PUBLICATION
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
- Authors
- Duran, B.O.D.S., Dal-Pai-Silva, M., Garcia de la Serrana, D.
- ID
- ZDB-PUB-191226-21
- Date
- 2019
- Source
- The Journal of experimental biology 223(Pt 2): (Journal)
- Registered Authors
- Keywords
- Cell culture, Electrostimulation, Myoblasts, Slow skeletal muscle, miRNA
- MeSH Terms
-
- Animals
- Cell Culture Techniques/methods
- MicroRNAs/metabolism*
- Muscle Development
- Muscle Fibers, Fast-Twitch/metabolism
- Muscle Fibers, Slow-Twitch/metabolism
- Myoblasts, Skeletal/metabolism
- Myoblasts, Skeletal/physiology*
- Oncorhynchus mykiss*
- PubMed
- 31871118 Full text @ J. Exp. Biol.
Citation
Duran, B.O.D.S., Dal-Pai-Silva, M., Garcia de la Serrana, D. (2019) 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):.
Abstract
Muscle fibers 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. Thanks to the compartmentalization of fish muscle fibers we developed a slow myoblast cell culture for rainbow trout (Oncorhynchus mykiss). Slow and fast muscle-derived myoblasts had 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 to 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 to 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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping