Hypothalamic circuitry underlying stress-induced insomnia and peripheral immunosuppression

The neural substrates of insomnia/hyperarousal induced by stress remain unknown. Here, we show that restraint stress leads to hyperarousal associated with strong activation of corticotropin-releasing hormone neurons in the paraventricular nucleus of hypothalamus (CRH^PVN) and hypocretin neurons in the lateral hypothalamus (Hcrt^LH). CRH^PVN neurons directly innervate Hcrt^LH neurons, and optogenetic stimulation of LH-projecting CRH^PVN neurons elicits hyperarousal. CRISPR-Cas9-mediated knockdown of the crh gene in CRH^PVN neurons abolishes hyperarousal induced by stimulating LH-projecting CRH^PVN neurons. Genetic ablation of Hcrt neurons or crh gene knockdown significantly counteracts restraint stress-induced hyperarousal. Single-cell mass cytometry by time of flight (CyTOF) revealed extensive changes to immune cell distribution and functional responses in peripheral blood during hyperarousal upon optogenetic stimulation of CRH^PVN neurons simulating stress-induced insomnia. Our findings suggest both central and peripheral systems are synergistically engaged in the response to stress via CRH^PVN circuitry.