CircTCF4 Suppresses Proliferation and Differentiation of Goat Skeletal Muscle Satellite Cells Independent from AGO2 Binding

Abstract

The proliferation and differentiation of mammalian skeletal muscle satellite cells (MuSCs) are highly complicated. Apart from the regulatory signaling cascade driven by the protein-coding genes, non-coding RNAs such as microRNAs (miRNA) and circular RNAs (circRNAs) play essential roles in this biological process. However, circRNA functions in MuSCs proliferation and differentiation remain largely to be elucidated. Here, we screened for an exonic circTCF4 based on our previous RNA-Seq data, specifically expressed during the development of the longest dorsal muscle in goats. Subsequently, the circular structure and whole sequence of circTCF4 were verified using Sanger sequencing. Besides, circTCF4 was spatiotemporally expressed in multiple tissues from goats but strikingly enriched in muscles. Furthermore, circTCF4 suppressed MuSCs proliferation and differentiation, independent of AGO2 binding. Finally, we conducted Poly(A) RNA-Seq using cells treated with small interfering RNA targeting circTCF4 and found that circTCF4 would affect multiple signaling pathways, including the insulin signaling pathway and AMPK signaling pathway related to muscle differentiation. Our results provide additional solid evidence for circRNA regulating skeletal muscle formation.

METTL3 Promotes the Differentiation of Goat Skeletal Muscle Satellite Cells by Regulating MEF2C mRNA Stability in a m6A-Dependent Manner

“…The Jianzhou big-eared goat is a breed of meat sheep bred by crossing the American Nubian goat with the native goat in Sichuan province, which is the second meat goat species after the Nanjiang Huang goat in China [ 60 ]. A standard diet (forage to concentrate ratio, 65:35) was provided to all healthy ewes (2–3 years old) twice daily, and they received free access to water in their stalls, followed by simultaneous estrus and artificial insemination treatments [ 61 ]. Goat fetuses and kids (females, n = 3) at 90 embryonic days (E90), 105 days (E105), 120 days (E120), 135 days (E135), and 3 days after birth (B3) were randomly selected (not related) and humanely sacrificed.…”

HuR Promotes the Differentiation of Goat Skeletal Muscle Satellite Cells by Regulating Myomaker mRNA Stability

“…Primary MuSCs were isolated and cultured from the longissimus dorsi (LD) muscle of a fetal goat (Chengdu Ma goat, female), as previously described [ 10 , 38 ]. MuSCs were seeded in 6-well (~2 × 10 5 cells per well) or 12-well (~1 × 10 5 cells per well) plates and culture in a growth medium (GM) consisting of Dulbecco’s Modified Eagle Medium (DMEM/high glucose, Meilunbio, Dalian, China) supplemented with 15% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA).…”

The emerging roles of circRNAs in traits associated with livestock breeding

“…The circTCF4 is expressed at a level that is highest in the gastrocnemius, followed by skeletal muscle, and at a level that is lowest in the liver. It suppresses the proliferation and differentiation of goat skeletal muscle (Zheng et al, 2022).…”