Journal
EXPERIMENTAL CELL RESEARCH
Volume 303, Issue 2, Pages 252-262Publisher
ELSEVIER INC
DOI: 10.1016/j.yexcr.2004.10.006
Keywords
intestine; development; apoptosis; proliferation; organ culture; caspase-3; epithelial; EGF; AG1478
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Differentiation of endoderm into intestinal epithelium is initiated at E 13.5 of mouse development when there are significant changes in morphology resulting in the conversion of undifferentiated stratified epithelium into a mature epithelial monolayer. Here we demonstrate that monolayer formation is associated with the selective apoptosis of superficial cells lining the lumen while cell proliferation is progressively restricted to cells adjacent to the basement membrane. We describe an innovative embryonic gut culture system that maintains the three-dimensional architecture of gut and in which these processes are recapitulated in vitro. Explants taken from specific regions of the gut and placed into organ culture develop and express molecular markers (Cdx1, Cdx2 and A33 antigen) in the same spatial and temporal pattern observed in vivo indicating that regional specification is maintained. Inhibition of the epidermal growth factor receptor (EGFR) tyrosine kinase using the specific inhibitor AG1478 significantly reduced the proliferation and survival of cells within the epithelial cell layer of cultured gut explants. This demonstrates an essential role for the EGF signalling pathway during the early stages of intestinal development. (C) 2004 Elsevier Inc. All rights reserved.
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