Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar

Abstract MicroRNAs are key non-coding RNA molecules that play a relevant role in the regulation of gene expression through translational repression and/or transcript cleavage during normal development and physiological adaptation processes like stress. Quantitative reverse transcription polymerase c...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Eduardo Zavala, Daniela Reyes, Robert Deerenberg, Rodrigo Vidal
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e35
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:a0d66163efbd49b69fcfcd67af706e35
record_format	dspace
spelling	oai:doaj.org-article:a0d66163efbd49b69fcfcd67af706e352021-12-02T16:06:42ZSelection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar10.1038/s41598-017-01970-32045-2322https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e352017-05-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-01970-3https://doaj.org/toc/2045-2322Abstract MicroRNAs are key non-coding RNA molecules that play a relevant role in the regulation of gene expression through translational repression and/or transcript cleavage during normal development and physiological adaptation processes like stress. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) has become the approach normally used to determine the levels of microRNAs. However, this approach needs the use of endogenous reference. An improper selection of endogenous references can result in confusing interpretation of data. The aim of this study was to identify and validate appropriate endogenous reference miRNA genes for normalizing RT-qPCR survey of miRNAs expression in four different tissues of Atlantic salmon, under handling and confinement stress conditions associated to early or primary stress response. Nine candidate reference normalizers, including microRNAs and nuclear genes, normally used in vertebrate microRNA expression studies were selected from literature, validated by RT-qPCR and analyzed by the algorithms geNorm and NormFinder. The results revealed that the ssa-miR-99-5p gene was the most stable overall and that ssa-miR-99-5p and ssa-miR-23a-5p genes were the best combination. Moreover, the suitability of ssa-miR-99-5p and ssa-miR-23a-5p as endogeneuos reference genes was demostrated by the expression analysis of ssa-miR-193-5p gene.Eduardo ZavalaDaniela ReyesRobert DeerenbergRodrigo VidalNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
description	Abstract MicroRNAs are key non-coding RNA molecules that play a relevant role in the regulation of gene expression through translational repression and/or transcript cleavage during normal development and physiological adaptation processes like stress. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) has become the approach normally used to determine the levels of microRNAs. However, this approach needs the use of endogenous reference. An improper selection of endogenous references can result in confusing interpretation of data. The aim of this study was to identify and validate appropriate endogenous reference miRNA genes for normalizing RT-qPCR survey of miRNAs expression in four different tissues of Atlantic salmon, under handling and confinement stress conditions associated to early or primary stress response. Nine candidate reference normalizers, including microRNAs and nuclear genes, normally used in vertebrate microRNA expression studies were selected from literature, validated by RT-qPCR and analyzed by the algorithms geNorm and NormFinder. The results revealed that the ssa-miR-99-5p gene was the most stable overall and that ssa-miR-99-5p and ssa-miR-23a-5p genes were the best combination. Moreover, the suitability of ssa-miR-99-5p and ssa-miR-23a-5p as endogeneuos reference genes was demostrated by the expression analysis of ssa-miR-193-5p gene.
format	article
author	Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal
author_facet	Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal
author_sort	Eduardo Zavala
title	Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
title_short	Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
title_full	Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
title_fullStr	Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
title_full_unstemmed	Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
title_sort	selection of reference genes for microrna analysis associated to early stress response to handling and confinement in salmo salar
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e35
work_keys_str_mv	AT eduardozavala selectionofreferencegenesformicrornaanalysisassociatedtoearlystressresponsetohandlingandconfinementinsalmosalar AT danielareyes selectionofreferencegenesformicrornaanalysisassociatedtoearlystressresponsetohandlingandconfinementinsalmosalar AT robertdeerenberg selectionofreferencegenesformicrornaanalysisassociatedtoearlystressresponsetohandlingandconfinementinsalmosalar AT rodrigovidal selectionofreferencegenesformicrornaanalysisassociatedtoearlystressresponsetohandlingandconfinementinsalmosalar
_version_	1718384920297996288

Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar

Ejemplares similares