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In vivo photocontrol of microtubule dynamics and integrity, migration and mitosis, by the potent GFP-imaging-compatible photoswitchable reagents SBTubA4P and SBTub2M

Authors :
Anna Akhmanova
Constanze Heise
M. Wranik
Oliver Thorn-Seshold
Clemens Cabernard
Li Gao
Adam Varady
Jörg Standfuss
Jennifer A. Taylor
Joyce C.M. Meiring
Andreas R. Bausch
Beatrice Terni
Iris E. Ruider
Artur Llobet
Martin Distel
Cecilia D. Velasco
Michel O. Steinmetz
Julia Thorn-Seshold
Publication Year :
2021
Publisher :
Cold Spring Harbor Laboratory, 2021.

Abstract

Photoswitchable reagents to modulate microtubule stability and dynamics are an exciting tool approach towards micron- and millisecond-scale control over endogenous cytoskeleton-dependent processes. When these reagents are globally administered yet locally photoactivated in 2D cell culture, they can exert precise biological control that would have great potential forin vivotranslation across a variety of research fields and for all eukaryotes. However, photopharmacology’s reliance on the azobenzene photoswitch scaffold has been accompanied by a failure to translate this temporally- and cellularly-resolved control to 3D models or toin vivoapplications in multi-organ animals, which we attribute substantially to the metabolic liabilities of azobenzenes.Here, we optimised the potency and solubility of metabolically stable, druglike colchicinoid microtubule inhibitors based instead on the styrylbenzothiazole (SBT) photoswitch scaffold, that are non-responsive to the major fluorescent protein imaging channels and so enable multiplexed imaging studies. We applied these reagents to 3D systems (organoids, tissue explants) and classic model organisms (zebrafish, clawed frog) with one- and two-protein imaging experiments. We successfully used systemic treatment plus spatiotemporally-localised illuminationsin vivoto photocontrol microtubule dynamics, network architecture, and microtubule-dependent processes in these systems with cellular precision and second-level resolution. These nanomolar,in vivo-capable photoswitchable reagents can prove a game-changer for high-precision cytoskeleton research in cargo transport, cell motility, cell division and development. More broadly, their straightforward design can also inspire the development of similarly capable optical reagents for a range of protein targets, so bringing generalin vivophotopharmacology one step closer to productive realisation.

Details

Database :
OpenAIRE
Accession number :
edsair.doi...........764649c00e9b219494130e1c036d2e93
Full Text :
https://doi.org/10.1101/2021.03.26.437160