ZFIN ID: ZDB-PUB-200225-41
Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain
Shahar, O.D., Schuman, E.M.
Date: 2020
Source: eLIFE   9: (Journal)
Registered Authors: Schuman, Erin
Keywords: cell biology, cell type specific, neuron specific, neuroscience, protein synthesis, translation, translation control, zebrafish
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Brain/metabolism*
  • Nerve Tissue Proteins/biosynthesis*
  • Zebrafish
  • Zebrafish Proteins/biosynthesis*
PubMed: 32091983 Full text @ Elife
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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.
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