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To examine whether morphine induces constipation in mice by inhibiting PVN Glu neuronal activity via MOR, we performed immunofluorescence staining in RV-tracing brain slices of normal mice revealed that EGFP + /MOR + co-labelled cells across several regions, including the PVN, parabrachial pigmented nucleus (PBP), inferior colliculus (IC), primary somatosensory cortex (S1), periaqueductal gray (PAG), locus coeruleus (LC), mesencephalic reticular formation, raphe magnus nucleus, retrorubral field, parasubthalamic nucleus, and dorsal raphe nucleus (Fig