Abstract
Suppressor of cytokine signaling-3 (SOCS3) is a negative regulator of actions of cytokines and the metabolic hormone leptin. In the hypothalamus, SOCS3 is induced in response to several conditions such as inflammation and high-fat diet feeding, modulates cellular signaling of cytokines and leptin, and mediates the effects of these biological conditions. However, signaling mechanisms controlling hypothalamic Socs3 expression remains to be fully established. To facilitate the identification of molecular pathways of Socs3 induction, we generated a real-time gene expression reporter mouse of Socs3 (Socs3-Luc mice). We successfully detected a remarkable increase in luciferase activity in various tissues of Socs3-Luc mice in response to a peripheral injection of lipopolysaccharide, a potent inducer of inflammation, reflecting expression levels of endogenous Socs3 mRNA. Using ex vivo hypothalamic explants of Socs3-Luc mice, we demonstrate that hypothalamic luciferase activity was significantly elevated in slices stimulated with known inducers of Socs3 such as proinflammatory cytokines IL-6, IL-1β, and TNF-α, lipopolysaccharide, and cAMP-inducing agent forskolin. Using the ex vivo model, we found glycogen synthase kinase-3 (GSK3)β-specific inhibitors to be potent inducers of Socs3. Furthermore, pharmacological inhibitors of β-catenin, a downstream mediator of GSK3β signaling, reduced Socs3 luciferase activity ex vivo. Finally, hypothalamic inhibition of GSK3β hindered leptin-induced phosphorylation of signal transducers and activators of transcription 3 in hypothalamic explants. These results suggest that the Socs3-luciferase mouse is useful for in vivo monitoring of Socs3 gene expression and for ex vivo slice-based screening to identify signaling pathways that control Socs3 in the hypothalamus.