Abstract
Adipocytes are derived from pluripotent mesenchymal stem cells through adipogenesis. Pre-adipocyte differentiation in poultry greatly influences fat deposition and meat quality. Circular RNAs (circRNAs) have an important function in cancer and some differentiation processes. Herein, high-throughput transcriptome sequencing was used to detect circRNAs present in cherry valley duck pre-adipocyte and adipocyte differentiation over 3 days. We identified 9,311 circRNAs and 141 differentially expressed circRNAs. Sequencing results were verified through qRT-PCR using seven randomly selected circRNAs, and competing endogenous RNA (ceRNA) networks were exhibited by ten important circRNAs in duck adipocyte differentiation. circRNA plexin A1 (circ-PLXNA1) was detected in duck adipocytes and mainly expressed in adipose, leg muscle and liver. Inhibition of circ-PLXNA1 limited the differentiation of duck adipocyte. There were four corresponding miRNAs for circ-PLXNA1 and 313 target genes for those miRNAs. CeRNA“circ-PLXNA1/miR-214/CTNNB1 axis” was focused and verified by a dual-luciferase reporter experiment. After co-transfection of cells with si-circ-PLXNA1 and miR-214 mimics, the expression level of CTNNB1 was down-regulated, triglyceride content and the adipogenic capacity of preadipocytes decreased. While there were no significant change after si-CTNNB1 transfection. All these results provide further insight into the circRNAs, especially for circ-PLXNA1 in duck adipocyte differentiation.
Highlights
The intramuscular fat content of livestock has received much attention due to its high contribution to meat quality
All of the marker genes, including Delta-like non-canonical Notch ligand 1 (DLK1), LPL, CCAAT/enhancer-binding protein-α (C/EBPα), peroxisome proliferator-activated receptor γ (PPARγ), fatty-acid binding protein 4 (FABP4), and fatty acid synthetase (FAS) were detected in the culture of adipocyte cells (Fig 1F)
The expression levels of C/EBPα, PPARγ, FABP4, and FAS were increased and DLK1 was decreased in differentiated adipocytes compared to the pre-adipocytes, which showed normal differentiation progress
Summary
The intramuscular fat content of livestock has received much attention due to its high contribution to meat quality. Understanding or manipulating the process may, lead to better meat quality. Fat deposition is influenced by both the number of fat cells and the enlargement of adipocytes in muscle. The proliferation and differentiation of pre-adipocytes lead to the formation of adipose tissue, and pre-adipocyte differentiation is characterized by a coordinated increase in gene expression and lipid droplet accumulation [1,2,3].
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