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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.2016.tde-05012016-154015
Document
Author
Full name
Berna Inés Giménez Kappeler
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 2015
Supervisor
Committee
Coutinho, Luiz Lehmann (President)
Ament, Andrezza Maria Felicio
Regitano, Luciana Correia de Almeida
Title in Portuguese
Identificação de microRNAs envolvidos com a maciez da carne em bovinos da raça Nelore
Keywords in Portuguese
Bos indicus
Longissumus dorsi
Expressão diferencial
Força de cisalhamento
miRDeep2
QuasiSeq
Redes
Abstract in Portuguese
O Brasil ocupa a segunda posição mundial na produção de carne bovina, a implantação de novas ferramentas para a seleção de animais zebuínos (Bos indicus) com carne de melhor qualidade tem uma importante contribuição para a competividade da pecuária de corte. Neste contexto, compreender os padrões de expressão de microRNAs específicos envolvidos nos processos que afetam a maciez da carne é fundamental para a sua produção, uma vez que essa característica organoléptica é de grande valor na aceitação deste alimento pelos consumidores. O advento das tecnologias de sequenciamento de nova geração em conjunto com o uso de ferramentas de bioinformática tem permitido o estudo do genoma em larga escala de forma mais rápida e com menor custo. Para este estudo, foram utilizadas amostras do músculo Longissimus dorsi de 34 animais da raça Nelore com medidas extremas de valor genético estimado (EBV) para força de cisalhamento (FC). Os RNAs totais foram extraídos, as bibliotecas de microRNA foram construídas e os sequenciamentos foram realizados em equipamento da plataforma Illumina (MiSeq). O processamento dos dados foi feito por meio dos softwares FastQC, Cutadapt e miRDeep2 e as análises de expressão diferencial foram realizadas por meio do programa estatístico QuasiSeq. Utilizando um critério de taxa de descoberta de falsos positivos (FDR) inferior a 0,1, três microRNAs (bta-mir-182, bta-mir-183, bta-mir-338) foram identificados como diferencialmente expressos entre os grupos de animais com valores extremos de EBV para FC. Um total de 1204 genes alvos foi previsto e análises funcionais de enriquecimento foram realizadas por ferramentas de bioinformática. Várias redes e vias metabólicas como a sinalização de apoptose e regulação dos mecanismos celulares pela protease calpaína foram obtidas, demonstrando assim que os genes alvos identificados estariam envolvidos em muitos processos metabólicos relacionados com a maciez da carne bovina.
Title in English
Identification of microRNAs involved in meat tenderness in Nellore cattle
Keywords in English
Bos indicus
Differential expression
Longissumus dorsi
miRDeep2
Network
QuasiSeq
Shear force
Abstract in English
Brazil occupies the second world position in beef production and thus, the implementation of new tools to select zebuine animals (Bos indicus) with better beef quality has an important contribution to the competitiveness of beef cattle. Inside this context, to comprehend the microRNAs expression patterns involved in the processes that are related to beef tenderness is essential to the meat production since this organoleptic characteristic has a high value in meat acceptance by the consumers. The advent of new generation sequencing technologies along with the biotechnology tools usage has allowed large-scale genome studies as well as faster and cheaper analysis. In this study, samples of the Longissimus dorsi muscle from 34 animals of Nellore cattle breed with extreme estimated genetic value (EBV) for shear force (FC) were used. The total RNAs were extracted, libraries of microRNA were built and finally the sequencing was performed using the Illumina (MiSeq) platform equipment. Data processing was done using FastQC, Cutadapt and miRDeep2 softwares while the differential expression analyzes were realized through the statistical package QuasiSeq. Using a false discovery rate (FDR) criteria below 0.1, three microRNAs (bta-mir- 182, bta-mir-183, bta-mir-338) were identified as differentially expressed among the group of animals with extreme EBV values for FC. A total of 1024 target genes were predicted and functional analyzes of enrichment were performed using bioinformatics tools. Many metabolic networks and pathways such as the apoptosis signalization and cell regulation mechanisms by calpain protease were obtained, demonstrating therefore that the identified target genes would be involved in many metabolic processes related with the beef tenderness.
 
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Publishing Date
2016-01-18
 
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