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An investigation of endoderm to mesoderm signalling in gut development

By Emma Jane Stringer

Abstract

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
OAI identifier: oai:lra.le.ac.uk:2381/9919

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Citations

  1. (1988). A clonal marker induced by mutation in mouse intestinal epithelium.
  2. (1994). A homeodomain protein related to caudal regulates intestine-specific gene transcription.
  3. (1995). A member of the caudal family of homeobox genes maps to the X-inactivation centre region of the mouse and human X chromosomes.
  4. (1995). A modified oestrogen receptor ligand-binding domain as an improved switch for the regulation of heterologous proteins.
  5. (1988). A mouse gene homologous to the Drosophila gene caudal is expressed in epithelial cells from the embryonic intestine.
  6. (1992). A multipoint genetic linkage map of mouse chromosome 18.
  7. (2007). a murine model of intestinal inflammation.
  8. (1994). A primary requirement for nodal in the formation and maintenance of the primitive streak in the mouse.
  9. (1993). A rational nomenclature for vertebrate homeobox (HOX) genes.
  10. (2003). A study of regional gut endoderm potency by analysis of Cdx2 null mutant chimaeric mice.
  11. (2004). A transgenic mouse model with inducible Tyrosinase gene expression using the tetracycline (Tet-on) system allows regulated rescue of abnormal chiasmatic projections found in albinism.
  12. (1997). Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC.
  13. (1997). Activation of enhancer elements by the homeobox gene Cdx2 is cell line specific.
  14. (1998). Age changes in stem cells of murine small intestinal crypts.
  15. (1998). Altered stem cell regeneration in irradiated intestinal crypts of senescent mice.
  16. (1998). An Fgf8 mutant allelic series generated by Cre- and Flp-mediated recombination.
  17. (1996). Anterior primitive endoderm may be responsible for patterning the anterior neural plate in the mouse embryo.
  18. (2008). Appendix of the midgut endoderm’,
  19. (1999). Axis development and early asymmetry in mammals.
  20. (1995). Barx1 expression can be detected in the midgut of embryos expressing ectopic Sox2 Since Sox2 expressing areas of heterotopia were detected in Cdx2+/ and Cdx2///WT embryos, the nature of the gut mesoderm was investigated.
  21. (1995). Barx1, a new mouse homeodomain transcription factor expressed in cranio-facial ectomesenchyme and the stomach.
  22. (2002). Beta-catenin and TCF mediate cell positioning in the intestinal epithelium by controlling the expression of EphB/ephrinB.
  23. (1986). Bloom + Fawcett, A Textbook of Histology.
  24. (2008). Bmi1 is expressed in vivo in intestinal stem cells.
  25. (1999). BMP signalling specifies the pyloric sphincter.
  26. (1999). Bone marrow as a potential source of hepatic oval cells.
  27. (2005). C/EBP and Cdx family factors regulate liver fatty acid binding protein transgene expression in the small intestinal epithelium.
  28. (2003). Canonical Wnt signals are essential for homeostasis of the intestinal epithelium.
  29. (2005). Cdk2-dependent phosphorylation of homeobox transcription factor CDX2 regulates its nuclear translocation and proteasome-mediated degradation in human intestinal epithelial cells.
  30. (2006). Cdx-2 expression in squamous and metaplastic columnar epithelia of the esophagus.
  31. (1995). Cdx-2 homeodomain protein expression in human and rat colorectal adenoma and carcinoma.
  32. (2004). Cdx1 induced intestinal metaplasia in the transgenic mouse stomach: comparative study with Cdx2 transgenic mice.
  33. (2002). Cdx2 ectopic expression induces gastric intestinal metaplasia in transgenic mice.
  34. (2004). Cdx2 is essential for axial elongation in mouse development.
  35. (2005). Cdx2 is required for correct cell fate specification and differentiation of trophectoderm in the mouse blastocyst.
  36. (2006). Cdx2 modulates proliferation in normal human intestinal epithelial crypt cells.
  37. (1987). Cell fate, morphogenetic movement and population kinetics of embryonic endoderm at the time of germ layer formation in the mouse.
  38. (1999). Cell type-specific autoregulation of the Caudal-related homeobox gene Cdx-2/3.
  39. (1998). Cellular and molecular partners involved in gut morphogenesis and differentiation.
  40. (2005). Cellular and molecular regulation of hematopoietic and intestinal stem cell behavior.
  41. (1996). Cerberus is a head-inducing secreted factor expressed in the anterior endoderm of Spemann's organizer.
  42. (2005). Characterization of mesendoderm: a diverging point of the definitive endoderm and mesoderm in embryonic stem cell differentiation culture.
