Efeito da desmama precoce suplementada com concentrado sobre a expressão gênica do longissimus thoracis em bezerros da raça nelore
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Data
2023-04-17
Autores
Pedro, Ariane Enara
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Universidade Estadual Paulista (Unesp)
Resumo
Este estudo foi conduzido com bezerros da raça Nelore pertencentes a dois grupos diferentes, desmama precoce (120 dias) e desmama convencional (205 dias), objetivando analisar de forma global as diferenças na expressão gênica muscular (Longissimus thoracis, LT) entre amostras de tecido muscular coletadas por meio do sequenciamento do RNA mensageiro utilizando a técnica de RNA-Seq e avaliar a relação entre genes diferencialmente expressos sobre suas funções gênicas enriquecidas e vias metabólicas das fibras de tecido muscular esquelético em bezerros Nelore. Para isso, foram utilizados dezesseis bezerros, dos quais oito foram desmamados precocemente e oito do desmama convencional. Os bezerros desmamados precocemente foram suplementados até duzentos e cinco dias com uma dieta contendo 20% de Proteína Bruta e 75% de NDT, em uma quantidade de 20g de MS/kg de PV, enquanto os desmamados aos duzentos e cinco dias permaneceram a pasto sob aleitamento. Para comparação da expressão gênica entre tratamentos foi realizado o RNA Seq de alíquotas de musculo Longissimus thoracis coletadas por biópsia aos duzentos e cinco dias dos dezesseis indivíduos. As análises de enriquecimento funcional foram realizadas por meio de teste hipergeométrico para super-representação de genes diferencialmente expressos em vias metabólicas KEGG utilizando o pacote clusterProfiler do programa R, pela metodologia do pacote edgeR. Um total de 14.076 genes estavam presentes, visto que 762 DEGs enriquecidos significativos foram observados após o ajuste de FDR com valor de p ajustado <0,05. Entre os 762 termos de GO, 405 são genes up-regulated e 357 genes down-regulated pertenciam ao processo biológico envolvidos no metabolismo lipídico nos respectivos momentos. Foram selecionados com base nos resultados das análises os primeiros 20 genes regulados positivamente com atuação no metabolismo lipídico em animais desmamados precocemente sendo eles: ADIPOQ, FABP4, FADS1, FADS2, SCD, ACSM1, ELOVL6, HSD17B12, PCK2, PER2, SREBF1, FBP1, NR4A3, PRKAG3, ACACA, FASN, COL4A3, ADCY1 e C8G. Os resultados de vias KEGG comuns com método e Vias KEGG enriquecidas a partir de DEGs comuns indicaram forte regulação positiva de genes relacionados a adipogênese e lipogênese e regulação negativa da degradação e ácidos graxos no tecido muscular em bezerros que foram desmamados aos cento e vinte dias.
This study was conducted with Nelore calves belonging to two different groups, early weaning (120 days) and conventional weaning (205 days), aiming to analyze globally the differences in muscle gene expression (Longissimus thoracis, LT) between muscle tissue samples collected by messenger RNA sequencing using the RNA-Seq and evaluate the relationship between differentially expressed genes on their enriched gene functions and metabolic pathways of skeletal muscle tissue fibers in Nelore calves. For this, sixteen calves were used, of which eight were weaned early and eight from conventional weaning. Calves weaned early were supplemented until two hundred and five days with a diet containing 20% crude protein and 75% NDT, in an amount of 20g DM/kg BW, while calves weaned at two hundred and five days remained at pasture under suckling. For comparison of gene expression between treatments, RNA Seq of aliquots of Longissimus thoracis muscle collected by biopsy at two hundred and five days from the sixteen individuals was performed. Functional enrichment analyses were performed by hypergeometric test for over-representation of differentially expressed genes in KEGG metabolic pathways using the clusterProfiler package of the R program, by the edgeR package methodology. A total of 14,076 genes were present, as 762 significant enriched DEGs were observed after FDR adjustment with adjusted p-value <0.05. Among the 762 GO terms, 405 are upregulated genes and 357 down-regulated genes belonged to the biological process involved in lipid metabolism at the respective times. Based on the analysis results, 20 positively regulated genes acting on lipid metabolism in early weaned animals were selected, as follows: ADIPOQ, FABP4, FADS1, FADS2, SCD, ACSM1, ELOVL6, HSD17B12, PCK2, PER2, SREBF1, FBP1, NR4A3, PRKAG3, ACACA, FASN, COL4A3, ADCY1 and C8G. The results of common KEGG pathways with method and enriched KEGG pathways from common DEGs indicated strong positive regulation of genes related to adipogenesis and lipogenesis and negative regulation of degradation and fatty acids in muscle tissue in calves that were weaned at one hundred and twenty days.
This study was conducted with Nelore calves belonging to two different groups, early weaning (120 days) and conventional weaning (205 days), aiming to analyze globally the differences in muscle gene expression (Longissimus thoracis, LT) between muscle tissue samples collected by messenger RNA sequencing using the RNA-Seq and evaluate the relationship between differentially expressed genes on their enriched gene functions and metabolic pathways of skeletal muscle tissue fibers in Nelore calves. For this, sixteen calves were used, of which eight were weaned early and eight from conventional weaning. Calves weaned early were supplemented until two hundred and five days with a diet containing 20% crude protein and 75% NDT, in an amount of 20g DM/kg BW, while calves weaned at two hundred and five days remained at pasture under suckling. For comparison of gene expression between treatments, RNA Seq of aliquots of Longissimus thoracis muscle collected by biopsy at two hundred and five days from the sixteen individuals was performed. Functional enrichment analyses were performed by hypergeometric test for over-representation of differentially expressed genes in KEGG metabolic pathways using the clusterProfiler package of the R program, by the edgeR package methodology. A total of 14,076 genes were present, as 762 significant enriched DEGs were observed after FDR adjustment with adjusted p-value <0.05. Among the 762 GO terms, 405 are upregulated genes and 357 down-regulated genes belonged to the biological process involved in lipid metabolism at the respective times. Based on the analysis results, 20 positively regulated genes acting on lipid metabolism in early weaned animals were selected, as follows: ADIPOQ, FABP4, FADS1, FADS2, SCD, ACSM1, ELOVL6, HSD17B12, PCK2, PER2, SREBF1, FBP1, NR4A3, PRKAG3, ACACA, FASN, COL4A3, ADCY1 and C8G. The results of common KEGG pathways with method and enriched KEGG pathways from common DEGs indicated strong positive regulation of genes related to adipogenesis and lipogenesis and negative regulation of degradation and fatty acids in muscle tissue in calves that were weaned at one hundred and twenty days.
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Expressão gênica, RNA sequence, Bovino metabolismo