Multi-view light-sheet imaging and tracking with the MaMuT software reveals the cell lineage of a direct developing arthropod limb
- Humboldt- Universität zu Berlin, Germany
- Institut Pasteur, France
- Max Planck Institute of Molecular Cell Biology and Genetics, Germany
- Janelia Farm Research Campus, Howard Hughes Medical Institute, United States
- Max Delbrück Center for Molecular Medicine, Germany
- Max Planck Institute of Cell Biology and Genetics, Germany
Abstract
During development, coordinated cell behaviors orchestrate tissue and organ morphogenesis. Detailed descriptions of cell lineages and behaviors provide a powerful framework to elucidate the mechanisms of morphogenesis. To study the cellular basis of limb development, we imaged transgenic fluorescently-labeled embryos from the crustacean Parhyale hawaiensis with multi-view light-sheet microscopy at high spatiotemporal resolution over several days of embryogenesis. The cell lineage of outgrowing thoracic limbs was reconstructed at single-cell resolution with new software called Massive Multi-view Tracker (MaMuT). In silico clonal analyses suggested that the early limb primordium becomes subdivided into anterior-posterior and dorsal-ventral compartments whose boundaries intersect at the distal tip of the growing limb. Limb-bud formation is associated with spatial modulation of cell proliferation, while limb elongation is also driven by preferential orientation of cell divisions along the proximal-distal growth axis. Cellular reconstructions were predictive of the expression patterns of limb development genes including the BMP morphogen Decapentaplegic.
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Author details
Anastasios Pavlopoulos
Funding
Howard Hughes Medical Institute
- Evangelia Stamataki
- Léo Guignard
- Philipp J Keller
- Anastasios Pavlopoulos
European Commission Marie Sklodowska-Curie Actions (FP7-IEF 302235)
- Anastasios Pavlopoulos
Max-Planck-Institute of Molecular Cell Biology and Genetics
- Tobias Pietzsch
- Benjamin Harich
- Pavel Tomancak
European Research Council (260746)
- Tobias Pietzsch
- Pavel Tomancak
Einstein Stiftung Berlin (A-2012_114)
- Carsten Wolff
Institut Pasteur
- Jean-Yves Tinevez
- Spencer Shorte
Agence Nationale de la Recherche
- Jean-Yves Tinevez
- Spencer Shorte
Helmholtz-Gemeinschaft
- Stephan Preibisch
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Copyright
© 2018, Wolff et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Multi-view light-sheet imaging and tracking with the MaMuT software reveals the cell lineage of a direct developing arthropod limb
eLife 7:e34410.
https://doi.org/10.7554/eLife.34410
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