Lipin expression preceding peroxisome proliferator-activated receptor-γ is critical for adipogenesis in vivo and in vitro

We recently identified mutations in the lipin gene, Lpin1, as the cause of lipodystrophy in the fatty liver dystrophy (fld) mouse. Here we identify impaired adipocyte differentiation as the basis for lipodystrophy in lipin-deficient mice and demonstrate that lipin is required for normal induction of the adipogenic gene transcription program. We found that the reduced adiposity in chow fed fld mice and resistance to obesity in fld mice fed a high-fat diet is associated with reduced adipogenic gene expression. Using primary mouse embryonic fibroblasts isolated from fld mice, we confirmed that lipin deficiency prevents normal lipid accumulation and induction of key adipogenic genes, including peroxisome proliferator-activated receptor (PPAR)γ and CCAAT enhancer-binding protein (C/EBP)α. However, our previous studies of daily gene expression in differentiating 3T3-L1 preadipocytes indicated that lipin expression is undetectable until about day 3 of differentiation, at a point after PPARγ and C/EBPα gene expression is established. This paradox was resolved by examining gene expression at 10-h intervals during 3T3-L1 cell differentiation, leading to detection of transient lipin expression at 10 h into the differentiation program, prior to the induction of PPARγ and C/EBPα. Consistent with a requirement for lipin expression upstream of PPARγ, differentiation of lipin-deficient mouse embryonic fibroblasts could be rescued by ectopic expression of PPARγ. Thus, we conclude that lipin expression is required prior to PPARγ during adipocyte differentiation.