Chronic manganese (Mn) overexposure causes a neurological disorder, referred to as manganism, exhibiting symptoms similar to parkinsonism. Dysfunction of the repressor element‐1 silencing transcription factor (REST) is associated with various neurodegenerative diseases such as Parkinson's disease, Alzheimer's disease, and Mn‐induced neurotoxicity, but its cellular and molecular mechanisms have yet to be fully characterized. Although neuronal REST is known to be neuroprotective, the role of astrocytic REST in neuroprotection remains to be established. We investigated if astrocytic REST in the striatal region of the mouse brain where Mn preferentially accumulates plays a role in Mn‐induced neurotoxicity. Striatal astrocytic REST was deleted by infusion of adeno‐associated viral vectors containing sequences of the glial fibrillary acidic protein promoter‐driven Cre recombinase into the striatum of RESTflox/flox mice for 3 weeks, followed by Mn exposure (30 mg/kg, daily, intranasally) for another 3 weeks. Striatal astrocytic REST deletion exacerbated Mn‐induced impairment of locomotor activity and cognitive function with further decrease in Mn‐reduced protein levels of tyrosine hydroxylase and glutamate transporter 1 (GLT‐1) in the striatum. Astrocytic REST deletion also exacerbated the Mn‐induced proinflammatory mediator COX‐2, as well as cytokines such as TNF‐α, IL‐1β, and IL‐6, in the striatum. Mn‐induced detrimental astrocytic products such as proinflammatory cytokines on neuronal toxicity were attenuated by astrocytic REST overexpression, but exacerbated by REST inhibition in an in vitro model using primary human astrocytes and Lund human mesencephalic (LUHMES) neuronal culture. These findings indicate that astrocytic REST plays a critical role against Mn‐induced neurotoxicity by modulating astrocytic proinflammatory factors and GLT‐1.