Meta-análise integrativa da expressão gênica global no músculo esquelético de peixes
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Data
2022-02-23
Autores
Perez, Érika Stefani
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O crescimento muscular dos peixes, o qual desperta grande interesse do setor produtivo, é regulado por múltiplos genes que fazem deste, um complexo processo ainda pouco compreendido. Nesse contexto, a reanálise do perfil de mRNAs é uma estratégia significativa que permite a compreensão do crescimento desse tecido. Com base nisso, os objetivos deste estudo foram reanalisar, integrar e comparar o perfil de mRNAs do músculo esquelético de peixes, bem como encontrar possíveis genes alvos envolvidos com o crescimento e manutenção do fenótipo muscular. Foi realizada uma meta-análise utilizando palavras-chave no banco de dados GEO Datasets e European Nucleotide Archive. Os dados de microarray foram reanalisados e partir dos genes diferencialmente expressos (GDEs) de cada dataset foi realizada uma análise individual que identificou as vias mais comuns dos estudos (p<0.05). Essas vias, presentes em pelo menos dois datasets, tiveram seus genes selecionados para a construção da rede de interação molecular. Com base nessa rede, foi obtida a importante relação entre ribossomos e ciclo celular em que alguns genes foram selecionados para investigação através de RT-qPCR: rps27a, eef1a2, ccnd1, cdkn1a e pcna, além da mtor. As investigações foram realizadas em pacus (P.mesopotamicus) de 12g submetidos ao jejum de 4 dias (J4d) e realimentação de 3 dias (R3d), e em pacus com 30g submetidos ao jejum de 15 dias (J15d) e realimentação de 6 horas (R6h) e 15 dias (R15d). O período mais curto de jejum (J4d) promoveu um aumento na expressão da mtor. A expressão de rps27a, ccnd1, cdkn1a e do pcna foi reduzida em R3d comparada ao J4d. No período mais prolongado de jejum e realimentação, a expressão da mtor foi reduzida em R15d em comparação à R6h, assim como do pcna. Considerando a expressão de rps27a, houve uma diminuição em R15d comparado ao grupo J15d, enquanto a expressão de eef1a2 não se alterou. A expressão de ccnd1 foi elevada em J15d e R6h em comparação aos grupos C e R15d. Alterações importantes também foram observadas na expressão da cdkn1a que manteve altos níveis no grupo Controle (C) e J15d e diminuição em R6h e R15d. Considerando a importante relação entre os ribossomos e o ciclo celular obtida pela meta-análise, a investigaçãodesses genes selecionados demonstra estratégias adotadas pela musculatura que inclui a manutenção da capacidade traducional, sustentada pela biogênese dos ribossomos, a otimização da polimerização proteica e o reparo do tecido diante do estresse decorrente da falta de alimento. Além disso, considerando o ciclo-celular, a expressão de seus reguladores é modulada frente à redução e ao reestabelecimento das condições pró-proliferação, visando a sobrevivência celular durante o estresse nutricional. Dessa maneira, nosso estudo pode fornecer novos insights acerca da importância dos genes que regulam a tradução, transcrição e replicação, como marcadores associados à manutenção do fenótipo e função muscular, visando o crescimento dos peixes.
