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The aim of the present study was to define the effect of TGF-β1 on C2C12 myoblasts myogenesis. TGF-β1 together with its receptor is a negative auto-paracrine regulator of myogenesis, which influences the proliferation, differentiation, and functions of muscle cells. TGF-β1 exerts highly significant inhibitory effect on differentiation of C2C12 mouse myoblasts manifested by the impairment of cell fusion and very low expression of myosin heavy chain. The study of differentiating C2C12 mouse myoblasts treated with TGF-β1 revealed 502 genes (436 down-regulated and 66 up-regulated) with statistically different expression. TGF-β1-regulated genes were identified to be involved in 29 biological processes, 29 molecular functions groups and 59 pathways. The strongest inhibiting effect of TGF-β1 was observed in the cadherin and Wnt pathways. The key-genes that could play the role of TGF-β1 targets during myoblasts differentiation was identified such as: Max, Creb1, Ccna2, Bax, MdfI, Tef, Tubg1, Cxcl5, Rho, Calca and Lgals4.
Experiments were performed on 180 mice from two lines dubbed light (L) and heavy (C), selected divergently for body weight over 108 generations. The main hypothesis was that the changes occurring in body weight and muscle weight as a result of directed divergent selection could be associated with changes in the transcription of some miogenic genes and/or with the level of proteins regulating myogenesis and with the composition of muscle fibres.Hind limb muscle masses from females and males of the two lines were weighed at 1 and 3 weeks and 3 month of age. Morphological analysis for histological cross-sections of the gastroenemius muscle was carried out in 3-week and 3-month-old mice. The percentage comparison of nuclei in muscle fibres were analysed, too. Levels of MYOD1, MYF-5 and myogenin at the same time points were determined using Western blotting. Microsatellite markers for MyoD1, Myf-5 and MYOG were used to compare allele frequencies of analysed genes in both lines. There were differences in muszle weight between the sexes at age of 3 months. Muscles of the hind limbs were heavier in males than in females by 23.7% in line C, and by 14% in line L. Significant differences in muscle mass were accompanied by changes in muscle fibre size. The number of large-diameter muscle fibres increased with animals’ age, and in females fibres of diameters of 60-80 μm accounted for 38% of the total in line C, as compared with 94% preponderance of smaller (20-60 μm) fibres in the muscle of line L females. The numbers of nuclei were clearly greater in line C than in line L individuals, as well as in 3-month-old animals as compared with those at 3 weeks of age.Selection have brought about change, not only in myogenesis, but also in the frequency of alleles of microsatellite markers MyoD1, Myf-5 and at the myogenin locus, thus suggesting that molecular differences between the lines have arisen. Differences in the levels of MYOD1, MYF5 and MYOG are evident between both sexes and the selected lines of mice.
Pre- and postnatal processes determine the final outcome of breeding of pigs in terms of traits related to carcass and meat quality at slaughter. In particular, the number of myofibers and to a large extent their metabolic and contractile properties, which also influence their size, are determined prenatally during the process of myogenesis. By this, postnatal muscle growth and parameters of meat quality are modulated. The metabolic balance, biochemical and biophysical preslaughter properties of muscle prior to slaughter determine the process of maturation of muscle to meat.Thus, differential regulation of the abundance of transcripts of biological networks in prenatal and postnatal muscle affect biochemical processes of meat maturation. In general, because the traits of interest are typically not expressed at prenatal stages, no direct relationship between phenotype and gene expression pattern can be established. However, trait-related differential expression within any prenatal developmental stage can be assessed based on known estimated breeding values, known QTL-genotypes and/or based on breed differences. Expression profiles of muscle at slaughter can directly be linked to meat quality. A suitable experimental design of “matched samples” is the discordant sib pair design. Here it is exemplarily discussed that differentially expressed transcript profiles of M. longissimus dorsi at prenatal and postnatal stages offer an insight into the biological processes in the live muscle that affect the process of meat maturation and finally meat quality.
MYOG and MYF6 belong to the MyoD gene family. They code for the bHLH transcription factors playing a key role in later stages of myogenesis: differentiation and maturation of myotubes. Three SNPs in porcine MYF6 and two in porcine MYOG were analysed in order to establish associations with chosen carcass quality and growth rate traits in Polish Landrace, Polish Large White and line 990 sows. No statistically significant effect of SNP in the promoter region of the MYF6 gene on its expression measured on mRNA level was found. Associations between the genotype at the MYF6 locus and carcass quality traits appeared to be breed-dependent. The C allele in the case of SNP in the promoter region and GC haplotype in exon 1 were advantageous for right carcass side weight in Polish Landrace sows and disadvantageous for this trait in Polish Large White sows. These gene variants were also the most advantageous for loin and ham weight in sows of line 990. The mutation in exon 1 of the MYOG gene had no statistically significant association with carcass quality traits and the mutation in the 3’-flanking region had the breed-dependent effect as well. These results suggest that SNPs analysed in this study are not causative mutations, but can be considered as markers of some other, still unrevealed genetic polymorphism that influences the physiological processes and phenotypic traits considered in this study.
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