  43. (1996). Chromosomal localization of cdx2, a murine homologue of the Drosophila gene caudal, to mouse chromosome 5.
  44. (1996). cis-Acting elements and transcription factors involved in the intestinal specific expression of the rat calbindin-D9K gene: binding of the intestinespecific transcription factor Cdx-2 to the TATA box.
  45. (1999). Clonal analysis of mouse intestinal epithelial progenitors.
  46. (1997). Clonality and life cycles of intestinal crypts explained by a state dependent stochastic model of epithelial stem cell organization.
  47. (2003). Colonic polyposis caused by mTOR-mediated chromosomal instability in Apc+/Delta716 Cdx2+/- compound mutant mice.
  48. (2000). Color Atlas of Histology. Lippincott Williams and Wilkins, Third Edition.
  49. (2000). Control of endodermal endocrine development by Hes-1.
  50. (2002). Conversion of gastric mucosa to intestinal metaplasia in Cdx2-expressing transgenic mice.
  51. Correlation with attenuation of apoptosis in colonic crypts and the incidence of colonic neoplasia.
  52. (2000). Cre recombinase: the universal reagent for genome tailoring.
  53. (1999). CREB-binding [corrected] protein interacts with the homeodomain protein Cdx2 and enhances transcriptional activity.
  54. (2005). Crypt-restricted proliferation and commitment to the Paneth cell lineage following Apc loss in the mouse intestine.
  55. (2001). Defects of the body plan of mutant embryos lacking Lim1, Otx2 or Hnf3beta activity.
  56. (2006). Definitive endoderm of the mouse embryo: formation, cell fates, and morphogenetic function.
  57. (1998). Depletion of epithelial stem-cell compartments in the small intestine of mice lacking Tcf-4.
  58. (2004). Development of gastric carcinoma from intestinal metaplasia in Cdx2-transgenic mice.
  59. (1988). Development of the pattern of cell renewal in the crypt-villus unit of chimaeric mouse small intestine.
  60. (1990). Development of the vertebrate small intestine and mechanisms of cell differentiation.
  61. (2005). Developmental regulation of the Hox genes
  62. (2001). Differential activation of intestinal gene promoters: functional interactions between GATA-5 and HNF-1 alpha.
  63. (1995). Differential expression of bcl-2 in intestinal epithelia.
  64. (1997). Disruption of overlapping transcripts in the ROSA beta geo 26 gene trap strain leads to widespread expression of betagalactosidase in mouse embryos and hematopoietic cells.
  65. (1995). Disruption of the murine homeobox gene Cdx1 affects axial skeletal identities by altering the mesodermal expression domains of Hox genes.
  66. (1986). DNAmediated gene transfer into epidermal cells using electroporation.
  67. (2000). Early mouse endoderm is patterned by soluble factors from adjacent germ layers.
  68. (2002). Efficient recombination in diverse tissues by a tamoxifen-inducible form of Cre: a tool for temporally regulated gene activation/inactivation in the mouse.
  69. (1996). eFGF, Xcad3 and Hox genes form a molecular pathway that establishes the anteroposterior axis in Xenopus.
  70. (2004). Elimination of background recombination: somatic induction of Cre by combined transcriptional regulation and hormone binding affinity.
  71. (2001). Embryonic expression of an Nkx2-5/Cre gene using ROSA26 reporter mice.
  72. (1998). Embryonic stem cell lines derived from human blastocysts.
  73. (1996). Epithelial cell migration in the intestine.
  74. (1998). Epithelialmesenchymal signaling during the regionalization of the chick gut.
  75. (1981). Establishment in culture of pluripotential cells from mouse embryos.
  76. (2000). Evidence for 14 homeobox gene clusters in human genome ancestry.
  77. (2000). Evolutionary relationships between the amphibian, avian, and mammalian stomachs.
  78. (1995). Expression of Cdx-2 in the mouse embryo and placenta: possible role in patterning of the extraembryonic membranes.
  79. (0014). Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
  80. (2006). FGF signal transduction and the regulation of Cdx gene expression.
  81. (2006). FGF signaling is necessary for establishing gut tube domains along the anterior–posterior axis in vivo.
  82. (1999). Fgf-8 determines rostral-caudal polarity in the first branchial arch.
  83. (1996). Flp recombinase promotes site-specific DNA recombination in embryonic stem cells and transgenic mice.
  84. (1984). Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines.
  85. (2007). Frequent rearrangements and amplification of the CDX2 homeobox gene in human sporadic colorectal cancers with chromosomal instability.
  86. (1997). Gastric epithelial morphogenesis in normal and transgenic mice.
