chi-miR-99b-3p Regulates the Proliferation of Goat Skeletal Muscle Satellite Cells In Vitro by Targeting Caspase-3 and NCOR1

Abstract

We previously found that chi-miR-99b-3p was highly expressed in the skeletal muscle of 7-month-old (rapid growth period) goats and speculated that it may be associated with muscle development. To further investigate the role of chi-miR-99b-3p in goats, we found that chi-miR-99b-3p acted as a myogenic miRNA in the regulation of skeletal muscle development. Dual-luciferase reporter assays, qRT-PCR, and Western blot results confirmed that Caspase-3 and nuclear receptor corepressor 1 were direct targets for chi-miR-99b-3p as their expression was inhibited by this miR. Cell proliferation and qRT-PCR assays showed that chi-miR-99b-3p promoted proliferation through relevant targets and intrinsic apoptosis-related genes in goat skeletal muscle satellite cells (SMSCs), whereas inhibition of chi-miR-99b-3p had the opposite effect. Furthermore, integrative transcriptomic analysis revealed that overexpression of chi-miR-99b-3p induced various differentially expressed (DE) genes mainly associated with the cell cycle, relaxin signaling pathway, DNA replication, and protein digestion and absorption. Notably, most of the cell-cycle-related genes were downregulated in SMSCs after miR-99b-3p upregulation, including the pro-apoptosis-related gene BCL2. In addition, 47 DE miRNAs (16 upregulated and 31 downregulated) were determined by Small RNA-sequencing in SMSCs after chi-miR-99b-3p overexpression. Based on the KEGG enrichment analysis, we found that these DE miRNAs were involved in the biological pathways associated with the DE genes. Our study demonstrated that chi-miR-99b-3p was an effective facilitator of goat SMSCs and provided new insights into the mechanisms by which miRNAs regulate skeletal muscle growth in goats.

LncRNA-004686 regulates muscle fibre type switching in chicken embryos by affecting the expression of MYH1B

“…Recent research has demonstrated that whereas MYOD, MRF4, MHC, and other genes primarily govern muscle cells fusion, MSTN, Myf5, and IGF-1 are primarily involved in the proliferation of muscle cell 27,28,29,30,31 . The number of muscle cells did not change considerably after transfection of pcDNA3.1 (+)-004686 into chicken primary myoblasts, while the expression of IGF-1, a marker gene for muscle cell proliferation, increased dramatically.…”

Temporal transcriptomic changes in microRNAs involved in the host immune response and metabolism during Neospora caninum infection

“…ovary [ 45 ], liver [ 46 ], pituitary [ 47 ], lung [ 48 ]) and types of cells (e.g. skeletal muscle satellite cells [ 49 ], skin fibroblast cells [ 50 ], Leydig cells [ 51 ], blood leukocytes and milk somatic cells [ 52 ], intramuscular preadipocytes [ 53 ], endometrial epithelium cells [ 54 ]) in physiological and pathological processes in goats, including in response to infectious diseases. For example, the expression of 316 DEmiRNAs was found in peripheral blood mononuclear cells of goats infected with peste des petits ruminants virus, and most of the predicted targets of these DEmiRNAs were found to be involved in immune escape [ 55 ].…”

The Function and Regulation Mechanism of Non-Coding RNAs in Muscle Development

“…miR-23a promotes myogenic differentiation of fetal bovine skeletal muscle-derived progenitor cells by targeting the MyoD family inhibitor structural domain-containing (MDFIC) genes [66]. miR-2425-5p targets the 3 -UTR of RAD9 homolog A (RAD9A) and myogenesis (MYOG) to promote proliferation and inhibit differentiation [67] in sheep and goats, including miR-181a, miR-29a, miR-99b-3p, and miR-27a-3p [69][70][71][72] and in rabbits and pigs, including miR-194-5p and miR-22, respectively [80,81]. The specific mechanisms of action of these miRNAs will not be elaborated.…”