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Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain.

Authors :
Shahar OD
Schuman EM
Source :
ELife [Elife] 2020 Feb 24; Vol. 9. Date of Electronic Publication: 2020 Feb 24.
Publication Year :
2020

Abstract

Despite advances in methods to detect protein synthesis, it has not been possible to measure endogenous protein synthesis levels in vivo in an entire vertebrate brain. We developed a transgenic zebrafish line that allows for cell-type-specific labeling and imaging of nascent proteins in the entire animal. By replacing leucine with glycine in the zebrafish MetRS-binding pocket (MetRS-L270G), we enabled the cell-type-specific incorporation of the azide-bearing non-canonical-amino-acid azidonorleucine (ANL) during protein synthesis. Newly synthesized proteins were then labeled via 'click chemistry'. Using a Gal4-UAS-ELAV3 line to express MetRS-L270G in neurons, we measured protein synthesis intensities across the entire nervous system. We visualized endogenous protein synthesis and demonstrated that seizure-induced neural activity results in enhanced translation levels in neurons. This method allows for robust analysis of endogenous protein synthesis in a cell-type-specific manner, in vivo at single-cell resolution.<br />Competing Interests: OS, ES No competing interests declared<br /> (© 2020, Shahar and Schuman.)

Details

Language :
English
ISSN :
2050-084X
Volume :
9
Database :
MEDLINE
Journal :
ELife
Publication Type :
Academic Journal
Accession number :
32091983
Full Text :
https://doi.org/10.7554/eLife.50564