Coupling optogenetics and light-sheet microscopy to study signal transduction in vivo
作者： Prameet Kaur , Timothy E. Saunders , Nicholas S. Tolwinski
DOI: 10.1101/114686
关键词: Signal transduction 、 Optogenetics 、 Arabidopsis thaliana 、 Biology 、 Fusion protein 、 Wnt signaling pathway 、 Developmental biology 、 Cryptochrome 、 Cell biology 、 Light sheet fluorescence microscopy
摘要: Optogenetics allows precise, fast and reversible intervention in biological processes. Coupled with light-sheet microscopy, this approach unparalleled insight into the regulation of signaling pathways cellular processes from a spatial temporal perspective vivo. To develop method, we investigated canonical Wnt during anterior-posterior patterning Drosophila embryonic epidermis. Cryptochrome C (Cry2) Arabidopsis Thaliana was fused to mCherry fluorescent protein β-catenin form an easy visualize optogenetic switch. Blue light illumination caused oligomerization fusion inhibited downstream vitro Temporal inactivation showed that is required not only for formation, but also maintenance throughout development. By applying microscopy activate pathway then record subsequent behavior toto, show precise We anticipate method will be easily extendable other developmental many experimental systems.
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