transiently expressed during the neurula stages of early Xenopus development. 相似文献
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Developmental expression of the Xenopus int-2 (FGF-3) gene: activation by mesodermal and neural induction. 总被引:2,自引:0,他引:2
D Tannahill H V Isaacs M J Close G Peters J M Slack 《Development (Cambridge, England)》1992,115(3):695-702
We have used a probe specific for the Xenopus homologue of the mammalian proto-oncogene int-2 (FGF-3) to examine the temporal and spatial expression pattern of the gene during Xenopus development. int-2 is expressed from just before the onset of gastrulation through to prelarval stages. In the early gastrula, it is expressed around the blastopore lip. This is maintained in the posterior third of the prospective mesoderm and neuroectoderm in the neurula. A second expression domain in the anterior third of the neuroectoderm alone appears in the late gastrula, which later resolves into the optic vesicles, hypothalamus and midbrain-hindbrain junction region. Further domains of expression arise in tailbud to prelarval embryos, including the stomodeal mesenchyme, the endoderm of the pharyngeal pouches and the cranial ganglia flanking the otocyst. It is shown, by treatment of blastula ectoderm with bFGF and activin, that int-2 can be expressed in response to mesoderm induction. By heterotypic grafting of gastrula ectoderm into axolotl neural plate, we have also demonstrated that int-2 can be expressed in response to neural induction. These results suggest that int-2 has multiple functions in development, including an early role in patterning of the anteroposterior body axis and a later role in the development of the tail, brain-derived structures and other epithelia. 相似文献
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Ectopic expression of the proto-oncogene int-1 in Xenopus embryos leads to duplication of the embryonic axis 总被引:24,自引:0,他引:24
While there is convincing evidence implicating Drosophila int-1 in pattern regulation, the normal role of int-1 in vertebrate development is unclear. We have injected Xenopus eggs with mouse int-1 RNA and monitored subsequent development. Injected RNA is translated and the protein widely distributed. Embryos develop into apparently normal gastrulae, but almost all surviving neurulae have a bifurcated anterior and expanded posterior neural plate. Bifurcation of the neural plate was abolished by substitution of a single, conserved cysteine residue and was dependent on the presence of a signal peptide sequence in the int-1 protein. Histological examination indicates that underlying axial mesodermal structures were duplicated. This result suggests that ectopic int-1 expression leads to dual axis formation and points to a role for int-1 in patterning processes in vertebrate development. 相似文献
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Expression of the proto-oncogene int-1 is restricted to specific neural cells in the developing mouse embryo 总被引:51,自引:0,他引:51
We have used in situ hybridization and computer-aided reconstruction to study the spatial distribution of expression of the mammary tumor proto-oncogene int-1 during mouse embryogenesis. int-1 RNA accumulation is restricted to specific regions of the neural plate and its derivatives between 9 and 14.5 days of development. int-1 RNA accumulates throughout the neural plate at the anterior head folds of the 9 day embryo but only at its lateral tips in more posterior regions. Following neural tube closure, int-1 expression is restricted to specific regions of the dorsal wall of the brain ventricles and spinal cord, the ventral wall of the midbrain and the diencephalon, and the lateral walls of the neuroepithelium at the midbrain-hindbrain junction. These data suggest that int-1 has a role in the early stages of central nervous system development in the mouse embryo. 相似文献
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The proto-oncogene int-1 encodes a secreted protein associated with the extracellular matrix. 总被引:32,自引:5,他引:27
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The proto-oncogene int-1 plays an important role in mammary tumorigenesis when activated by proviral insertions of the mouse mammary tumor virus. In normal mouse tissues the gene is expressed in the embryonic neural tube, suggesting a developmental function, while in Drosophila the homolog of int-1 is the segment polarity gene wingless. In order to study the protein products of int-1 we have derived fibroblast cell lines infected with multiple copies of a retroviral vector expressing int-1 cDNA. By Western blot analysis and immunoprecipitation we have identified a 44 kd form of int-1 protein which is secreted from these cells. The 44 kd species is distinct from the major intracellular forms of int-1 protein as judged by its slower mobility in SDS-polyacrylamide gels and by its longer half-life in pulse-chase experiments. Under normal growth conditions, little or none of the 44 kd protein is detectable in the cell culture medium but instead the majority is found associated with the extracellular matrix (ECM). The protein appears to bind heparin in vitro, suggesting that it might bind glycosaminoglycans in the ECM. These data support the view that int-1 protein may play a role in cell-cell communication over short distances. 相似文献
