M. Cagnazzo

Comparison of prenatal muscle tissue expression profiles of two pig breeds differing in muscle characteristics.

The objective of this study was to compare purebred Duroc and Pietrain prenatal muscle tissue transcriptome expression levels at different stages of prenatal development to gain insight into the differences in muscle tissue development in these pig breeds. Commercial western pig breeds have been selected for muscle growth for the past 2 decades. Pig breeds differ for their muscle phenotypes (i.e., myofiber numbers and myofiber types). Duroc and Pietrain pig breeds are extremes; Duroc pigs have redder muscle fiber types with more intramuscular fat, and Pietrain pigs have faster-growing and whiter muscle fiber types. Pietrain pigs are more muscular than Duroc pigs, whereas Duroc pigs are fatter than Pietrain pigs. The genomic background underlying these breed-specific differences is poorly known. Myogenesis is a complex exclusive prenatal process involving proliferation and differentiation (i.e., fusion) of precursor cells called myoblasts. We investigated the difference in the prenatal muscle-specific transcriptome profiles of Duroc and Pietrain pigs using microarray technology. The microarray contained more than 500 genes affecting myogenesis, energy metabolism, muscle structural genes, and other genes from a porcine muscle cDNA library. The results indicated that the expression of the myogenesis-related genes was greater in early Duroc embryos than in early Pietrain embryos (14 to 49 d of gestation), whereas the opposite was found in late embryos (63 to 91 d of gestation). These findings suggest that the myogenesis process is more intense in early Duroc embryos than in Pietrain embryos but that myogenesis is more intense in late Pietrain fetuses than in Duroc fetuses. Transcriptomes of muscle structural genes followed that pattern. The energy metabolism genes were expressed at a higher level in prenatal Pietrain pigs than in prenatal Duroc pigs, except for d 35, when the opposite situation was found. Fatty acid metabolism genes were expressed at a higher level in early (14 to 49 d of gestation) Duroc embryos than in Pietrain embryos. Better understanding of the genomic regulation of tissue formation leads to improved knowledge of the genome under selection and may lead to directed breed-specific changes in the future.

Quantitative Real Time PCR approach to study gene expression profile during prenatal growth of skeletal muscle in pig of Duroc and Pietrain breeds

Riassunto Applicazione della real time PCR allo studio dell’espressione genica nel tessuto muscolare scheletrico di suino durante lo sviluppo embrionale nelle razze Duroc e Pietrain. La PCR quantitativa “real time”, è stata utilizzata per analizzare i livelli d’espressione di geni muscolari di suino durante lo sviluppo embrionale. A tale scopo è stato utilizzato il LightCycler™system per quantificare il numero di copie di mRNA di geni del tessuto muscolare scheletrico e confrontarne il pattern di espressione nelle razze suine Duroc e Pietrain, molto diverse per caratteristiche produttive e per la qualità della carne e della carcassa. Il livello d’espressione di questi geni è stato monitorato con quantificazione relativa, basata sul rapporto tra il numero copie cDNA/µl di ciascun gene target rispetto al gene di riferimento β2-microglobulina (B2M).