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Mitotic cell rounding accelerates epithelial invagination



Mitotic cells assume a spherical shape by increasing their surface tension and osmotic pressure by extensively reorganizing their interphase actin cytoskeleton into a cortical meshwork and their microtubules into the mitotic spindle1,2. Mitotic entry is known to interfere with tissue morphogenetic events that require cell-shape changes controlled by the interphase cytoskeleton, such as apical constriction3,4,5. However, here we show that mitosis plays an active role in the epithelial invagination of the Drosophila melanogaster tracheal placode. Invagination begins with a slow phase under the control of epidermal growth factor receptor (EGFR) signalling; in this process, the central apically constricted cells, which are surrounded by intercalating cells6,7, form a shallow pit. This slow phase is followed by a fast phase, in which the pit is rapidly depressed, accompanied by mitotic entry, which leads to the internalization of all the cells in the placode. We found that mitotic cell rounding, but not cell division, of the central cells in the placode is required to accelerate invagination, in conjunction with EGFR-induced myosin II contractility in the surrounding cells. We propose that mitotic cell rounding causes the epithelium to buckle under pressure and acts as a switch for morphogenetic transition at the appropriate time.

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Figure 1: Two-step process of tracheal invagination.
Figure 2: Mitosis is required for the acceleration of invagination.
Figure 3: Mitotic cell rounding is sufficient for the fast transition.
Figure 4: Mitosis triggers invagination independent of EGF and FGF signalling.


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We thank the Kyoto and Bloomington Drosophila Stock Centers, the Developmental Studies Hybridoma Bank, E. Wieschaus, H. Oda and T. Nishimura for fly stocks and antibodies; K. Kato and H. Wada for assistance with data analysis and experiments; G. Sheng, E. Kuranaga, K. Kato, T. Otani and B. Dong for comments on the manuscript; and members of the Hayashi, Nishimura and Kuranaga laboratories for discussions. This work was supported by Grant-in-Aid for Scientific Research on Innovative Areas (22111007 to S.H.); Grant-in-Aid for Young Scientists (B) (23770624 to T.K.) from The Ministry of Education, Culture, Sports, Science and Technology, Japan; and the RIKEN Special Postdoctoral Researcher Program (T.K.)

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Authors and Affiliations



T.K. and S.H. conceived the project and wrote the manuscript, and T.K. performed the experiments.

Corresponding author

Correspondence to Shigeo Hayashi.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-11, Supplementary Notes and additional references. (PDF 10689 kb)

Invagination of the tracheal placode

An embryo expressing E-cad-GFP (green) and His-RFP (magenta) Scale bar, 10 μm. (Related to Fig. 1a-c). (MOV 3698 kb)

Mitosis in the tracheal pit

An embryo expressing membrane-GFP. Scale bar, 10 μm. (Related to Fig. 1d). (MOV 2471 kb)

Invagination in a Cyclin A mutant embryo

A CycA mutant embryo expressing E-cad-GFP (green) and His-RFP (magenta). Scale bar, 10 μm. (Related to Fig. 2b). (MOV 2356 kb)

Invagination in a CycA bnl double mutant embryo

A CycA bnl double-mutant embryo expressing E-cad-GFP (green) and His-RFP (magenta) Scale bar, 10 μm. (Related to Fig. 2c). (MOV 2169 kb)

Invagination in a colchicine-treated embryo

A colchicine-treated embryo expressing E-cad-GFP (green) and His-RFP (magenta) Scale bar, 10 μm. (Related to Fig. 3). (MOV 3947 kb)

Invagination in a rho bnl double-mutant embryo

A rho bnl double-mutant embryo expressing Par-6-GFP (green) and His-RFP (magenta) Scale bar, 10 μm. (Related to Fig. 4e-h). (MOV 4456 kb)

Laser ablation of a non-tracheal cell

Laser ablation at the medial region of the apical surface of a surrounding cell. E-cad-GFP (green) and Myosin-mCherry (magenta). xy-view is the projection view of z2-6. Scale bar, 10 μm. (Related to Supplementary Fig. 9d-g). (MOV 524 kb)

Laser ablation of a tracheal cell at the apical-surface level

Laser ablation at the medial region of a central cell at the apical-surface level. E-cad-GFP (green) and Myosin-mCherry (magenta). xy-view is the projection view of z3-6. Scale bar, 10 μm. (Related to Supplementary Fig. 9h-j). (MOV 457 kb)

Laser ablation of a tracheal cell at the adherens junction level

Laser ablation at the medial region of a central cell at the adherens junction level. E-cad-GFP (green) and Myosin-mCherry (magenta). xy-view is the projection view of z3-6. Scale bar, 10 μm. (Related to Supplementary Fig. 9k-m). (MOV 474 kb)

Invagination in a rho CycA bnl triple mutant embryo

A rho CycA bnl triple-mutant embryo expressing Par-6-GFP (green) and His-RFP (magenta) Scale bar, 10 μm. (Related to Fig. 4i-l). (MOV 2506 kb)

Invagination in a rho CycA double mutant embryo

A rho CycA double-mutant embryo expressing Par-6-GFP (green) and His-RFP (magenta) Scale bar, 10 μm. (Related to Fig. S10i-l). (MOV 2286 kb)

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Kondo, T., Hayashi, S. Mitotic cell rounding accelerates epithelial invagination. Nature 494, 125–129 (2013).

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