The Dorsal Raphe Nucleus (DRN) is the largest of the serotonergic nuclei, but it also comprises a wide variety of neuronal subpopulations, including dopaminergic cells (DRNDA) and astrocytes, which may represent a source of local DRN modulation. The DRN participates in a wide range of behavioral responses related to the processing of aversive stimuli, leading to anxiety state. While the role of DRN5-HT and DRNDA in valence processing is emerging, the contribution of DRN astrocytes (DRNAstro), as well as the underpinning cellular mechanisms, remain elusive. Unpublished data from the lab indicates that the presentation of aversive stimuli triggers a reliable increase both in DRN-astrocytic Ca2+ activity and noradrenergic (NE) signal in the DRN. Furthermore, in freely moving mice, optogenetic stimulation of Locus Coeruleus (LC) projections to the DRN decreases speed and distance travelled, which might be an indicator of freezing responses. In this study, we aim to investigate how DRNAstro and regulation of their activity by LC-NE projections influence stimulus valence processing within the DRN circuitry. To this purpose, we combine intersectional viral strategies, ex-vivo two-photon Ca2+ imaging and in vivo fiber-photometry in a behavioral task based on the presentation of compound stimuli following Pavlovian learning. A similar behavioral conceptualization has been shown to reliably assess the behavioral impact of stimuli with uncertain outcome in multiple species, including humans. Thus, the results of our investigation could be instrumental in relating dysfunctional DRN modulation to the pathophysiology of comorbid affective symptoms in neurological disorders exhibiting varying degrees of astrocyte involvement.