
pmid: 25893916
The closure of epidermal openings is an essential biological process that causes major developmental problems such as spina bifida in humans if it goes awry. At present, the mechanism of closure remains elusive. Therefore, we reconstructed a model closure event, dorsal closure in fly embryos, by large-volume correlative electron tomography. We present a comprehensive, quantitative analysis of the cytoskeletal reorganization, enabling separated epidermal cells to seal the epithelium. After establishing contact through actin-driven exploratory filopodia, cells use a single lamella to generate 'roof tile'-like overlaps. These shorten to produce the force, 'zipping' the tissue closed. The shortening overlaps lack detectable actin filament ensembles but are crowded with microtubules. Cortical accumulation of shrinking microtubule ends suggests a force generation mechanism in which cortical motors pull on microtubule ends as for mitotic spindle positioning. In addition, microtubules orient filopodia and lamellae before zipping. Our 4D electron microscopy picture describes an entire developmental process and provides fundamental insight into epidermal closure.
Electron Microscope Tomography, Microscopy, Confocal, Microscopy, Video, Microtubules, 10124 Institute of Molecular Life Sciences, Actins, Epithelium, 1307 Cell Biology, Animals, Genetically Modified, Luminescent Proteins, Drosophila melanogaster, Imaging, Three-Dimensional, Microscopy, Fluorescence, Genes, Reporter, Image Processing, Computer-Assisted, 570 Life sciences; biology, Animals, Pseudopodia, Cytoskeleton
Electron Microscope Tomography, Microscopy, Confocal, Microscopy, Video, Microtubules, 10124 Institute of Molecular Life Sciences, Actins, Epithelium, 1307 Cell Biology, Animals, Genetically Modified, Luminescent Proteins, Drosophila melanogaster, Imaging, Three-Dimensional, Microscopy, Fluorescence, Genes, Reporter, Image Processing, Computer-Assisted, 570 Life sciences; biology, Animals, Pseudopodia, Cytoskeleton
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