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. 2023 Nov 17;14(11):2088.
doi: 10.3390/genes14112088.

FOS Inhibits the Differentiation of Intramuscular Adipocytes in Goats

Tingting Hu  1   2   3 , Zhibin Li  1   2   3 , Chengsi Gong  1   2   3 , Yan Xiong  1   2   3 , Shiyu Sun  1   2 , Jiani Xing  1   2   3 , Yanyan Li  1   2   3 , Ruiwen Li  4 , Youli Wang  1   2   3 , Yong Wang  1   2   3 , Yaqiu Lin  1   2   3
Affiliations

FOS Inhibits the Differentiation of Intramuscular Adipocytes in Goats

Tingting Hu et al. Genes (Basel). .

Abstract

Goat intramuscular fat (IMF) deposition is precisely regulated by many key genes as well as transcription factors. Nevertheless, the potential of the regulators of goat IMF deposition remains undefined. In this work, we reported that the transcription factor FOS is expressed at a low level at the early differentiation stage and at a high level in late differentiation. The overexpression of FOS inhibited intramuscular adipocyte lipid accumulation and significantly downregulated the expressions of PPARγ, C/EBPβ, C/EBPα, AP2, SREBP1, FASN, ACC, HSL, and ATGL. Consistently, the knockdown of FOS, facilitated by two distinct siRNAs, significantly promoted intramuscular adipocyte lipid accumulation. Moreover, our analysis revealed multiple potential binding sites for FOS on the promoters of PPARγ, C/EBPβ, and C/EBPα. The expression changes in PPARγ, C/EBPβ, and C/EBPα during intramuscular adipogenesis were opposite to that of FOS. In summary, FOS inhibits intramuscular lipogenesis in goats and potentially negatively regulates the expressions of PPARγ, C/EBPβ, and C/EBPα genes. Our research will provide valuable data for the underlying molecular mechanism of the FOS regulation network of intramuscular lipogenesis.

Keywords: FOS; cell differentiation; goat; intramuscular adipocytes.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Subcellular localization of FOS and expression patterns during differentiation of goat intramuscular adipocytes. (A) The upper panels show the localization of PEGFP-N1, and the bottom panels show the localization of the FOS protein in intramuscular adipocytes, as indicated by the white arrow in the figure. The scale bar represents 50 µm. (B) The expression changes in FOS during 0–5 days of induced differentiation of goat intramuscular adipocytes (n = 6). “**” means an extremely significant difference (P < 0.01).
Figure 2
Figure 2
Overexpression of FOS inhibits goat intramuscular adipocyte differentiation. (A) Overexpression efficiency of FOS detected using qPCR (n = 6). (B,C) Oil Red O staining (×200) (n = 3), scale bar represents 50 µm, and the value of OD at 490 nm. (D) Bodipy and DAPI stainings (×200); scale bar represents 50 µm. The green fluorescence represents lipid droplets. “**” means an extremely significant difference (P < 0.01).
Figure 3
Figure 3
Overexpression of FOS suppresses the expression of adipogenesis genes. (A–C) The expressions of AP2, PPARγ, C/EBPβ, C/EBPα, SREBP1, FASN, ACC, SCD1, HSL, ATGL, and LPL in goat intramuscular adipocytes in NC and OE groups. (D) The cellular TG levels in goat intramuscular adipocytes in negative control (NC) and overexpression of FOS treatment groups. (E) Protein expressions of PPARγ and C/EBPβ. “*” means a significant difference (P < 0.05), “**” means an extremely significant difference (P < 0.01) and “ns” means no difference.
Figure 4
Figure 4
Knockdown of FOS increases accumulation of lipid droplets in goat intramuscular adipocytes. (A) The knockdown efficiency of the FOS detection. (B,C) Oil Red O staining (×200), scale bar represents 50 µm, and the value of OD at 490 nm. (D) Bodipy and DAPI stainings (×200); scale bar represents 50 µm. The green fluorescence represents lipid droplets. “**” means an extremely significant difference (P < 0.01).
Figure 5
Figure 5
Knockdown of FOS upregulates positive adipocyte differentiation genes and downregulates lipid metabolism-related genes. (A–C) The expressions of lipogenic-related genes in the NC and knockdown FOS groups. (D) Detection of cellular TG contents in the NC and FOS knockdown groups. (E) PPARγ and C/EBPβ protein expression levels in NC and knockdown FOS groups. “*” means a significant difference (P < 0.05), “**” means an extremely significant difference (P < 0.01) and “***” means an extremely significant difference (P < 0.001).
Figure 6
Figure 6
FOS exhibits an expression pattern opposite to those of PPARγ, C/EBPβ, and C/EBPα during adipocyte differentiation. (A) the PPARγ expression trend during intramuscular adipogenesis (n = 6). (B) The C/EBPβ expression trend during intramuscular adipogenesis. (C) The C/EBPα expression trend during intramuscular adipogenesis. (D) FOS binding sites on the promoters of PPARγ and C/EBPβ. Blue circles represent the FOS binding sites.
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