Stroke is a preeminent contributor of the global burden of diseases. Hypothermia, among other therapeutic approaches, has been proposed to promote neural survival after stroke, yet the physiological cooling remains ineffective and induces collateral side effects. The thermoregulatory mechanisms are under the control of neuronal identities that connect peripheral signals of the central nervous system, which are predominantly conveyed into the hypothalamus. Here, we aimed to identify a thermoregulatory neuronal subpopulation located at the dorsomedial hypothalamus (DMH) and investigate the DMH-projecting neurons that induce hypothermia against stroke. To investigate the thermoregulatory neuronal subpopulation, we inhibited DMHVgat2 neurons by delivering rAAV-EF1a-DIO-hM4D(Gi)-mCherry (n=10) or rAAV2/1-CAG-DIO-GFP (n=14) to DMH of Vgat2-cre mice. The mice were exposed to 30 minutes of middle cerebral artery occlusion followed by 72 h reperfusion. Then, animals were sacrificed and infarct volume, brain edema, neuronal survival, and DNA fragmentation were analyzed. Our findings revealed that hypothermia induced by chemogenetic inhibition of DMHVgat2 neurons significantly reduced infarct volume and disseminated neuronal injury after stroke. We later investigated the role of DMH-projecting GABAergic ventrolateral preoptic area (vlPOA) neurons in hypothermia-induced neuroprotection against stroke using rAAV-EF1a-double floxed-hChR2(H134R)-EYFP-WPRE-HGHpA (n=10) and rAAV2/1-CAG-DIO-GFP (n=10). The optogenetic modification of DMH-projecting vlPOAVgat2 neurons caused hypothermia and consecutive neuroprotection after stroke. Collectively, we induced therapeutic hypothermia by modulating the GABAergic neurons located at the DMH and DMH-projecting vlPOAVgat2 neurons to induce neuroprotective effects against stroke and promote neuronal survival. Our findings revealed a thermoregulatory projection in the central nervous system and suggested an alternative therapeutic approach against stroke.