Abstract
Adipose development is regulated by a series of complex processes, and non-coding RNAs (ncRNAs), including circular RNAs (circRNAs), play important roles in regulating proliferation and differentiation of adipocytes. In this study, we profiled circRNA expression in cattle fat tissue during calf and adult developmental stages and detected 14,274 circRNA candidates. Some circRNAs are differentially expressed between two developmental stages. We characterized circFUT10, named for its host gene FUT10, a highly expressed and abundant circRNA. Luciferase screening, an RNA-binding protein immunoprecipitation (RIP) assay, quantitative real-time PCR, and western blotting assays indicated that circFUT10 directly binds let-7c/let-e, and PPARGC1B (peroxisome proliferator-activated receptor γ coactivator 1-β) is identified as a target of let-7c. Flow cytometry, EdU (5-ethynyl-2′-deoxyuridine) incorporation, a CCK-8 (cell counting kit-8) assay, oil red O staining, and western blotting assays demonstrated that circFUT10 promotes adipocyte proliferation and inhibits cell differentiation by sponging let-7c. The results demonstrate that circFUT10 binding of let-7c promotes cell proliferation and inhibits cell differentiation by targeting PPARGC1B in cattle adipocytes.
Highlights
Adipogenesis is a complex and precisely orchestrated process mediated by a network of adipocyte regulatory factors
Tissue We identified RNAs in calf and adult cattle fat tissue using Ribo-Zero RNA sequencing (RNA-seq),[25] with 89–102 and 91–118 million unique mapped clean reads having been acquired from the calf and adult cattle libraries, respectively (Table 1)
We found that 64.4% of all of the reads mapped to the protein coding region, 13.9% of reads mapped to miscellaneous RNA, almost no reads mapped to pseudogene, small nucleolar RNA, or miRNA regions, and 21.2% of the reads were not mapped (Figure 1A)
Summary
Adipogenesis is a complex and precisely orchestrated process mediated by a network of adipocyte regulatory factors. The peroxisome proliferator-activated receptor g (PPARg) and CCAAT/enhancerbinding proteins (C/EBPs), a family of transcription factors composed of six members, named C/EBPa to C/EBPz, mediate transcription.[1,2] PPARg is induced during terminal differentiation and is required for the activation of a number of genes with adipogenic.[3] Three C/EBPs are involved in the differentiation of adipocytes C/EBPa, C/EBPb, and C/EBPd. C/EBPb and C/EBPd are expressed early in the induction of differentiation, and they initiate the transcription of PPARg and C/EBPa to control the differentiation of adipocytes.[4,5] the major molecular pathways of adipogenesis are understood, the regulatory scenario is far from complete. Recent studies have demonstrated roles of non-coding RNA (ncRNA) and circular RNA (circRNA) in the adipogenesis regulatory network.[6,7]
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