title: The effect of myostatin silencing by lentiviral-mediated RNA interference on goat fetal fibroblasts.
abstract: Myostatin is a transforming growth factor-β family member that acts as a negative regulator of skeletal muscle mass. To identify possible myostatin inhibitors that may promote muscle growth, we used RNA interference mediated by a lentiviral vector to knockdown myostatin in goat fetal fibroblast cells. We also investigated the expression changes in relevant myogenic regulatory factors (MRFs) and adipogenic regulatory factors in the absence of myostatin in goat fetal fibroblasts. Quantitative RT-PCR revealed that myostatin transcripts were significantly reduced by 75 % (P < 0.01). Western blot showed that myostatin protein expression was reduced by 95 % (P < 0.01). We also found that the mRNA expression of activin receptor IIB (ACVR2B) significantly increased by 350 % (P < 0.01), and p21 increased 172 % (P < 0.01). Furthermore, myostatin inhibition decreased Myf5 and increased MEF2C mRNA expression in goat fetal fibroblasts, suggesting that myostatin regulates MRFs differently in fibroblasts compared to muscle. In addition, the expression of adipocyte marker genes peroxisome proliferator-activated receptor (PPAR) γ and leptin, but not CCAAT/enhance-binding protein (C/EBP) α and C/EBPβ, were upregulated at the transcript level after myostatin silencing. These results suggest that we have generated a novel way to block myostatin in vitro, which could be used to improve livestock meat production and gene therapy of musculoskeletal diseases. This also suggests that myostatin plays a negative role in regulating the expression of adipogenesis related genes in goat fetal fibroblasts.
authors.affiliation: National Center for Molecular Genetics and Breeding of Animal, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, People's Republic of China.
authors.forename: Jian
authors.initials: J
authors.lastname: Lu
authors.forename: Caihong
authors.initials: C
authors.lastname: Wei
authors.forename: Xiaoning
authors.initials: X
authors.lastname: Zhang
authors.forename: Lingyang
authors.initials: L
authors.lastname: Xu
authors.forename: Shifang
authors.initials: S
authors.lastname: Zhang
authors.forename: Jiasen
authors.initials: J
authors.lastname: Liu
authors.forename: Jiaxue
authors.initials: J
authors.lastname: Cao
authors.forename: Fuping
authors.initials: F
authors.lastname: Zhao
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.forename: Bichun
authors.initials: B
authors.lastname: Li
authors.forename: Lixin
authors.initials: L
authors.lastname: Du
mesh_terms: D030301:Activin Receptors, Type II / Q000235:genetics / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics / Q000378:metabolism; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics / Q000378:metabolism; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000235:genetics / Q000378:metabolism; D005333:Fetus / Q000166:cytology*; D005347:Fibroblasts / Q000378:metabolism*; D005786:Gene Expression Regulation; D006041:Goats / Q000196:embryology*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D020738:Leptin / Q000235:genetics / Q000378:metabolism; D009132:Muscles / Q000378:metabolism; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D009928:Organ Specificity / Q000235:genetics; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D051900:Smad3 Protein / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; D022782:CCAAT-Enhancer-Binding Protein-beta; D050759:Cyclin-Dependent Kinase Inhibitor p21; D020738:Leptin; D055435:Myostatin; D047495:PPAR gamma; D034741:RNA, Small Interfering; D051900:Smad3 Protein; D030301:Activin Receptors, Type II; C425584:activin receptor type II-B