title: Short-hairpin Mediated Myostatin Knockdown Resulted in Altered Expression of Myogenic Regulatory Factors with Enhanced Myoblast Proliferation in Fetal Myoblast Cells of Goats.
abstract: Myostatin (MSTN) is a well-known negative regulator of skeletal muscle development. Reduced expression due to natural mutations in the coding region and knockout as well as knockdown of MSTN results in an increase in the muscle mass. In the present study, we demonstrated as high as 60 and 52% downregulation (p < 0.01) of MSTN mRNA and protein in the primary fetal myoblast cells of goats using synthetic shRNAs (n = 3), without any interferon response. We, for the first time, evaluated the effect of MSTN knockdown on the expression of MRFs (namely, MyoD, Myf5), follistatin (FST), and IGFs (IGF-1 & IGF-2) in goat myoblast cells. MSTN knockdown caused an upregulation (p < 0.05) of MyoD and downregulation (p < 0.01) of MYf5 and FST expression. Moreover, we report up to ∼four fold (p < 0.001) enhanced proliferation in myoblasts after four days of culture. The anti-MSTN shRNA demonstrated in the present study could be used for the production of transgenic goats to increase the muscle mass.
authors.affiliation: a Genome Analysis Laboratory, Animal Genetics Division , ICAR- Indian Veterinary Research Institute , Izatnagar , Bareilly , India.
authors.forename: Rohit
authors.initials: R
authors.lastname: Kumar
authors.affiliation: a Genome Analysis Laboratory, Animal Genetics Division , ICAR- Indian Veterinary Research Institute , Izatnagar , Bareilly , India.
authors.forename: Satyendra Pal
authors.initials: SP
authors.lastname: Singh
authors.affiliation: a Genome Analysis Laboratory, Animal Genetics Division , ICAR- Indian Veterinary Research Institute , Izatnagar , Bareilly , India.
authors.forename: Abhijit
authors.initials: A
authors.lastname: Mitra
mesh_terms: D000818:Animals; D049109:Cell Proliferation / Q000502:physiology*; D002478:Cells, Cultured; D015536:Down-Regulation / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D055785:Gene Knockdown Techniques / Q000662:veterinary; D006041:Goats; D024510:Muscle Development / Q000235:genetics*; D032446:Myoblasts / Q000166:cytology / Q000502:physiology*; D018006:Myogenic Regulatory Factors / Q000235:genetics / Q000378:metabolism*; D055435:Myostatin / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*
chemical_list: D018006:Myogenic Regulatory Factors; D055435:Myostatin; D034741:RNA, Small Interfering
keywords: Anti-myostatin shRNA; IGFs; MRFs; caprine fetal myoblasts; interferon response