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Live Imaging of RNA Transport and Translation in Xenopus Retinal Axons.

Live Imaging of RNA Transport and Translation in Xenopus Retinal Axons.

Authors :
Lin JQ
Cioni JM
Source :
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2431, pp. 49-69.
Publication Year :
2022

Abstract

In neurons, specific mRNAs are transported into axons, where their local translation supports essential cellular functions. Over the years, our knowledge of the molecular mechanisms underlying axonal mRNA translation has rapidly expanded. However, tools to study mRNA localization and translation in real time with high spatial precision were not available until recently. Here, we present a live imaging approach to examine axonal mRNA trafficking and translation simultaneously in Xenopus retinal ganglion cells (RGCs), using in vitro synthesized fluorescently labeled mRNAs coupled with a genetically encoded protein tagging system to visualize synthesizing peptides at single-molecule resolution. We further describe the process of image analysis in detail, thus providing a methodology that can be used to investigate new research questions in the field.<br /> (© 2022. The Author(s).)

Details

Language :
English
ISSN :
1940-6029
Volume :
2431
Database :
MEDLINE
Journal :
Methods in molecular biology (Clifton, N.J.)
Publication Type :
Academic Journal
Accession number :
35412271
Full Text :
https://doi.org/10.1007/978-1-0716-1990-2_3