Abstract
Intramuscular fat (IMF) deposition is a complicated process, and most of the underlying regulators of this biological process are unknown. Here, we cloned the intact CDS of KLF4 gene, investigated the role of KLF4 by gaining or losing function in vitro and further explored the pathways of KLF4 regulating differentiation of intramuscular preadipocytes in goat. Our results show that goat KLF4 gene consists of 1,536 bp encoding a protein of 486 amino acids. The expression of KLF4 is higher in the lung while lower in the heart and muscle in goat. Knockdown of KLF4 mediated by siRNA technique significantly promotes intramuscular preadipocyte lipid accumulation and upregulates mRNA expression of adipogenic related genes including C/EBPα, C/EBPβ, and PPARγ in vivo cultured cells. Consistently, overexpression of KLF4 inhibits intramuscular adipocyte lipid accumulation and significantly downregulation gene expression of C/EBPβ, PPARγ, aP2, and Pref-1. Further, we found that other members of KLFs were upregulated or downregulated after interference or overexpression of KLF4, including KLF2 and KLF5–7. We also found that C/EBPβ was a potential target of KLF4, because it had an opposite expression pattern with KLF4 during the differentiation of intramuscular preadipocytes and had putative binding sites of KLF4. The dual-luciferase reporter assay indicated that overexpression of KLF4 inhibited the transcriptional activity of C/EBPβ. These results demonstrate that KLF4 inhibits the differentiation of intramuscular preadipocytes in goat by targeting C/EBPβ.
Highlights
Intramuscular fat (IMF) content is a crucial indicator for meat sensory quality evaluation and has been positively correlated with meat color, marbling, moisture content, and flavor (Cheng et al, 2015)
IMF deposition is a complicated process that is controlled by triacylglycerol catabolic or synthesis genes and a complex network of transcription factors, including CCAAT enhancer binding protein family, peroxisome proliferator-activated receptor gamma (PPARγ), KLF4 Inhibits Goat Preadipocytes Differentiation sterol regulatory element binding protein isoform 1c (SREBP1c), Krüppel-like factor (KLF) family, lipoprotein lipase (LPL), and fatty acid binding protein (FABP4) (Lowe et al, 2011)
Our results show that KLF4 inhibits the adipogenic differentiation of goat intramuscular preadipocytes through targeting C/EBPβ directly
Summary
Intramuscular fat (IMF) content is a crucial indicator for meat sensory quality evaluation and has been positively correlated with meat color, marbling, moisture content, and flavor (Cheng et al, 2015). Optimizing the number of intramuscular adipocytes can enhance IMF content. The balance between triglyceride (TG) anabolism and catabolism determines the volume of lipid droplets in intramuscular preadipocytes, which misguidedly cause IMF deposition (Gao and Zhao, 2009). IMF deposition is a complicated process that is controlled by triacylglycerol catabolic or synthesis genes and a complex network of transcription factors, including CCAAT enhancer binding protein family, peroxisome proliferator-activated receptor gamma (PPARγ), KLF4 Inhibits Goat Preadipocytes Differentiation sterol regulatory element binding protein isoform 1c (SREBP1c), Krüppel-like factor (KLF) family, lipoprotein lipase (LPL), and fatty acid binding protein (FABP4) (Lowe et al, 2011). It is believed that a complicated regulatory network exists in these molecular regulators and that many genes were involved in this biological process (Tontonoz et al, 1994; Tzameli et al, 2004). Little information is known about the molecular regulatory mechanisms understanding the IMF deposition
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