miR-214-5p Regulating Differentiation of Intramuscular Preadipocytes in Goats via Targeting KLF12

Abstract

Intramuscular fat (i.m.) is an adipose tissue that is deposited between muscle bundles. An important type of post-transcriptional regulatory factor, miRNAs, has been observed as an important regulator that can regulate gene expression and cell differentiation through specific binding with target genes, which is the pivotal way determining intramuscular fat deposition. Thus, this study intends to use RT-PCR, cell culture, liposome transfection, real-time fluorescent quantitative PCR (qPCR), dual luciferase reporter systems, and other biological methods clarifying the possible mechanisms on goat intramuscular preadipocyte differentiation that is regulated by miR-214-5p. Ultimately, our results showed that the expression level of miR-214-5p peaked at 48 h after the goat intramuscular preadipocytes were induced for adipogenesis. Furthermore, after inhibition of the expression of miR-214-5p, the accumulation of lipid droplets and adipocyte differentiation in goat intramuscular adipocytes were promoted by the way of up-regulation of the expression level of lipoprotein lipase (LPL) (p < 0.05) and peroxisome proliferator-activated receptor gamma (PPARγ) (p < 0.01) but inhibited the expression of hormone-sensitive lipase (HSL) (p < 0.01). Subsequently, our study confirmed that Krüppel-like factor 12 (KLF12) was the target gene of miR-214-5p. Inhibition of the expression of KLF12 promoted adipocyte differentiation and lipid accumulation by upregulation of the expression of LPL and CCAAT/enhancer binding protein (C/EBPα) (p < 0.01). Overall, these results indicated that miR-214-5p and its target gene KLF12 were negative regulators in progression of goat preadipocyte differentiation. Our research results provided an experimental basis for finally revealing the mechanism of miR-214-5p in adipocytes.

Genome-wide analysis of circular RNA-mediated ceRNA regulation in porcine skeletal muscle development

“…For some of the miRNAs in this ceRNA network, a role in regulating proliferation and differentiation of skeletal muscle satellite cells has already been reported in other animals. These include miR-370 [66], miR-214 [67], miR-133 [68], and miR-885 [69]. We predicted that circ_0000576 could act as a sponge for ssc-miR-15a, ssc-miR-98, and ssc-miR-885-3p, modulating the target genes of these miRNAs to regulate muscle growth and development.…”

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 ].…”

miRNA expression patterns in blood leukocytes and milk somatic cells of goats infected with small ruminant lentivirus (SRLV)

“…In the studies of caprine and ovine biology, the subject of research in relation to several forms of RNA e.g. : miRNAs, circle RNAs and lncRNAs (long noncoding RNAs) are mainly those about animals pregnancy 26 – 29 , functioning of the tissues and organs 30 – 33 , mammary gland health state 34 , 35 , or the function of the milk extracellular vesicles 36 . However, currently, only very limited information exists on the participation of miRNAs in SRLV infection.…”