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J Papkoff 《Molecular and cellular biology》1989,9(8):3377-3384
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A M Brown J Papkoff Y K Fung G M Shackleford H E Varmus 《Molecular and cellular biology》1987,7(11):3971-3977
The proto-oncogene int-1 is activated by adjacent insertions of proviral DNA in mouse mammary tumor virus-induced tumors and has transforming activity in certain mammary epithelial cell lines. The gene is normally expressed in the central nervous system of mid-gestational embryos and in the adult testis. We raised antibodies against synthetic int-1 peptides and used these to identify protein products of the gene in cells transfected or infected with retroviral vectors expressing int-1. Four protein species of 36,000, 38,000, 40,000, and 42,000 Mr were immunoprecipitated by antibodies against two different int-1 peptides and were not present in control cells. Partial degradation with V8 protease showed the four species to be structurally related to each other and to int-1 polypeptide synthesized in vitro. Treatment of the cells with tunicamycin prevented the appearance of all but the 36,000-Mr species, suggesting that the slower-migrating forms are glycosylated derivatives. The unglycosylated 36,000-Mr species migrated faster in polyacrylamide gels than the in vitro translation product of int-1 and has probably undergone cleavage of an amino-terminal signal peptide. 相似文献
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Wang C Xia C Bian W Liu L Lin W Chen YG Ang SL Jing N 《Molecular biology of the cell》2006,17(7):3075-3084
FGF8, a member of the fibroblast growth factor (FGF) family, has been shown to play important roles in different developing systems. Mouse embryonic carcinoma P19 cells could be induced by retinoic acid (RA) to differentiate into neuroectodermal cell lineages, and this process is cell aggregation dependent. In this report, we show that FGF8 expression is transiently up-regulated upon P19 cell aggregation, and the aggregation-dependent FGF8 elevation is pluripotent stem cell related. Overexpressing FGF8 promotes RA-induced monolayer P19 cell neural differentiation. Inhibition of FGF8 expression by RNA interference or blocking FGF signaling by the FGF receptor inhibitor, SU5402, attenuates neural differentiation of the P19 cell. Blocking the bone morphogenetic protein (BMP) pathway by overexpressing Smad6 in P19 cells, we also show that FGF signaling plays a BMP inhibition-independent role in P19 cell neural differentiation. 相似文献
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T Venesio D Taverna N E Hynes R Deed D MacAllan F Ciardiello E M Valverius D S Salomon R Callahan G Merlo 《Cell growth & differentiation》1992,3(1):63-71
We have investigated the effect of basic fibroblast growth factor (bFGF) and the related int-2 gene on the growth, transformation, and differentiation of HC11 mouse mammary epithelial cells. We show that in HC11 cells infected with int-2 retroviral expression vectors, the int-2 protein can function as a bFGF-like growth factor in stimulating: (a) HC11 cell proliferation in monolayer, (b) anchorage-independent growth in soft agar, and (c) soft agar growth of the bFGF-responsive SW13 tumor cell line. These effects are observed irrespective of whether the int-2 protein is expressed in its wild-type form or is linked to a signal peptide. A candidate bFGF receptor, which is the product of the flg gene and which may recognize the int-2 protein, is expressed at high levels in HC11 cells. Following epidermal growth factor or bFGF priming and subsequent treatment with lactogenic hormones, all of the int-2 infected and the parental HC11 cells synthesize similar levels of beta-casein. However, the autocrine expression of int-2 in HC11 cells abrogates their requirement for either exogenous epidermal growth factor or bFGF priming. These data suggest that, in HC11 cells, the growth factor activity of the int-2 gene is indistinguishable from that of bFGF and does not interfere with the mammary cell differentiation program associated with lactogenesis. 相似文献
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Fgf8 is a member of the fibroblast growth factor (FGF) family that plays an important role in early neural development. Cellular aggregation and retinoic acid (RA) are needed for mouse embryonic carcinoma (EC) P19 cell neural differentiation. We have examined the Fgf8 gene in P19 cells during neural differentiation and identified 2 alternatively spliced Fgf8 isoforms, Fgf8a and Fgf8b, among the 8 known splicing isoforms in mammals. The expression of Fgf8a and Fgf8b mRNAs transiently and rapidly increased in the early stage of P19 cells during RA-induced neural differentiation, followed by a decline in expression. The relative amount of Fgf8b was clearly higher than that of Fgf8a at different time-points measured within 24 h after RA treatment. Increased Fgf8b mRNA expression was cellular-aggregation dependent. The results demonstrated that cellular-aggregation-induced Fgf8b, but not Fgf8a, may play a pivotal role in early neural differentiation of P19 cells. 相似文献