PUBLICATION
Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain
- Authors
- Shahar, O.D., Schuman, E.M.
- ID
- ZDB-PUB-200225-41
- 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
Citation
Shahar, O.D., Schuman, E.M. (2020) Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain. eLIFE. 9:.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping