Expressão de genes das vias anabólicas e catabólicas e de miRNas no músculo esquelético do Piaractus mesopotamicus durante período de jejum e realimentação
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2017-09-22
Autores
Paula, Tassiana Gutierrez De
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Universidade Estadual Paulista (Unesp)
Resumo
O músculo esquelético é capaz de adaptação fenotípica à fatores ambientais, como a disponibilidade de nutrientes, alterando o equilíbrio entre catabolismo e anabolismo muscular que, por sua vez, coordena o crescimento muscular. Os pequenos RNAs não codificados, conhecidos como microRNAs (miRNAs), reprimem a expressão de mRNAs alvo e muitos estudos demonstraram que os miRNAs regulam os mRNAs de genes catabólicos e anabólicos. Nós avaliamos a morfologia muscular, a expressão gênica de componentes envolvidos com o catabolismo, anabolismo e metabolismo energético e expressão de miRNAs no músculo rápido e lento de juvenis de pacus (Piaractus mesopotamicus) durante um período de restrição alimentar e realimentação. Nossa análise revelou que períodos curtos de restrição alimentar seguidos por realimentação afetaram predominantemente o músculo rápido, alterando o diâmetro da fibra muscular e a expressão de RNAs e miRNAs. Houve um aumento nos níveis de mRNA dos componentes catabólicos (FBXO25, ATG12, BCL2) e genes relacionados ao metabolismo energético (PGC1a e SDHA), juntamente com uma diminuição nos níveis de PPARβ / δmRNA. Curiosamente, um aumento nos níveis de mRNA dos genes anabólicos (PI3K e complexo mTORC1: mTOR, mLST8 e RAPTOR) também foi observado durante a restrição alimentar. Após realimentação, a morfologia muscular mostrou padrões similares do grupo controle; a maioria dos genes estavam ligeiramente regulados de forma up e down em mabas as musculaturas, respectivamente; os níveis de todos os miRNAs aumentaram no músculo rápido e alguns deles diminuíram em músculos lentos. Nossos achados demonstraram que um curto período de restrição alimentar no pacu juvenil teve um impacto considerável no músculo rápido, aumentando a expressão de genes anabolizantes (PI3K e mTORC1 complexos: mTOR, mLST8 e RAPTOR) e do metabolismo energético. Os miRNA (miR-1, miR-206, miR-199 e miR-23a) foram mais expressos durante a realimentação enquanto seus genes alvo (IGF-1, mTOR, PGC1α e MAFbx) apresentaram uma diminuição da expressão. As alterações no complexo mTORC1 observadas durante o jejum podem ter influenciado as taxas de síntese protéica usando aminoácidos da degradação protéica como mecanismo alternativo para preservar o fenótipo muscular e a manutenção da demanda metabólica.
Skeletal muscle is capable of phenotypic adaptation to environmental factors, such as nutrient availability, by altering the balance between muscle catabolism and anabolism that in turn coordinates muscle growth. Small noncoding RNAs, known as microRNAs (miRNAs), repress the expression of target mRNAs, and many studies have demonstrated that miRNAs regulate the mRNAs of catabolic and anabolic genes. We evaluated muscle morphology, gene expression of components involved in catabolism, anabolism and energetic metabolism and miRNAs expression in both the fast and slow muscle of juvenile pacu (Piaractus mesopotamicus) during food restriction and refeeding. Our analysis revealed that short periods of food restriction followed by refeeding predominantly affected fast muscle, with changes in muscle fiber diameter and miRNAs expression. There was an increase in the mRNA levels of catabolic pathways components (FBXO25, ATG12, BCL2) and energetic metabolism-related genes (PGC1αand SDHA), together with a decrease in PPARβ/δmRNA levels. Interestingly, an increase in mRNA levels of anabolic genes (PI3K and mTORC1 complex: mTOR, mLST8 and RAPTOR) was also observed during food restriction. After refeeding, muscle morphology showed similar patterns of the control group; the majority of genes were slightly up- or down-regulated in fast and slow muscle, respectively; the levels of all miRNAs increased in fast muscle and some of them decreased in slow muscle. Our findings demonstrated that a short period of food restriction in juvenile pacu had a considerable impact on fast muscle, increasing the expression of anabolic (PI3K and mTORC1 complex: mTOR, mLST8 and RAPTOR) and energetic metabolism genes. The miRNAs (miR-1, miR-206, miR-199 and miR-23a) were more expressed during refeeding and while their target genes (IGF-1, mTOR, PGC1α and MAFbx), presented a decreased expression. The alterations in mTORC1 complex observed during fasting may have influenced the rates of protein synthesis by using amino acids from protein degradation as an alternative mechanism to preserve muscle phenotype and metabolic demand maintenance.
Skeletal muscle is capable of phenotypic adaptation to environmental factors, such as nutrient availability, by altering the balance between muscle catabolism and anabolism that in turn coordinates muscle growth. Small noncoding RNAs, known as microRNAs (miRNAs), repress the expression of target mRNAs, and many studies have demonstrated that miRNAs regulate the mRNAs of catabolic and anabolic genes. We evaluated muscle morphology, gene expression of components involved in catabolism, anabolism and energetic metabolism and miRNAs expression in both the fast and slow muscle of juvenile pacu (Piaractus mesopotamicus) during food restriction and refeeding. Our analysis revealed that short periods of food restriction followed by refeeding predominantly affected fast muscle, with changes in muscle fiber diameter and miRNAs expression. There was an increase in the mRNA levels of catabolic pathways components (FBXO25, ATG12, BCL2) and energetic metabolism-related genes (PGC1αand SDHA), together with a decrease in PPARβ/δmRNA levels. Interestingly, an increase in mRNA levels of anabolic genes (PI3K and mTORC1 complex: mTOR, mLST8 and RAPTOR) was also observed during food restriction. After refeeding, muscle morphology showed similar patterns of the control group; the majority of genes were slightly up- or down-regulated in fast and slow muscle, respectively; the levels of all miRNAs increased in fast muscle and some of them decreased in slow muscle. Our findings demonstrated that a short period of food restriction in juvenile pacu had a considerable impact on fast muscle, increasing the expression of anabolic (PI3K and mTORC1 complex: mTOR, mLST8 and RAPTOR) and energetic metabolism genes. The miRNAs (miR-1, miR-206, miR-199 and miR-23a) were more expressed during refeeding and while their target genes (IGF-1, mTOR, PGC1α and MAFbx), presented a decreased expression. The alterations in mTORC1 complex observed during fasting may have influenced the rates of protein synthesis by using amino acids from protein degradation as an alternative mechanism to preserve muscle phenotype and metabolic demand maintenance.
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Palavras-chave
Músculo esquelético, Jejum, Realimentação, Pacu