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

• Published in: Journal of Experimental Biology
• Main Authors: da Silva Duran, Bruno Oliveira, Dal-Pai-Silva, Maeli, de la Serrana, Daniel Garcia
• Other Authors: Universidade Estadual Paulista (UNESP)
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2020
• Subjects: Cell culture; Electrostimulation; MiRNA; Myoblasts; Slow skeletal muscle; Atlantic salmon
• Online Access: http://hdl.handle.net/11449/198465; https://doi.org/10.1242/jeb.216390

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Scottish Funding Council Processo FAPESP: 2015/03234-8 Processo FAPESP: 2016/05009-4 Processo FAPESP: 2016/19683-9 Processo FAPESP: 2019/01592-5 CNPq: 302656/2015-4 CNPq: 447233/2014 Scottish Funding Council: HR09011 Muscle 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 ...