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Myostatin (MSTN) and transforming growth factor-ß1 (TGF-ß1) belong to the same TGF-ß superfamily of proteins. They are involved in regulation of skeletal muscle growth and development as well as muscle catabolism. The aim of the present study was to investigate the relationship between mstn and TGF-ß1 expression in proliferating and differentiating mouse C2C12 myoblasts cultured in normal and catabolic conditions and to evaluate the effect of exogenous TGF-ß1 as well as "knock down" of TGF-ß1 receptor type II on mstn expression in proliferating and differentiating myogenic cells. The direct effect of TGF-ß1 on myostatin was also examined. Myostatin expression increased gradually with cell confluency in proliferating cultures, while the level of TGF-ß1, detected in the form of a 100 kDa small latent complex diminished. Myostatin expression was accompanied by a partial cell cycle arrest. Three forms of myostatin were found: a 52 kDa precursor, a 40 kDa latency associated propeptide, and a 26 kDa active peptide. A decrease in myostatin and TGF-ß1 levels was observed during the first three days of differentiation, which was subsequently followed by significant increase of their expression during next three to four days of differentiation. Catabolic state induced by dexamethasone significantly increased the level of all forms of myostatin as well as latent (100 kDa) and active (25 kDa) forms of TGF-ß1 in differentiating myoblasts in a dose dependent manner. Exogenous TGF-ß1 (2 ng/ml) significantly increased myostatin levels both in proliferating and differentiating C2C12 myoblasts, whereas silencing of the TGF-ß1 receptor II gene significantly lowered myostatin level in examined cells. The presented results indicate that TGF-ß1 may control myostatin-related regulation of myogenesis through up-regulation of myostatin, predominantly in the course of terminal differentiation and glucocorticoid-dependent catabolic stimulation.
Myostatin (GDF-8) is a key protein responsible for skeletal muscle growth and development so mutations in the mstn gene can have major economic and breeding consequences. The aim of the present study was to investigate polymorphism in the 5’flanking region of the mstn gene and its possible influence on the myostatin level in skeletal muscles of Polish Black-and-White bulls. The relation between expression of myostatin and another member of the TGF-β superfamily, TGF-β1, was also examined. We uncovered polymorphism in the 5’flanking region of the mstn gene: G/C substitution at position -7828 (relative to translation start codon ATG). The most frequent genotype was GC (43.6%), followed by genotypes CC (34.2%) and GG (22.2%). The concentration of the active form of myostatin (26 kDa) in M. semitendinosus of homozygotes (CC) was the lowest, whereas the expression of this cytokine in heterozygotes was the highest. The changes in mstn expression did not, however, influence average carcass traits (weight of valuable cuts and weight of lean in valuable cuts). The pattern of differences in the TGF-β1 concentration corresponded to that observed for myostatin.
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