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Regulation by Cholesterol of the Gene Expression of Spot 14 in Adipocytes

Thyroid hormone responsive Spot 14 (THRSP) is a transcription factor localized to the liver and adipocytes. In addition to sensing triiodo‐L‐thyronine (T3) and glucose levels, previous studies have indicated the expression of Spot 14 is regulated by liver X receptor (LXR), a transcription factor activated by cholesterol. The known target genes of Spot 14 are specific to lipogenesis, suggesting it as a biochemical control point in the accumulation of lipids. In adipocytes, however, the regulation and function of Spot 14 have yet to be investigated. In this study, 3T3 L1 mouse adipocytes were depleted of cholesterol and/or supplemented with an LXR agonist or T3 in order to examine the regulation of Spot 14. Gene expression of these treated cells was measured through quantitative PCR for Spot 14 and its hypothesized target genes. In what appears to be through independent mechanisms, the expression of Spot 14 increased with both the additions of T3 and the LXR agonist, whereas the cholesterol depletion had the opposite effect showing a decrease in the mRNA. The proposed target genes fatty acid synthase (FAS), ATP‐citrate lyase (Acly), and phosphoenolpyruvate carboxykinase (Pck1), all enzymes involved in the synthesis of fatty acids, demonstrated a similar, yet T3 dominant response. In a related study perilipin, a protein found on the surface of large lipid droplets, was also analyzed. The expression of this gene exhibited a cholesterol dependent response similar to that of Spot 14. The results from this study provide further indication of the cholesterol‐mediated role of Spot 14 in the storage of lipids.