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An investigation of endoderm to mesoderm signalling in gut development

By Emma Jane Stringer


It has been suggested that the intestinal endoderm is responsive to signals received\ud from underlying mesoderm, indicating cross-talk between the layers. Cdx2 mutant\ud mice develop heterotopias of stomach-type epithelium within the paracaecal region of\ud the intestine. They are therefore an ideal in-vivo model in which to study whether the\ud loss of endodermal gene expression required for intestinal development leads to\ud stomach-specific mesodermal gene expression.\ud In this study, Sox2, a stomach endoderm-specific marker, was used to detect gastric\ud heterotopias in Cdx2 mutant embryonic intestine using whole-mount in-situ\ud hybridisation. The nature of the underlying mesoderm was investigated using a\ud second marker, Barx1, which is known to be specifically expressed in the stomach\ud mesoderm. RT-PCR was used to detect low levels of Barx1 expression in Cdx2+/-\ud caecum samples. Further investigation using in-situ hybridisation techniques\ud indicated regions of Barx1 expression in a similar distribution to the regions detected\ud using the Sox2 probe. This finding confirms that the mesoderm underlying the Cdx2\ud mutant gastric heterotopias expresses a stomach-specific gene and therefore that\ud the mesoderm is responsive to endodermal signals.\ud It has been suggested that Cdx2+/- mice do not develop gastric-type intestinal\ud heterotopias postnatally. If proven, this would indicate that intestinal stem cell\ud potential becomes limited at some point during development and prevents the\ud epithelium responding to a postnatal loss of Cdx2 protein. A conditional Cdx2 mouse\ud model is required in order to investigate this hypothesis. The creation of this mouse\ud model formed the second part of this project. Following creation of a targeting vector,\ud transfection into ES cells and injection into blastocysts, chimeric mice were obtained.\ud These mice were successfully bred for germline transmission

Publisher: University of Leicester
Year: 2008
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