The Bed Nucleus of the Stria Terminalis–Deep Mesencephalic Nucleus Circuit Linking Emotion and Wakefulness

Abstract

The bed nucleus of the stria terminalis (BNST), a part of the extended amygdala, integrates emotional and arousal-related signals. While GABAergic BNST (GABA BNST ) neurons have been implicated in promoting transitions from non-rapid eye movement (NREM) sleep to wakefulness, their downstream mechanisms remain unclear. Here, we identify a neuronal circuit through which GABA BNST neurons promote arousal via projections to a midbrain region known as the deep mesencephalic nucleus (DpMe), located within the broader mesencephalic reticular formation. In male mice, we used a combination of optogenetics, fiber photometry, neural ablation, and tracing approaches to dissect this circuit. Optogenetic stimulation of GABA BNST terminals in the DpMe during NREM sleep elicited rapid transitions to wakefulness and increased activity of glutamatergic DpMe (GLUT DpMe ) neurons, as assessed by c-fos mRNA expression and calcium imaging. Similarly, an aversive air-puff activated GLUT DpMe neurons, suggesting engagement by emotionally salient stimuli. Ablation of GLUT DpMe neurons markedly attenuated arousal responses triggered by GABA BNST stimulation, underscoring their essential role in this circuit. While monosynaptic rabies tracing revealed local input neurons to GLUT DpMe cells, in situ hybridization identified few Vgat- positive interneurons among them. These findings suggest that GABA BNST neurons may influence GLUT DpMe neurons through noncanonical GABAergic mechanisms or via more complex local circuits beyond a simple disinhibition model. Together, these findings delineate a previously uncharacterized BNST–DpMe circuit that allows emotionally relevant stimuli to override sleep and promote arousal. This pathway may contribute to stress-related sleep disturbances and represents a potential target for therapeutic treatments for sleep disorders associated with emotional dysregulation.