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

التفاصيل البيبلوغرافية
العنوان: Large-scale cell-type-specific imaging of protein synthesis in a vertebrate brain
المؤلفون: Or David Shahar, Erin Margaret Schuman
المصدر: eLife, Vol 9 (2020)
بيانات النشر: eLife Sciences Publications Ltd, 2020.
سنة النشر: 2020
المجموعة: LCC:Medicine
LCC:Science
LCC:Biology (General)
مصطلحات موضوعية: translation, cell type specific, zebrafish, protein synthesis, translation control, neuron specific, Medicine, Science, Biology (General), QH301-705.5
الوصف: 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.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2050-084X
Relation: https://elifesciences.org/articles/50564; https://doaj.org/toc/2050-084X
DOI: 10.7554/eLife.50564
URL الوصول: https://doaj.org/article/e962dd68dac842e2bfceaf3abd37965e
رقم الأكسشن: edsdoj.962dd68dac842e2bfceaf3abd37965e
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:2050084X
DOI:10.7554/eLife.50564