Axon microdissection and transcriptome profiling reveals the in vivo RNA content of fully differentiated myelinated motor axons.

Autor:	Farias J; Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, 11600, Uruguay.; Departamento de Proteínas y Ácidos Nucléicos, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, 11600, Uruguay., Holt CE; Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, CB2 3DY, United Kingdom., Sotelo JR; Departamento de Proteínas y Ácidos Nucléicos, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, 11600, Uruguay., Sotelo-Silveira JR; Departamento de Genómica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, 11600, Uruguay.; Departamento de Biología Celular y Molecular, Facultad de Ciencias, Universidad de la República, Montevideo, 11400, Uruguay.
Jazyk:	angličtina
Zdroj:	RNA (New York, N.Y.) [RNA] 2020 May; Vol. 26 (5), pp. 595-612. Date of Electronic Publication: 2020 Feb 12.
DOI:	10.1261/rna.073700.119
Abstrakt:	Axonal protein synthesis has been shown to play a role in developmental and regenerative growth, as well as in the maintenance of the axoplasm in a steady state. Recent studies have begun to identify the mRNAs localized in axons, which could be translated locally under different conditions. Despite that by now hundreds or thousands of mRNAs have been shown to be localized into the axonal compartment of cultured neurons in vitro, knowledge of which mRNAs are localized in mature myelinated axons is quite limited. With the purpose of characterizing the transcriptome of mature myelinated motor axons of peripheral nervous systems, we modified the axon microdissection method devised by Koenig, enabling the isolation of the axoplasm RNA to perform RNA-seq analysis. The transcriptome analysis indicates that the number of RNAs detected in mature axons is lower in comparison with in vitro data, depleted of glial markers, and enriched in neuronal markers. The mature myelinated axons are enriched for mRNAs related to cytoskeleton, translation, and oxidative phosphorylation. Moreover, it was possible to define core genes present in axons when comparing our data with transcriptomic data of axons grown in different conditions. This work provides evidence that axon microdissection is a valuable method to obtain genome-wide data from mature and myelinated axons of the peripheral nervous system, and could be especially useful for the study of axonal involvement in neurodegenerative pathologies of motor neurons such as amyotrophic lateral sclerosis (ALS) and spinal muscular atrophies (SMA). (© 2020 Farias et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)
Databáze:	MEDLINE
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