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Prolonged fasting followed by refeeding modifies proteome profile and parvalbumin expression in the fast-twitch muscle of pacu (Piaractus mesopotamicus)


Autoři: Rafaela Nunes da Silva-Gomes aff001;  Maria Laura Gabriel Kuniyoshi aff001;  Bruno Oliveira da Silva Duran aff001;  Bruna Tereza Thomazini Zanella aff001;  Paula Paccielli Freire aff001;  Tassiana Gutierrez de Paula aff001;  Bruno Evaristo de Almeida Fantinatti aff001;  Rondinelle Artur Simões Salomão aff002;  Robson Francisco Carvalho aff001;  Lucilene Delazari Santos aff003;  Maeli Dal-Pai-Silva aff001
Působiště autorů: Department of Morphology, Institute of Bioscience of Botucatu, São Paulo State University (UNESP), Botucatu, São Paulo, Brazil aff001;  University of Western São Paulo (UNOESTE), Presidente Prudente, São Paulo, Brazil aff002;  Center for the Studies of Venoms and Venomous Animals (CEVAP)/ Graduate Program in Tropical Diseases (FMB), São Paulo State University (UNESP), Botucatu, São Paulo, Brazil aff003
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0225864

Souhrn

Here, we analyzed the fast-twitch muscle of juvenile Piaractus mesopotamicus (pacu) submitted to prolonged fasting (30d) and refeeding (6h, 24h, 48h and 30d). We measured the relative rate of weight and length increase (RRIlength and RRIweight), performed shotgun proteomic analysis and did Western blotting for PVALB after 30d of fasting and 30d of refeeding. We assessed the gene expression of igf-1, mafbx and pvalb after 30d of fasting and after 6h, 24h, 48h and 30d of refeeding. We performed a bioinformatic analysis to predict miRNAs that possibly control parvalbumin expression. After fasting, RRIlength, RRIweight and igf-1 expression decreased, while the mafbx expression increased, which suggest that prolonged fasting caused muscle atrophy. After 6h and 24h of refeeding, mafbx was not changed and igf-1 was downregulated, while after 48h of refeeding mafbx was downregulated and igf-1 was not changed. After 30d of refeeding, RRIlength and RRIweight were increased and igf-1 and mafbx expression were not changed. Proteomic analysis identified 99 proteins after 30d of fasting and 71 proteins after 30d of refeeding, of which 23 and 17, respectively, were differentially expressed. Most of these differentially expressed proteins were related to cytoskeleton, muscle contraction, and metabolism. Among these, parvalbumin (PVALB) was selected for further validation. The analysis showed that pvalb mRNA was downregulated after 6h and 24h of refeeding, but was not changed after 30d of fasting or 48h and 30d of refeeding. The Western blotting confirmed that PVALB protein was downregulated after 30d of fasting and 30d of refeeding. The downregulation of the protein and the unchanged expression of the mRNA after 30d of fasting and 30d of refeeding suggest a post-transcriptional regulation of PVALB. Our miRNA analysis predicted 444 unique miRNAs that may target pvalb. In conclusion, muscle atrophy and partial compensatory growth caused by prolonged fasting followed by refeeding affected the muscle proteome and PVALB expression.

Klíčová slova:

3' UTR – Gene expression – MicroRNAs – Muscle proteins – Protein expression – Protein interaction networks – Proteomics – Skeletal muscles


Zdroje

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