  87. (2006). Gata4 is essential for the maintenance of jejunal-ileal identities in the adult mouse small intestine.
  88. (2007). Gene expression patterns of human colon tops and basal crypts and BMP antagonists as intestinal stem cell niche factors.
  89. (1999). Generalized lacZ expression with the ROSA26 Cre reporter strain.
  90. (2001). Genetic dissection of nodal function in patterning the mouse embryo.
  91. (2000). Gut instincts: thoughts on intestinal epithelial stem cells.
  92. (1997). Hematopoietic cells differentiate into both microglia and macroglia in the brains of adult mice.
  93. (2000). High-efficiency deleter mice show that FLPe is an alternative to Cre-loxP.
  94. (2004). Hindgut defects and transformation of the gastro-intestinal tract in Tcf4(-/-)/Tcf1(-/-) embryos.
  95. (2004). Hindgut defects and transformation of the gastro-intestinal tract in Tcf4(/)/Tcf1(/) embryos.
  96. (1996). Hlx homeo box gene is essential for an inductive tissue interaction that drives expansion of embryonic liver and gut.
  97. (1994). HNF-3 beta is essential for node and notochord formation in mouse development.
  98. (1991). Homeobox gene expression in the intestinal epithelium of adult mice.
  99. (1992). Homeobox genes and axial patterning.
  100. (2000). Homeobox genes and gut development.
  101. (2002). Homeobox genes in gut development.
  102. (1997). Homeosis and intestinal tumours
  103. (1997). Homeosis and intestinal tumours in Cdx2 mutant mice.
  104. (1995). Homology requirements for ligation and strand exchange by the FLP recombinase.
  105. (1985). Human Embryology. Blackwell Scientific Publications, Second Edition.
  106. (1993). Identification of residues in the estrogen receptor that confer differential sensitivity to estrogen and hydroxytamoxifen.
  107. (2007). Identification of stem cells in small intestine and colon by marker gene Lgr5.
  108. (1986). Identification of the crossover site during FLP-mediated recombination in the Saccharomyces cerevisiae plasmid 2 microns circle.
  109. (2004). Identifying target genes regulated downstream of Cdx2 by microarray analysis.
  110. (1996). Improved properties of FLP recombinase evolved by cycling mutagenesis.
  111. (1992). In Situ Hybridisation: A Practical Approach,
  112. (2002). Inactivation of the transcription factor Elf3 in mice results in dysmorphogenesis and altered differentiation of intestinal epithelium.
  113. (2007). Independent functions and mechanisms for homeobox gene
  114. (2007). Independent functions and mechanisms for homeobox gene Barx1 in patterning mouse stomach and spleen.
  115. (2004). Inducible Cre-mediated control of gene expression in the murine gastrointestinal tract: effect of loss of beta-catenin.
  116. (1999). Inducible gene knockouts in the small intestinal and colonic epithelium.
  117. (2002). Inhibition of excess nodal signaling during mouse gastrulation by the transcriptional corepressor DRAP1.
  118. (1988). Inhibition of pluripotential embryonic stem cell differentiation by purified polypeptides.
  119. (2002). Initiating Hox gene expression: in the early chick neural tube differential sensitivity to FGF and RA signaling subdivides the HoxB genes in two distinct groups.
  120. (2000). Interaction between the homeodomain proteins Cdx2 and HNF1alpha mediates expression of the lactase-phlorizin hydrolase gene.
  121. (1984). Interaction of the bacteriophage P1 recombinase Cre with the recombining site loxP.
  122. (2001). Intestinal epithelial cell differentiation involves activation of p38 mitogenactivated protein kinase that regulates the homeobox transcription factor CDX2.
  123. (1998). Intestinal epithelial-mesenchymal cell interactions.
  124. (1998). Intestinal epithelial–mesenchymal cell interactions.
  125. (1998). Intestinal expression of the calbindin-D9K gene in transgenic mice. Requirement for a Cdx2-binding site in a distal activator region.
  126. (2002). Intestinal stem cells protect their genome by selective segregation of template DNA strands.
  127. (1981). Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells.
  128. (1997). Key role of the Cdx2 homeobox gene in extracellular matrix-mediated intestinal cell differentiation.
  129. (1997). Laboratory Protocols for Conditional Gene Targeting,
  130. (1996). Ligand-activated site-specific recombination in mice.
  131. (2000). Linking colorectal cancer to Wnt signaling.
  132. (2003). Live and let die in the intestinal epithelium.
  133. (1996). Mesodermspecific expression of the divergent homeobox gene Hlx during murine embryogenesis.
