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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.
J Exp Biol. 2020 Jan 29; 223(Pt 2)JE

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-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 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.

Authors+Show Affiliations

São Paulo State University (UNESP), Institute of Biosciences, Department of Morphology, Botucatu 18618-689, São Paulo, Brazil.São Paulo State University (UNESP), Institute of Biosciences, Department of Morphology, Botucatu 18618-689, São Paulo, Brazil maeli.dal-pai@unesp.br.University of St Andrews, Scottish Oceans Institute, School of Biology, St Andrews, Fife KY16 8LB, UK. University of Barcelona, Faculty of Biology, Department of Cell Biology, Physiology and Immunology, 08028 Barcelona, Spain.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31871118

Citation

Duran, Bruno Oliveira da Silva, et al. "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." The Journal of Experimental Biology, vol. 223, no. Pt 2, 2020.
Duran BODS, Dal-Pai-Silva M, Garcia de la Serrana D. 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. J Exp Biol. 2020;223(Pt 2).
Duran, B. O. D. S., Dal-Pai-Silva, M., & Garcia de la Serrana, D. (2020). 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. The Journal of Experimental Biology, 223(Pt 2). https://doi.org/10.1242/jeb.216390
Duran BODS, Dal-Pai-Silva M, Garcia de la Serrana D. 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. J Exp Biol. 2020 Jan 29;223(Pt 2) PubMed PMID: 31871118.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Duran,Bruno Oliveira da Silva, AU - Dal-Pai-Silva,Maeli, AU - Garcia de la Serrana,Daniel, Y1 - 2020/01/29/ PY - 2019/10/14/received PY - 2019/12/17/accepted PY - 2019/12/25/pubmed PY - 2019/12/25/medline PY - 2019/12/25/entrez KW - Cell culture, Myoblasts KW - Slow skeletal muscle KW - miRNA, Electrostimulation JF - The Journal of experimental biology JO - J. Exp. Biol. VL - 223 IS - Pt 2 N2 - 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 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. SN - 1477-9145 UR - https://www.unboundmedicine.com/medline/citation/31871118/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 L2 - http://jeb.biologists.org/cgi/pmidlookup?view=long&pmid=31871118 DB - PRIME DP - Unbound Medicine ER -
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