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Peroxisome proliferator-activated receptor- activation and long-chain fatty acids alter lipogenic gene networks in bovine mammary epithelial cells to various extents

A. K. G. Kadegowda^*, M. Bionaz^,, L. S. Piperova^*, R. A. Erdman^* and J. J. Loor^,,1

^* Department of Animal and Avian Sciences, University of Maryland, College Park 20742
Mammalian NutriPhysioGenomics, Department of Animal Sciences, and
Division of Nutritional Sciences, University of Illinois, Urbana 61801

¹ Corresponding author: jloor{at}illinois.edu

Several long-chain fatty acids (LCFA) are natural ligands of nonruminant peroxisome proliferator-activated receptor- (PPARG), which, along with its lipogenic target genes, is upregulated in bovine mammary tissue during lactation. Thus, PPARG might represent an important control point of bovine milk fat synthesis. We tested lipogenic gene network expression via quantitative PCR of 19 genes in bovine mammary epithelial cells cultured with 16:0, 18:0, cis-9 18:1, trans-10 18:1, trans-10,cis-12 18:2 [t10c12 conjugated linoleic acid (CLA)], 20:5, ethanol (control), and the PPARG agonist rosiglitazone (ROSI). Triplicate cultures were maintained for 12 h with 50 µM ROSI or 100 µM LCFA. Responses common to 16:0 and 18:0 relative to the control included significantly greater expression of INSIG1 (+298%, +92%), AGPAT6 (+137%, +169%), FABP3 (+755%, +338%), and FABP4 (+171%, 157%). These were coupled with greater intracellular lipid droplet formation and mRNA of ACSS2, LPIN1, SCD, and SREBF2 in response to 16:0, and greater DGAT1 and THRSP with 18:0. Trans-10 18:1 and t10c12 CLA reduced expression of FASN (–60%, –31%), SCD (–100%, –357%), and SREBF1 (–49%, –189%). Furthermore, t10c12 CLA downregulated ACSS2, FABP3, INSIG1, SREBF2, and THRSP expression. Expression of SREBF1 was lower with cis-9 18:1 (–140%) and 20:5 (–125%) compared with the control. This latter LCFA also decreased SCD, SREBF2, and LPL expression. No effects of LCFA or ROSI on PPARG were observed, but ROSI upregulated (+39% to +269%) expression of ACACA, FASN, LPIN1, AGPAT6, DGAT1, SREBF1, SREBF2, and INSIG1. Thus, these genes are putative PPARG target genes in bovine mammary cells. This is the first report showing a direct effect of trans-10 18:1 on bovine mammary cell lipogenic gene expression. The coordinated upregulation of lipogenic gene networks in response to ROSI and saturated LCFA offers support for PPARG activation in regulating bovine milk fat synthesis.

Key Words: nuclear receptor • milk fat • genomics

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Adipose tissue lipogenic gene networks due to lipid feeding and milk fat depression in lactating cows

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