Amino Acids and IGF1 Regulation of Fish Muscle Growth Revealed by Transcriptome and microRNAome Integrative Analyses of Pacu (Piaractus mesopotamicus) Myotubes.

Bruno Oliveira Silva Duran,Josefina Blasco,Daniel Garcia De La Serrana,Bruna Tereza Thomazini Zanella,Maeli Dal-Pai-Silva,Erika Stefani Perez,Edson Assunção Mareco

doi:10.3390/ijms23031180

Abstract

Amino acids (AA) and IGF1 have been demonstrated to play essential roles in protein synthesis and fish muscle growth. The myoblast cell culture is useful for studying muscle regulation, and omics data have contributed enormously to understanding its molecular biology. However, to our knowledge, no study has performed the large-scale sequencing of fish-cultured muscle cells stimulated with pro-growth signals. In this work, we obtained the transcriptome and microRNAome of pacu (Piaractus mesopotamicus)-cultured myotubes treated with AA or IGF1. We identified 1228 and 534 genes differentially expressed by AA and IGF1. An enrichment analysis showed that AA treatment induced chromosomal changes, mitosis, and muscle differentiation, while IGF1 modulated IGF/PI3K signaling, metabolic alteration, and matrix structure. In addition, potential molecular markers were similarly modulated by both treatments. Muscle-miRNAs (miR-1, -133, -206 and -499) were up-regulated, especially in AA samples, and we identified molecular networks with omics integration. Two pairs of genes and miRNAs demonstrated a high-level relationship, and involvement in myogenesis and muscle growth: marcksb and miR-29b in AA, and mmp14b and miR-338-5p in IGF1. Our work helps to elucidate fish muscle physiology and metabolism, highlights potential molecular markers, and creates a perspective for improvements in aquaculture and in in vitro meat production.

Highlights

The skeletal muscle in teleost fish represents up to 60% of its total body mass and is the most abundant tissue, with a set of characteristics necessary for fish physiology and metabolism, and with great importance for aquaculture industry [1,2]
Our work showed significant negative correlation between miRNAs and validated targets involved with myogenesis and muscle growth (Supplementary Figure S5), and the high enrichment of myogenic processes in Amino acids (AA) myotubes could be explained by the up-regulation of several muscle-specific miRNAs
We obtained transcriptomic and microRNAomic data from pacu-cultured myotubes submitted to AA and IGF1 treatments

Summary

Introduction

The skeletal muscle in teleost fish represents up to 60% of its total body mass and is the most abundant tissue, with a set of characteristics necessary for fish physiology and metabolism, and with great importance for aquaculture industry [1,2]. Muscle growth is a multifactorial process regulated by extrinsic and intrinsic signals. Extrinsic factors include nutrient availability, temperature, salinity, oxygenation, photoperiod, pH, and water flow [3]. Intrinsic signals include transcription factors (such as myogenic regulatory factors, MRFs), hormones, cytokines, and growth factors. These inputs can shift the balance between protein synthesis and degradation pathways, promoting protein accretion by favoring protein synthesis, and muscle growth [4]. Protein synthesis is strongly regulated by the IGF/PI3K/MTOR axis, while protein degradation is mediated by the ubiquitin-proteasome, calpain/calpastatin and autophagic-lysosomal systems [5,6]

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Results

Discussion

Conclusion