Optogenetic control of Nodal signaling patterns.

Autor:	McNamara HM; Lewis Sigler Institute, Princeton University, Princeton, NJ, USA., Jia BZ; Department of Systems Biology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA.; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA., Guyer A; Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA., Parot VJ; Institute for Biological and Medical Engineering, Pontificia Universidad Católica de Chile, Santiago, Chile., Dobbs C; Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA., Schier AF; Biozentrum, University of Basel, Basel, Switzerland., Cohen AE; Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA, USA.; Department of Physics, Harvard University, Cambridge, MA, USA., Lord ND; Department of Computational and Systems Biology, University of Pittsburgh, Pittsburgh, PA, USA.; McGowan Institute of Regenerative Medicine, University of Pittsburgh, Pittsburgh, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Apr 12. Date of Electronic Publication: 2024 Apr 12.
DOI:	10.1101/2024.04.11.588875
Abstrakt:	A crucial step in early embryogenesis is the establishment of spatial patterns of signaling activity. Tools to perturb morphogen signals with high resolution in space and time can help reveal how embryonic cells decode these signals to make appropriate fate decisions. Here, we present new optogenetic reagents and an experimental pipeline for creaHng designer Nodal signaling patterns in live zebrafish embryos. Nodal receptors were fused to the light-sensitive heterodimerizing pair Cry2/CIB1N, and the Type II receptor was sequestered to the cytosol. The improved optoNodal2 reagents eliminate dark activity and improve response kinetics, without sacrificing dynamic range. We adapted an ultra-widefield microscopy platform for parallel light patterning in up to 36 embryos and demonstrated precise spatial control over Nodal signaling activity and downstream gene expression. Patterned Nodal activation drove precisely controlled internalization of endodermal precursors. Further, we used patterned illumination to generate synthetic signaling patterns in Nodal signaling mutants, rescuing several characteristic developmental defects. This study establishes an experimental toolkit for systematic exploration of Nodal signaling patterns in live embryos.
Databáze:	MEDLINE
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