  134. (1998). Modification of gene activity in mouse embryos in utero by a tamoxifen-inducible form of Cre recombinase.
  135. (1993). Mouse Cdx-1 expression during gastrulation.
  136. (2005). Mtgr1 is a transcriptional corepressor that is required for maintenance of the secretory cell lineage in the small intestine.
  137. (1999). Multilineage potential of adult human mesenchymal stem cells.
  138. (2007). Multiple dose-dependent roles for Sox2 in the patterning and differentiation of anterior foregut endoderm.
  139. (2003). Multipotent cell lineages in early mouse development depend on SOX2 function.
  140. (1993). Murine Cdx-4 bears striking similarities to the Drosophila caudal gene in its homeodomain sequence and early expression pattern.
  141. (1998). Muscle regeneration by bone marrow-derived myogenic progenitors.
  142. (2002). Neurogenin3 is differentially required for endocrine cell fate specification in the intestinal and gastric epithelium.
  143. (1998). new roles in regulation of early hindbrain patterning.
  144. (1996). NK-2 homeobox genes and heart development.
  145. (1993). Nkx-2.5: a novel murine homeobox gene expressed in early heart progenitor cells and their myogenic descendants.
  146. (2005). No organ left behind: tales of gut development and evolution.
  147. (2005). Notch signals control the fate of immature progenitor cells in the intestine.
  148. (2005). Notch/gamma-secretase inhibition turns proliferative cells in intestinal crypts and adenomas into goblet cells.
  149. (1971). Observations on the structure of diphtheria toxin.
  150. (1998). Opposite phenotypes of hypomorphic and Y766 phosphorylation site mutations reveal a function for Fgfr1 in anteroposterior patterning of mouse embryos.
  151. (2005). Overlapping gene expression in fetal mouse intestine development and human colorectal cancer.
  152. (1982). P1 site-specific recombination: nucleotide sequence of the recombining sites.
  153. (1988). Pepsinogen gene transcription induced in heterologous epithelial-mesenchymal recombinations of chicken endoderms and glandular stomach mesenchyme.
  154. (2007). Phases of canonical Wnt signaling during the development of mouse intestinal epithelium.
  155. (2005). Phosphorylation of the homeotic tumor suppressor Cdx2 mediates its ubiquitin-dependent proteasome degradation.
  156. (2001). Phosphorylation of the serine 60 residue within the Cdx2 activation domain mediates its transactivation capacity.
  157. (2005). Plasticity of cultured mesenchymal stem cells: switch from nestin-positive to excitable neuron-like phenotype.
  158. (1981). Postnatal development: coordination of feeding, digestion, and metabolism.
  159. (2001). Pou homeodomain protein OCT1 is implicated in the expression of the caudal-related homeobox gene Cdx-2.
  160. (1998). Region-specific expression of chicken Sox2 in the developing gut and lung epithelium: regulation by epithelial-mesenchymal interactions.
  161. (1998). Region-specific expression of murine Hox genes implies the Hox code-mediated patterning of the digestive tract.
  162. (2002). Region-specific gastrointestinal Hox code during murine embryonal gut development.
  163. (1997). Regulation of lactase-phlorizin hydrolase gene expression by the caudal-related homoeodomain protein Cdx-2.
  164. (1994). Regulation of protein function through expression of chimaeric proteins.
  165. (1999). Reprogramming of intestinal differentiation and intercalary regeneration in Cdx2 mutant mice.
  166. (2007). required for the expression of specific intestinal genes during development.
  167. (1999). Requirement for Wnt3 in vertebrate axis formation.
  168. (2001). Requirement of Math1 for secretory cell lineage commitment in the mouse intestine.
  169. (2006). Role for retinoid signaling in left-right asymmetric digestive organ morphogenesis.
  170. (2000). Roles of BMP signaling and Nkx2.5 in patterning at the chick midgut-foregut boundary.
  171. (2000). Secretion of microbicidal alpha-defensins by intestinal Paneth cells in response to bacteria.
  172. (2005). Self-renewal and cancer of the gut: two sides of a coin.
  173. (2007). Sequential allocation and global pattern of movement of the definitive endoderm in the mouse embryo during gastrulation.
  174. (2000). Skin and hair follicle integrity is crucially dependent on beta 1 integrin expression on keratinocytes.
  175. (1995). Sonic hedgehog is an endodermal signal inducing Bmp4 and Hox genes during induction and regionalization of the chick hindgut.
  176. (2004). SOX9 is an intestine crypt transcription factor, is regulated by the Wnt pathway, and represses the CDX2 and MUC2 genes.
  177. (2007). SOX9 is required for the differentiation of paneth cells in the intestinal epithelium.
