Rainbow trout slow myoblast cell culture as a model to study slow skeletal muscle, and the characterization of mir-133 and mir-499 families as a case study

dc.contributor.authorda Silva Duran, Bruno Oliveira [UNESP]
dc.contributor.authorDal-Pai-Silva, Maeli [UNESP]
dc.contributor.authorde la Serrana, Daniel Garcia
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionSchool of Biology
dc.contributor.institutionPhysiology and Immunology
dc.description.abstractMuscle fibres are classified as fast, intermediate and slow. In vitro myoblast cell culture model from fast muscle is a very useful tool to study muscle growth and development; however, similar models for slow muscle do not exist. Owing to the compartmentalization of fish muscle fibres, we have developed a slow myoblast cell culture for rainbow trout (Oncorhynchus mykiss). Slow and fast muscle-derived myoblasts have similar morphology, but with differential expression of slow muscle markers such as slow myhc, sox6 and pgc-1α. We also characterized the mir-133 and mir-499 microRNA families in trout slow and fast myoblasts as a case study during myogenesis and in response to electrostimulation. Three mir-133 (a-1a, a-1b and a-2) and four mir-499 (aa, ab, ba and bb) paralogues were identified for rainbow trout and named base on their phylogenetic relationship to zebrafish and Atlantic salmon orthologues. Omy-mir-499ab and omymir-499bb had 0.6 and 0.5-fold higher expression in slow myoblasts compared with fast myoblasts, whereas mir-133 duplicates had similar levels in both phenotypes and little variation during development. Slow myoblasts also showed increased expression for omy-mir-499b paralogues in response to chronic electrostimulation (7-fold increase for omy-mir-499ba and 2.5-fold increase for omy-mir-499bb). The higher expression of mir-499 paralogues in slow myoblasts suggests a role in phenotype determination, while the lack of significant differences of mir-133 copies during culture development might indicate a different role in fish compared with mammals. We have also found signs of sub-functionalization of mir-499 paralogues after electrostimulation, with omy-mir-499b copies more responsive to electrical signals.en
dc.description.affiliationSaõ Paulo State University (UNESP) Institute of Biosciences Department of Morphology
dc.description.affiliationUniversity of St.Andrews Scottish Oceans Institute School of Biology
dc.description.affiliationUniversity of Barcelona Faculty of Biology Department of Cell Biology Physiology and Immunology
dc.description.affiliationUnespSaõ Paulo State University (UNESP) Institute of Biosciences Department of Morphology
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipScottish Funding Council
dc.description.sponsorshipIdFAPESP: 2015/03234-8
dc.description.sponsorshipIdFAPESP: 2016/05009-4
dc.description.sponsorshipIdFAPESP: 2016/19683-9
dc.description.sponsorshipIdFAPESP: 2019/01592-5
dc.description.sponsorshipIdCNPq: 302656/2015-4
dc.description.sponsorshipIdCNPq: 447233/2014
dc.description.sponsorshipIdScottish Funding Council: HR09011
dc.identifier.citationJournal of Experimental Biology, v. 223, n. 2, 2020.
dc.relation.ispartofJournal of Experimental Biology
dc.subjectCell culture
dc.subjectSlow skeletal muscle
dc.titleRainbow trout slow myoblast cell culture as a model to study slow skeletal muscle, and the characterization of mir-133 and mir-499 families as a case studyen