Coupling optogenetics and light-sheet microscopy, a method to study Wnt signaling during embryogenesis.

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.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Kaur P,Saunders TE,Tolwinski NS

doi

10.1038/s41598-017-16879-0

subject

Has Abstract

pub_date

2017-11-30 00:00:00

pages

16636

issue

1

issn

2045-2322

pii

10.1038/s41598-017-16879-0

journal_volume

7

pub_type

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