miR-10a-5p Inhibits the Differentiation of Goat Intramuscular Preadipocytes by Targeting KLF8 in Goats

Abstract

Intramuscular fat contributes to the improvement of meat quality of goats. MicroRNAs (miRNAs) have been reported to regulate adipocyte differentiation and maturation. The aim of our study was to clarify whether miR-10a-5p regulates goat intramuscular preadipocyte (GIPC) differentiation and its direct downstream signaling pathway. GIPCs were isolated from longissimus dorsi, whose miR-10a-5p level was measured at different time point of differentiation induction. Adipogenic differentiation of the GIPCs was evaluated by Oil Red O and BODIPY staining, and the expression changes of adipogenic genes like ACC, ATGL, CEBPβ, PPARγ, etc. Related mechanisms were verified by qPCR, a bioinformatic analysis, a dual-luciferase reporter assay, overexpression, and siRNA transfection. Oil Red O and BODIPY staining both with adipogenic gene detection showed that miR-10a-5p suppressed the accumulation of lipid droplets in GIPCs and inhibited its differentiation. The dual-luciferase reporter assay experiment revealed that miR-10a-5p regulates GIPC differentiation by directly binding to KLF8 3’UTR to regulate its expression. Thus, the results indicated that miR-10a-5p inhibits GIPC differentiation by targeting KLF8 and supply a new target for fat deposition and meat quality improvement.

miR-130b duplex (miR-130b-3p/miR-130b-5p) negatively regulates goat intramuscular preadipocyte lipid droplets accumulation by inhibiting Krüppel-like factor 3 expression

“…The same result was observed in the rescue experiment of this study. Also, the same problem has been found in previous studies ( Guo et al, 2020 ; Xu et al, 2021 ; Chen et al, 2022 ). In addition, pref-1 , a well-known negative regulator of adipogenesis, was positively correlated with miR130b duplex (miR-130b-3p and miR-130b-5p) and KLF3 regulated goat intramuscular adipocyte differentiation.…”

“…The medium was changed 24 h later. The preadipocytes were then identified by Oil red O staining at different time points (3, 24, 48, 72, 96, and 120 h) with differentiation induction ( Xu et al, 2021 ). With the extension of differentiation induction, lipid droplets became larger and accumulation increased.…”

“…After transfection of the corresponding fragments into goat intramuscular preadipocytes and inducing differentiation for 48 h, Oil red O and Bodipy were used to detect the formation of lipid droplets, and procedures for staining and quantitative analysis have been provided in our previous study ( Xu et al, 2021 ). Briefly, the cells were washed twice with PBS and fixed with 4% paraformaldehyde at room temperature for 15 min.…”

miR-10a-5p Regulates the Proliferation and Differentiation of Porcine Preadipocytes Targeting the KLF11 Gene

“…In this study, miR-10a-5p enhanced adipogenic differentiation in porcine precursor adipocytes. However, within goat intramuscular preadipocytes, it inhibited differentiation by targeting KLF8 [38]. This discrepancy might be due to interspecies differences.…”

CircADAMTS16 Inhibits Differentiation and Promotes Proliferation of Bovine Adipocytes by Targeting miR-10167-3p

“…Recent studies have shown that ncRNAs, including microRNAs (miRNAs), long non-coding RNAs (lncRNAs) and circRNAs, are capable of participating in the process of lipid metabolism, including the proliferation and differentiation of adipocytes, the formation of fat droplets, and the activation of adipocyte-specific genes, as well as the regulation of genes during the development and remodeling of adipose tissue [ 8 , 9 , 10 ]. In goats, miR-10a-5p targets Krüppel-like factor 8 ( KLF8 ) and inhibits differentiation of intramuscular preadipocytes [ 11 ]. Through attenuation of phosphorylation of mitogen-activated protein kinase (MAPK)-p38 and MAPK-extracellular signal-regulated kinase 1/2 (ERK1/2), lnc-FR332443 positively regulates Runx1 expression and suppresses adipocyte differentiation in mouse adipocytes [ 12 ].…”