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J Exp Zool. 1984 Nov;232(2):287-96.

The role of cell proliferation and cellular shape change in branching morphogenesis of the embryonic mouse lung: analysis using aphidicolin and cytochalasins.


The formation of induced supernumerary buds in the embryonic mouse tracheal epithelium has been used as a model system to analyse the respective roles of cell proliferation and microfilament-mediated cell shape change during branching morphogenesis. In order to analyse the mitotic events associated with the formation of epithelial buds, the induction of supernumerary tracheal buds by mesenchymal grafts was carried out with the inhibitor of DNA synthesis, aphidicolin, present in the culture medium for varying intervals of time during the 16-hour inductive process. The presence of aphidicolin for 10 to 16 hours of the inductive period blocks the formation of induced tracheal buds, whereas the presence of the inhibitor for half of that time (either the first 8 hours or the last 8 hours) does not prevent this morphogenetic event from taking place, although smaller buds resulted from induction under these conditions. Both the inhibition of DNA synthesis and the recovery from 10 microM aphidicolin treatment, as measured by 3H-thymidine incorporation, were found to occur rapidly. The addition of 2 microM dihydrocytochalasin B (or cytochalasin B) together with aphidicolin during the second half of the inductive period inhibits the formation of supernumerary buds and upon removal of the cytochalasin rapid formation of buds takes place. We conclude that the formation of epithelial buds during branching morphogenesis occurs as a result of enhanced localized cell proliferation coupled with epithelial cell shape change (or preservation of cell morphology) mediated by microfilaments, which have been observed in both the apical and basal cytoplasm of the epithelial cells in the region where branching of the trachea is taking place.

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