The muscle growth of fish, which arouses great interest in the productive sector, is regulated by multiple genes that make this a complex process that is still poorly understood. In this context, the reanalysis of the mRNAs profile is a significant strategy that allows the understanding of the growth of this tissue. Based on this, the objectives of this study were to reanalyze, integrate and compare the fish skeletal muscle mRNA profile, as well as find possible target genes involved in the growth and maintenance of the muscle phenotype. A meta-analysis was performed using keywords in the GEO Datasets and European Nucleotide Archive database. The microarray data were reanalyzed and from the differentially expressed genes (GDEs) of each dataset, an individual analysis was performed that identified the most common pathways in the studies (p<0.05). These pathways, present in at least two datasets, had their genes selected for the construction of the molecular interaction network. Based on this network, the important relationship between ribosomes and cell cycle was obtained, in which some genes were selected for investigation through RT-qPCR: rps27a, eef1a2, ccnd1, cdkn1a and pcna, in addition to mtor. Investigations were performed in 12g pacus (P.mesopotamicus) submitted to 4 days fasting (J4d) and 3 days refeeding (R3d), and in 30g pacu submitted to 15 days fasting (J15d) and 6 hours refeeding (R6h) and 15 days (R15d). The shorter period of fasting (J4d) promoted an increase in mtor expression. The expression of rps27a, ccnd1, cdkn1a and pcna was reduced in R3d compared to J4d. In the longer period of fasting and refeeding, mtor expression was reduced in R15d compared to R6h, as well as pcna. Considering the expression of rps27a, there was a decrease in R15d compared to the J15d group, while the expression of eef1a2 did not change. The expression of ccnd1 was elevated in J15d and R6h compared to groups C and R15d. Important changes were also observed in cdkn1a expression, which maintained high levels in the Control (C) and J15d groups and decreased in R6h and R15d. Considering the important relationship between the ribosomes and the cell cycle obtained by the meta-analysis, the investigation of these selected genes demonstrates strategies adopted by musculature that include the maintenance of the translational capacity, supported by the biogenesis of the ribosomes, the optimization of protein polymerization and the repair of the tissue in the face of stress resulting from lack of food. Furthermore, considering the cell cycle, the expression of its regulators is modulated against the reduction and reestablishment of pro-proliferation conditions, aiming at cell survival during nutritional stress. In this way, our study may provide new insights into the importance of genes that regulate translation, transcription and replication, as markers associated with the maintenance of phenotype and muscle function, targeting fish growth.
The muscle growth of fish, which arouses great interest in the productive sector, is regulated by multiple genes that make this a complex process that is still poorly understood. In this context, the reanalysis of the mRNAs profile is a significant strategy that allows the understanding of the growth of this tissue. Based on this, the objectives of this study were to reanalyze, integrate and compare the fish skeletal muscle mRNA profile, as well as find possible target genes involved in the growth and maintenance of the muscle phenotype. A meta-analysis was performed using keywords in the GEO Datasets and European Nucleotide Archive database. The microarray data were reanalyzed and from the differentially expressed genes (GDEs) of each dataset, an individual analysis was performed that identified the most common pathways in the studies (p<0.05). These pathways, present in at least two datasets, had their genes selected for the construction of the molecular interaction network. Based on this network, the important relationship between ribosomes and cell cycle was obtained, in which some genes were selected for investigation through RT-qPCR: rps27a, eef1a2, ccnd1, cdkn1a and pcna, in addition to mtor. Investigations were performed in 12g pacus (P.mesopotamicus) submitted to 4 days fasting (J4d) and 3 days refeeding (R3d), and in 30g pacu submitted to 15 days fasting (J15d) and 6 hours refeeding (R6h) and 15 days (R15d). The shorter period of fasting (J4d) promoted an increase in mtor expression. The expression of rps27a, ccnd1, cdkn1a and pcna was reduced in R3d compared to J4d. In the longer period of fasting and refeeding, mtor expression was reduced in R15d compared to R6h, as well as pcna. Considering the expression of rps27a, there was a decrease in R15d compared to the J15d group, while the expression of eef1a2 did not change. The expression of ccnd1 was elevated in J15d and R6h compared to groups C and R15d. Important changes were also observed in cdkn1a expression, which maintained high levels in the Control (C) and J15d groups and decreased in R6h and R15d. Considering the important relationship between the ribosomes and the cell cycle obtained by the meta-analysis, the investigation of these selected genes demonstrates strategies adopted by musculature that include the maintenance of the translational capacity, supported by the biogenesis of the ribosomes, the optimization of protein polymerization and the repair of the tissue in the face of stress resulting from lack of food. Furthermore, considering the cell cycle, the expression of its regulators is modulated against the reduction and reestablishment of pro-proliferation conditions, aiming at cell survival during nutritional stress. In this way, our study may provide new insights into the importance of genes that regulate translation, transcription and replication, as markers associated with the maintenance of phenotype and muscle function, targeting fish growth.
Descrição
Palavras-chave
Meta-análise, Peixes, Crescimento do músculo esquelético, Meta-analysis, Fish, Skeletal muscle growth