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

Author(s)

da Silva Duran, Bruno Oliveira ^[1] ; Dal-Pai-Silva, Maeli ^{[1, 2]} ; Garcia de la Serrana, Daniel ^{[3, 4]}

Affiliation

^[1] Sao Paulo State Univ UNESP, Inst Biosci, Dept Morphol, BR-18618689 Botucatu, SP - Brazil

^[2] Garcia de la Serrana, Daniel, Univ Barcelona, Fac Biol, Dept Cell Biol Physiol \& Immunol, Barcelona 08028, Spain.da Silva Duran, Bruno Oliveira, Sao Paulo State Univ UNESP, Inst Biosci, Dept Morphol, BR-18618689 Botucatu, SP - Brazil

^[3] Univ St Andrews, Sch Biol, Scottish Oceans Inst, St Andrews KY16 8LB, Fife - Scotland

^[4] Univ Barcelona, Fac Biol, Dept Cell Biol Physiol & Immunol, Barcelona 08028 - Spain

• Document type: Journal article
• Source: Journal of Experimental Biology; v. 223, n. 2 JAN 2020.
• Web of Science Citations: 3

Abstract

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-1a. 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 omy-mir-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. (AU)

• FAPESP's process: 16/19683-9 - The slow myoblast cell culture as a model to understand the myogenesis and growth of slow-twitch muscle
• Grantee: Bruno Oliveira da Silva Duran
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• FAPESP's process: 15/03234-8 - The pacu (Piaractus mesopotamicus) myoblast cell culture as a model to understand muscle growth regulation in Ostariophysi superorder
• Grantee: Bruno Oliveira da Silva Duran
• Support Opportunities: Scholarships in Brazil - Doctorate
• FAPESP's process: 16/05009-4 - Comparative and integrative analysis of the transcriptome and the microRNAoma in skeletal muscle of Colossoma macropomum (tambaqui), Piaractus mesopotamicus (pacu) and its hybrid, tambacu: molecular, in silico, in vitro and in vivo approaches.
• Grantee: Maeli Dal Pai
• Support Opportunities: Regular Research Grants