Zfp423 promotes adipogenic differentiation of bovine stromal vascular cells

Yan Huang; Arun Kr Das; Qi-Yuan Yang; Mei-Jun Zhu; Min Du

doi:10.1371/journal.pone.0047496

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Zfp423 promotes adipogenic differentiation of bovine stromal vascular cells

Journal article

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Peer reviewed

Zfp423 promotes adipogenic differentiation of bovine stromal vascular cells

Yan Huang, Arun Kr Das, Qi-Yuan Yang, Mei-Jun Zhu and Min Du

PloS one, Vol.7(10), pp.e47496-e47496

2012

DOI: https://doi.org/10.1371/journal.pone.0047496

Handle:

https://hdl.handle.net/2376/111373

PMCID: PMC3468566

PMID: 23071815

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Abstract

CCAAT-Enhancer-Binding Protein-alpha - metabolism

Receptor, Platelet-Derived Growth Factor alpha - metabolism

Linear Models

DNA Primers - genetics

Transcription Factors - genetics

DNA-Binding Proteins - genetics

Muscle, Skeletal - cytology

PPAR gamma - metabolism

Blotting, Western

Adipogenesis - physiology

Meat

DNA-Binding Proteins - metabolism

Transcription Factors - metabolism

Cattle

Azo Compounds

Plasmids - genetics

Mice

Stromal Cells - physiology

Cell Differentiation - physiology

Real-Time Polymerase Chain Reaction

Transforming Growth Factor beta - metabolism

Transfection

Intramuscular fat or marbling is critical for the palatability of beef. In mice, very recent studies show that adipocytes and fibroblasts share a common pool of progenitor cells, with Zinc finger protein 423 (Zfp423) as a key initiator of adipogenic differentiation. To evaluate the role of Zfp423 in intramuscular adipogenesis and marbling in beef cattle, we sampled beef muscle for separation of stromal vascular cells. These cells were immortalized with pCI neo-hEST2 and individual clones were selected by G418. A total of 288 clones (3×96 well plates) were isolated and induced to adipogenesis. The presence of adipocytes was assessed by Oil-Red-O staining. Three clones with high and low adipogenic potential respectively were selected for further analyses. In addition, fibro/adipogenic progenitor cells were selected using a surface marker, platelet derived growth factor receptor (PDGFR) α. The expression of Zfp423 was much higher (307.4±61.9%, P<0.05) in high adipogenic cells, while transforming growth factor (TGF)-β was higher (156.1±48.7%, P<0.05) in low adipogenic cells. Following adipogenic differentiation, the expression of peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer binding protein α (C/EBPα) were much higher (239.4±84.1% and 310.7±138.4%, respectively, P<0.05) in high adipogenic cells. Over-expression of Zfp423 in stromal vascular cells and cloned low adipogenic cells dramatically increased their adipogenic differentiation, accompanied with the inhibition of TGF-β expression. Zfp423 knockdown by shRNA in high adipogenic cells largely prevented their adipogenic differentiation. The differential regulation of Zfp423 and TGF-β between low and high adipogenic cells is associated with the DNA methylation in their promoters. In conclusion, data show that Zfp423 is a critical regulator of adipogenesis in stromal vascular cells of bovine muscle, and Zfp423 may provide a molecular target for enhancing intramuscular adipogenesis and marbling in beef cattle.

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Title: Zfp423 promotes adipogenic differentiation of bovine stromal vascular cells
Creators: Yan Huang - Developmental Biology Group, Department of Animal Science, University of Wyoming, Laramie, WY, USA
Arun Kr Das
Qi-Yuan Yang
Mei-Jun Zhu
Min Du
Publication Details: PloS one, Vol.7(10), pp.e47496-e47496
Academic Unit: Department of Animal Sciences
Publisher: United States
Grant note: R01 HD067449 / NICHD NIH HHS R01HD067449 / NICHD NIH HHS
Identifiers: 99900547613701842
Language: English
Resource Type: Journal article