Global Analysis of Gene Expression Mediated By Orexin Receptor 1 Signaling in a Hypothalamic Cell Line. Mus musculus

The orexins/hypocretins and their cognate G-protein coupled receptors have been widely studied due to their associations with various behaviors and cellular processes. However, the detailed downstream signaling cascades that mediate these effects are not completely understood. We report the generation of a neuronal model cell line that stably expresses orexin receptor 1 and an RNA-Seq analysis of changes in gene expression seen upon receptor activation. Upon treatment with orexin, several related families of related transcription factors are transcriptionally regulated, including the early growth response genes (Egr), the Kruppel-like factors (Klf), and the Nr4a subgroup of nuclear hormone receptors. Furthermore, the transcriptional effects observed showed a degree of similarity with data from in vivo sleep deprivation microarray studies, supporting the physiological relevance of the data set. These results provide new insight into the molecular signaling events that occur during orexin receptor 1 signaling and support a role for orexin signaling in the stimulation of wakefulness during sleep deprivation studies. Overall design: Differential gene expression was evaluated via RNA-Seq upon activation of OxR1 for 3 or 8 hours, compared to vehicle treated cells. All samples were done in triplicate.