The effects of i.p. melatonin (4 + 4 mg/kg, after induction of ischemia and at reperfusion onset) administered either alone or in combination with the thrombolytic tissue‐plasminogen activator (t‐PA, 10 mg/kg), on cerebral laser Doppler flow (LDF) and ischemic injury were studied after 30 min of middle cerebral artery (MCA) thread occlusion in male C57BL/6 mice. Thread occlusions resulted in reproducible focal ischemias, followed by hyperperfusion reactions immediately after thread withdrawal, as revealed by LDF measurements. Compared with animals receiving normal saline (peak LDF after reperfusion: 172.0 ± 24.2%), postischemic LDF was significantly attenuated in animals treated with melatonin (105.1 ± 6.7%, P < 0.05). Delivery of t‐PA (132.8 ± 22.3%) or t‐PA plus melatonin (164.7 ± 36.7%), on the contrary, did not influence postischemic LDF recordings. Twenty‐four hours after reperfusion, melatonin treated mice had significantly increased neuronal survival and decreased disseminate cell injury in the ischemia‐vulnerable striatum, as investigated by cresyl violet and terminal transferase biotinylated‐dUTP nick end labeling stainings. The protective effects were associated with inhibition of caspase‐3 activity. Melatonin administration also increased neuronal survival after 30 min MCA occlusion in animals treated with t‐PA, although t‐PA itself already decreased the degree of injury in a significant manner. Our data demonstrate that melatonin reduces disseminated neuronal injury in the striatum after mild focal ischemia. Brain protection is independent of hemodynamic changes and involves inhibition of caspase‐3.