MiR-29b-1-5p regulates the proliferation and differentiation of chicken primary myoblasts and analysis of its effective targets

Abstract

Several recent studies investigated the role of the miR-29 family in muscle development. However, only a few studies focused on chicken skeletal muscle. In the present study, cell cycle, 5-ethynyl-2’-deoxyuridine ( EdU ), cell counting kit-8 ( CCK-8 ), and other assays indicated that miR-29b-1-5p can inhibit the proliferation of chicken primary myoblasts ( CPMs ); the western blot assay and immunofluorescence detection of MYHC demonstrated that miR-29b-1-5p can promote the differentiation of myoblasts. The functional enrichment analysis revealed that the target genes of miR-29b-1-5p may be involved in muscle tissue development, muscle organ development, and striated muscle tissue development, which are biological processes related to muscle development. The correlation analysis showed that these 6 genes, that is, ankyrin repeat domain 9 ( ANKRD9 ), lactate dehydrogenase A ( LDHA ), transcription factor 12 ( TCF12 ), FAT atypical cadherin 1 ( FAT1 ), lin-9 homolog ( LIN9 ), and integrin beta 3 binding protein ( ITGB3BP ), can be used as effective candidate target genes of miR-29b-1-5p. Moreover, miR-29b-1-5p inhibits the expression of ANKRD9 by directly binding the 3′UTR of ANKRD9 . Overall, these data indicate that miR-29b-1-5p inhibits the proliferation of primary chicken myoblasts, stimulates their differentiation, and is involved in the process of muscle development and that its effective target gene is ANKRD9 . This study identified the molecular mechanism of miR-29b-1-5p in chicken muscle development.

MiRNA sequencing of Embryonic Myogenesis in Chengkou Mountain Chicken

“…A E12: gga-miR-130b-5p target genes; B E16: gga-miR-1643-5p target genes; C E19: gga-miR-12218-5p target genes; D E21: gga-miR-132b-5p target genes gga-miRNA-454-3p does not affect primary myoblast differentiation, it inhibits differentiation by targeting the myotube-associated protein SBF2 [27]. A study focused on the chicken skeletal muscle indicated that miR-29b-1-5p inhibits the proliferation of chicken primary myoblasts and promotes the differentiation of myoblasts via an effective target gene, ANKRD9 [28]. Meanwhile, miR-29b-1-5p and miR-133a-5p are sponges for circFGFR2 in skeletal muscle proliferation and differentiation [29].…”

Mining of chicken muscle growth genes and the function of important candidate gene RPL3L in muscle development

“…For example, muscle phosphofructo-1 kinase ( PFKM ) is a key regulatory enzyme of glycolysis ( Garcia et al, 2009 ), and regulates the insulin-stimulated glycolytic pathway in chicken skeletal muscle ( Seki et al, 2006 ). Lactate dehydrogenase A ( LDHA ) inhibits the proliferation and promotes differentiation of chicken primary myoblasts through being targeted by miR-29b-1-5p ( Li et al, 2022 ). Studies have pointed out that white streaks and lignified chicken breasts in broiler breasts may be related to LDHA , but no causal relationship has been established ( Malila et al, 2019 ).…”

RRM2 promotes the proliferation of chicken myoblasts, inhibits their differentiation and muscle regeneration

“…CPMs were isolated from the leg muscles of E11 AA broiler following previously described methods ( Cai et al, 2018 ; Li et al, 2022 ; Wei et al, 2022 ). Each in vitro cell experiment was isolated and cultured from at least 10 E11 AA broiler leg muscles.…”

“…After the cells were overgrown, they were used for subsequent experiments. For the construction of the differentiation model of CPMs ( Cai et al, 2018 ; Li et al, 2022 ), the cells were collected by observing the cell fusion degree at 50 and 100% (labeled as G1 and G2). In addition, when the cells were confluent to 90 to 95%, the differentiation medium containing 2% horse serum (Vivacell, Shanghai, China)14 and 1% penicillin/streptomycin (Solarbio, Beijing, China) was used for further culture.…”