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- Identification and functional analysis of novel genes expressed in the Anterior Visceral EndodermPublication . Gonçalves Dias Da Silva, Lisa; Filipe, Mario; Marques, Sara; Salgueiro, Ana Marisa; Becker, Jorg D.; Belo, José A.During early vertebrate development, the correct establishment of the body axes is critical. The anterior pole of the mouse embryo is established when Distal Visceral Endoderm (DVE) cells migrate to form the Anterior Visceral Endoderm (AVE). Symmetrical expression of Lefty1, Cer1 and Dkk1 determines the direction of DVE migration and the future anterior side. In addition to the establishment of the Anterior-Posterior axis, the AVE has also been implicated in anterior neural specification. To better understand the role of the AVE in these processes, we have performed a differential screening using Affymetrix GeneChip technology with AVE cells isolated from cer1P-EGFP transgenic mouse embryos. We found 175 genes which were upregulated in the AVE and 36 genes in the Proximal-posterior sample. Using DAVID software, we characterized the AVE cell population regarding cellular component, molecular function and biological processes. Among the genes that were found to be upregulated in the AVE, several novel genes were identified. Four of these transcripts displaying high-fold change in the AVE were further characterized by in situ hybridization in early stages of development in order to validate the screening. From those four selected genes, one, denominated Adtk1, was chosen to be functionally characterized by targeted inactivation in ES cells. Adtk1 encodes for a serine/threonine kinase. Adtk1 null mutants are smaller and present short limbs due to decreased mineralization, suggesting a potential role in chondrogenesis during limb development. Taken together, these data point to the importance of reporting novel genes present in the AVE.
- Comparative expression of mouse and chicken shisa homologues during early developmentPublication . Filipe, Mario; Gonçalves Dias da Silva, Lisa; Bento, Margaret; Silva, Ana Cristina; Belo, José A.During vertebrate embryogenesis, fibroblast growth factor (FGF) and Wnt signaling have been implicated in diverse cellular processes, including cell growth, differentiation, and tissue patterning. The recently identified Xenopus Shisa protein promotes head formation by inhibiting Wnt and FGF signaling through its interaction with the immature forms of Frizzled and FGF receptors in the endoplasmic reticulum, which prevents their posttranslational maturation. Here, we describe the mouse and chicken homologues of Xenopus Shisa. The mouse and chicken Shisa proteins share, respectively, 33.6% and 33.8% identity with the Xenopus homolog. In situ hybridization analysis shows that mouse shisa is expressed throughout embryonic development, predominantly in the anterior visceral endoderm, headfolds, somites, forebrain, optic vesicle, and limb buds. Cross-species comparison shows that the expression pattern of cshisa closely mirrors that of mshisa. Our observations indicate that the Shisa family genes are typically expressed in tissues known to require the modulation of Wnt and FGF signaling. Developmental Dynamics 235:2567-2573, 2006. (c) 2006 Wiley-Liss, Inc.
- Cripto-independent Nodal signaling promotes positioning of the A-P axis in the early mouse embryoPublication . Liguori, Giovanna; Borges, Ana; D'Andrea, Daniela; Liguoro, Annamaria; Gonçalves Dias da Silva, Lisa; Salizueiro, Ana Marisa; Persico, M. Graziella; Belo, José A.During early mouse development, the TGF beta-related protein Nodal specifies the organizing centers that control the formation of the anterior-posterior (A-P) axis. EGF-CFC proteins are important components of the Nodal signaling pathway, most likely by acting as Nodal coreceptors. However, the extent to which Nodal activity depends on EGF-CFC proteins is still debated. Cripto is the earliest EGF-CFC gene expressed during mouse embryogenesis and is involved in both A-P axis orientation and mesoderm formation. To investigate the relation between Cripto and Nodal in the early mouse embryo, we removed the Nodal antagonist Cerberus 1 (Cer1) and simultaneously Cripto, by generating Cer1;Cripto double mouse mutants. We observed that two thirds of the Cer1,Cripto double mutants are rescued in processes that are severely compromised in Cripto(-/-) embryos, namely A-P axis orientation, anterior mesendoderm and posterior neuroectoderin formation. The observed rescue is strongly reduced in Cer1;Cripto;Nodal triple mutants, suggesting that Nodal can signal extensively in the absence of Cripto, if Cer1 is also inhibited. This signaling activity drives A-P axis positioning. Our results provide evidence for the existence. of Cripto-independent signaling mechanisms, by which Nodal controls axis specification in the early mouse embryo. (C) 2007 Elsevier Inc. All rights reserved.