Early Drosophila Spaghetti-Squash-GFP FRAP
Cite this dataset
Lefbvre, Matthew; Claussen, Nikolas (2023). Early Drosophila Spaghetti-Squash-GFP FRAP [Dataset]. Dryad. https://doi.org/10.25349/D94C8M
The actomyosin cytoskeleton is a crucial driver of morphogenesis. Yet how the behavior of large-scale cytoskeletal patterns in deforming tissues emerges from the interplay of geometry, genetics, and mechanics remains incompletely understood. Convergent extension flow in D. melanogaster embryos provides the opportunity to establish a quantitative understanding of the dynamics of anisotropic non-muscle myosin II. Cell-scale analysis of protein localization in fixed embryos suggests that there are complex rules governing how the control of myosin anisotropy is regulated by gene expression patterns. However, technical limitations have impeded quantitative and dynamic studies of this process at the whole embryo level, leaving the role of geometry open. Here we combine in toto live imaging with quantitative analysis of molecular dynamics to characterize the distribution of myosin anisotropy and corresponding genetic patterning. We found pair rule gene expression continuously deformed, flowing with the tissue frame. In contrast, myosin anisotropy orientation remained nearly static, aligned with the stationary dorsal-ventral axis of the embryo. We propose myosin recruitment by a geometrically defined static source, potentially related to the embryo-scale epithelial tension, and account for transient deflections by the interplay of cytoskeletal turnover with junction reorientation by flow. With only one parameter, this model quantitatively accounts for the time course of myosin anisotropy orientation in wild-type, twist, and even-skipped embryos as well as embryos with perturbed egg geometry. Geometric patterning of the cytoskeleton suggests a simple physical strategy to ensure a robust flow and formation of shape.
Fluorescence recovery after photobleaching (FRAP) of junctional myosin-II in the early Drosophila embryo. Data for the manuscript "Geometric control of Myosin-II orientation during axis elongation" by Lefebvre & Claussen et al.
Data consists of .tif stacks showing a full-time series of a FRAP experiment carried out on a Leica SP5 confocal microscope with a 63x/1.4 NA oil immersion objective at a frame rate of 1 frame per 1.78 seconds. Regions were bleached with 50mW laser power.
Software to view .tif files, e.g. Fiji.
National Institute of General Medical Sciences, Award: R35 GM138203