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Coupling optogenetics and light-sheet microscopy, a method to study Wnt signaling during embryogenesis
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Kaur, Prameet, Saunders, Timothy E. and Tolwinski, Nicholas S. (2017) Coupling optogenetics and light-sheet microscopy, a method to study Wnt signaling during embryogenesis. Scientific Reports, 7 (1). 16636. doi:10.1038/s41598-017-16879-0
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Official URL: http://dx.doi.org/10.1038/s41598-017-16879-0
Abstract
Optogenetics allows precise, fast and reversible intervention in biological processes. Light-sheet microscopy allows observation of the full course of Drosophila embryonic development from egg to larva. Bringing the two approaches together allows unparalleled precision into the temporal regulation of signaling pathways and cellular processes in vivo. To develop this method, we investigated the regulation of canonical Wnt signaling during anterior-posterior patterning of the Drosophila embryonic epidermis. Cryptochrome 2 (CRY2) from Arabidopsis Thaliana was fused to mCherry fluorescent protein and Drosophila β–catenin to form an easy to visualize optogenetic switch. Blue light illumination caused oligomerization of the fusion protein and inhibited downstream Wnt signaling in vitro and in vivo. Temporal inactivation of β–catenin confirmed that Wnt signaling is required not only for Drosophila pattern formation, but also for maintenance later in development. We anticipate that this method will be easily extendable to other developmental signaling pathways and many other experimental systems.
| Item Type: | Journal Article | ||||||||||||
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| Subjects: | Q Science > QH Natural history Q Science > QP Physiology T Technology > TA Engineering (General). Civil engineering (General) |
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| Divisions: | Faculty of Medicine > Warwick Medical School > Biomedical Sciences Faculty of Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | Optogenetics, Microscopy , Wnt pathway , Cellular signal transduction , Embryology | ||||||||||||
| Journal or Publication Title: | Scientific Reports | ||||||||||||
| Publisher: | Nature Publishing Group | ||||||||||||
| ISSN: | 2045-2322 | ||||||||||||
| Official Date: | 30 November 2017 | ||||||||||||
| Dates: |
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| Volume: | 7 | ||||||||||||
| Number: | 1 | ||||||||||||
| Article Number: | 16636 | ||||||||||||
| DOI: | 10.1038/s41598-017-16879-0 | ||||||||||||
| Status: | Peer Reviewed | ||||||||||||
| Publication Status: | Published | ||||||||||||
| Access rights to Published version: | Open Access | ||||||||||||
| RIOXX Funder/Project Grant: |
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