CircUBE2Q2 promotes differentiation of cattle muscle stem cells and is a potential regulatory molecule of skeletal muscle development

Abstract

Background The growth and development of muscle stem cells (MuSCs) are significant events known to affect muscle plasticity, disease, meat production, and meat quality, which involves the types and functions of mRNA and non-coding RNA. Here, MuSCs were cultured from Guangxi fetal cattle. RNA sequencing was used to analyze the RNA expression of mRNA and non-coding RNAs during the cell proliferation and differentiation phases. Results Two thousand one hundred forty-eight mRNAs and 888 non-coding RNAs were differentially expressed between cell proliferation and differentiation phases, including 113 miRNAs, 662 lncRNAs, and 113 circRNAs. RT-qPCR verified the differential expression levels of mRNAs and non-coding RNAs, and the differentially expressed circUBE2Q2 was subsequently characterized. Expression profile analysis revealed that circUBE2Q2 was abundant in muscle tissues and intramuscular fat. The expression of cricUBE2Q2 was also significantly upregulated during MuSCs myogenic differentiation and SVFs adipogenic differentiation and decreased with age in cattle muscle tissue. Finally, the molecular mechanism of circUBE2Q2 regulating MuSCs function that affects skeletal muscle development was investigated. The results showed that circUBE2Q2 could serve as a sponge for miR-133a, significantly promoting differentiation and apoptosis of cultured MuSCs, and inhibiting proliferation of MuSCs. Conclusions CircUBE2Q2 is associated with muscle growth and development and induces MuSCs myogenic differentiation through sponging miR-133a. This study will provide new clues for the mechanisms by which mRNAs and non-coding RNAs regulate skeletal muscle growth and development, affecting muscle quality and diseases.

HDAC11 Regulates the Proliferation of Bovine Muscle Stem Cells through the Notch Signaling Pathway and Inhibits Muscle Regeneration

“…GOand the KEGG pathway were analyzed using the websites http://www.geneontology.org and http:// www.kegg.jp, respectively, as described previously. 22 Quantitative Reverse Transcription Polymerase Chain Reaction (qPCR). Total RNA was extracted using TRizol reagent (Invitrogen, Carlsbad, CA, U.S.A.) according to the instructions of the manufacturer.…”

“…MuSCs were cultured to 60−70% confluence, after which these cells were trypsinized, either plated onto 6-well plates at 5 × 10 5 cells/well or 96-well plates at 1 × 10 4 cells/well (Thermo Fisher Scientific), and incubated as described previously. 22 They were then transfected with p-HDAC11 and pCDNA3.1-eGFP, which were referred to as the overexpression and pcD3.1 (control) groups, respectively. The transfections were carried out using X-treme GENE HP DNA transfection reagent (Roche).…”

“…These were used with reference to previous reports. 22 Finally, protein bands were measured after treatment with Super Signal West Femto reagent purchased from Thermo Scientific (Waltham, MA, U.S.A.).…”

“…This test was also completed with reference to previous reports. 22 Dual-Luciferase Reporter (DLR) System Assay. Wild-type luciferase reporter plasmids of MyoD1 promoter (pGL3-MyoD1), pGL3 promoter (pGL3-P), and HDAC11 expression plasmids were transfected into 293T cells to analyze their targetting relationships.…”

“…This test was also completed with reference to previous reports. 22,23 Co-immunoprecipitation (co-IP) Assay. For each co-IP assay, proteins were extracted from MuSCs and grown in 5 × 10 cm dishes until fully confluent, according to the protocol of the manufacturer (Invent Biotechnologies, Plymouth, MN, U.S.A.).…”