  178. (1998). Spatial pattern of smooth muscle differentiation is specified by the epithelium in the stomach of mouse embryo.
  179. (2006). Spatio-temporal patterns of intestinespecific transcription factor expression during postnatal mouse gut development.
  180. (1990). Specific expression of lactase in the jejunum and colon during postnatal development and hormone treatments in the rat.
  181. (1998). Stem cells in gastrointestinal epithelium: numbers, characteristics and death.
  182. (1990). Stem cells: attributes, cycles, spirals, pitfalls and uncertainties. Lessons for and from the crypt.
  183. (1971). Structure and activity of diphtheria toxin.
  184. (1994). Structure of the murine homeobox gene cdx-2. Expression in embryonic and adult intestinal epithelium.
  185. (1999). Targeted disruption of the homeobox transcription factor Nkx2-3 in mice results in postnatal lethality and abnormal development of small intestine and spleen.
  186. (2000). Temporospatial cell interactions regulating mandibular and maxillary arch patterning.
  187. (1999). The Anatomical Basis of Mouse Development,
  188. (2002). The beta-catenin/TCF-4 complex imposes a crypt progenitor phenotype on colorectal cancer cells.
  189. (1999). The caudal-related homeodomain protein Cdx-2 regulates vitamin D receptor gene expression in the small intestine.
  190. (2003). The Cdx2 homeobox gene has a tumour suppressor function in the distal colon in addition to a homeotic role during gut development.
  191. (2006). The Cdx4 mutation affects axial development and reveals an essential role of Cdx genes in the ontogenesis of the placental labyrinth in mice.
  192. (1994). The crypt cycle in mouse small intestinal epithelium.
  193. (2004). The cutting-edge of mammalian development; how the embryo makes teeth.
  194. (1987). The formation of mesodermal tissues in the mouse embryo during gastrulation and early organogenesis.
  195. (2004). The gastrointestinal stem cell.
  196. (2004). The homeobox intestinal differentiation factor CDX2 is selectively expressed in gastrointestinal adenocarcinomas.
  197. (1997). The intestinal epithelial stem cell: the mucosal governor.
  198. (2002). The intestinal epithelial stem cell.
  199. (2007). The intestinespecific homeobox gene Cdx2 decreases mobility and antagonizes dissemination of colon cancer cells.
  200. (1997). The mesenchymal winged helix transcription factor Fkh6 is required for the control of gastrointestinal proliferation and differentiation.
  201. (1997). The mouse Nkx2-3 homeodomain gene is expressed in gut mesenchyme during pre- and postnatal mouse development.
  202. (1998). The murine Cdx1 gene product localises to the proliferative compartment in the developing and regenerating intestinal epithelium.
  203. (1989). The response of murine intestinal crypts to shortrange promethium-147 beta irradiation: deductions concerning clonogenic cell numbers and positions.
  204. (1994). The role of p53 in spontaneous and radiation-induced apoptosis in the gastrointestinal tract of normal and p53-deficient mice.
  205. (1978). The segregation of DNA in epithelial stem cells.
  206. (2001). The small musclespecific protein Csl modifies cell shape and promotes myocyte fusion in an insulin-like growth factor 1-dependent manner.
  207. (1981). The stem-cell zone of the small intestinal epithelium. III. Evidence from columnar, enteroendocrine, and mucous cells in the adult mouse.
  208. (2005). The stomach mesenchymal transcription factor Barx1 specifies gastric epithelial identity through inhibition of transient Wnt signaling.
  209. (1989). The structure and function of the homeodomain.
  210. (2002). The zinc-finger transcription factor Klf4 is required for terminal differentiation of goblet cells in the colon.
  211. Thiol-dependent dissociation of a fraction of toxin into enzymically active and inactive fragments.
  212. (2004). Tissuespecific and inducible Cre-mediated recombination in the gut epithelium.
  213. (1998). Transformation of tooth type induced by inhibition of BMP signaling.
  214. (1999). Vertebrate endoderm development.
  215. (2000). Widespread recombinase expression using FLPeR (flipper) mice.
  216. (2002). Wnt signaling controls the phosphorylation status of beta-catenin.
  217. (2001). Wnt-3a is required for somite specification along the anteroposterior axis of the mouse embryo and for regulation of cdx-1 expression.
  218. (2000). Wnt/(beta)-catenin signaling regulates the expression of the homeobox gene Cdx1 in embryonic intestine.
  219. (2003). Wnt3a plays a major role in the segmentation clock controlling somitogenesis.
  220. (2000). Z/EG, a double reporter mouse line that expresses enhanced green fluorescent protein upon Cre-mediated excision.

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