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Tissue Mechanics Regulate Mitotic Nuclear Dynamics during Epithelial Development.

Current biology : CB (2020-05-16)
Natalie J Kirkland, Alice C Yuen, Melda Tozluoglu, Nancy Hui, Ewa K Paluch, Yanlan Mao
ABSTRACT

Cell divisions are essential for tissue growth. In pseudostratified epithelia, where nuclei are staggered across the tissue, each nucleus migrates apically before undergoing mitosis. Successful apical nuclear migration is critical for planar-orientated cell divisions in densely packed epithelia. Most previous investigations have focused on the local cellular mechanisms controlling nuclear migration. Inter-species and inter-organ comparisons of different pseudostratified epithelia suggest global tissue architecture may influence nuclear dynamics, but the underlying mechanisms remain elusive. Here, we use the developing Drosophila wing disc to systematically investigate, in a single epithelial type, how changes in tissue architecture during growth influence mitotic nuclear migration. We observe distinct nuclear dynamics at discrete developmental stages, as epithelial morphology changes. We use genetic and physical perturbations to show a direct effect of cell density on mitotic nuclear positioning. We find Rho kinase and Diaphanous, which facilitate mitotic cell rounding in confined cell conditions, are essential for efficient apical nuclear movement. Perturbation of Diaphanous causes increasing defects in apical nuclear migration as the tissue grows and cell density increases, and these defects can be reversed by acute physical reduction of cell density. Our findings reveal how the mechanical environment imposed on cells within a tissue alters the molecular and cellular mechanisms adopted by single cells for mitosis.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Shields and Sang M3 Insect Medium, With L-glutamine and potassium bicarbonate., liquid, sterile-filtered, suitable for insect cell culture
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
20-Hydroxyecdysone, ≥93% (HPLC), powder
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
4′,6-Diamidino-2-phenylindole dihydrochloride, powder, BioReagent, suitable for cell culture, ≥98% (HPLC and TLC), suitable for fluorescence