Subplate neurons are among the earliest generated neurons in the cerebral cortex that orchestrate the development of the intra and extracortical connections. A large proportion of these neurons undergoes cell death throughout postnatal development. The remaining cells form the thin and compact cortical layer 6b (L6b) in murine cortex. The functional aspects of L6b have received little attention compared to the other layers. Recent studies have argued that the extensive intracortical connections and cortico-thalamic-cortical loops of L6b neurons might enable sensory processing to extract salient features of sensory stimuli (Zolnik et al., 2023). This layer is characterized by being the only cortical layer containing projection neurons that respond directly to the polypeptide Orexin, a neuropeptide known to be involved in cortical arousal and emotive behaviour (Bayer et al., 2002). Nevertheless, the exact mechanisms underlying these responses are not fully understood. We hypothesize that cortical circuits activated by these orexin-sensitive L6b neurons (Tg(Drd1a-cre)FK164Gsat/Mmucd) are not only responsible for detecting salient features of sensory stimuli but are also involved in the regulation of emotional states. Here, we show the Drd1a-Cre+ neurons are selectively sensitive to orexin and gate the activation of the prefrontal network. We also found that chronic inactivation of these neurons abolishes the orexinergic activation of the prefrontal cortex, and explains the anxiolytic effects of chronically silencing this subpopulation (Drd1a-Cre+:Snap25fl/fl) across the cortical mantle on several anxiety-related behavioural paradigms. This suggests that the orexin-responsive cortical layer 6b neuronal population is a potential target for the manipulation of anxiety-like behaviour.