Physical activity prevents stroke and is associated with less severe strokes. The neuroprotective effect in patients treated with intravenous (i.v.) recombinant tissue plasminogen activator (rt-PA), remains uncertain. We aimed at evaluating the relationship between previous physical activity and outcomes in stroke patients treated with i.v. rt-PA. OPHELIE-SPORT was a prospective observational multicenter study conducted in French and Japanese stroke patients treated with i.v. rt-PA. We evaluated the presence, weekly duration (<2, 2–5, >5 h) and intensity (light, moderate, heavy) of previous leisure-time physical activity according to standardized criteria. The primary end-point was an excellent outcome [modified Rankin Scale (mRS) 0–1 or similar to the pre-stroke mRS] after 3 months. Secondary end-points were good outcome (mRS 0–2 or similar to the pre-stroke mRS), and death. Of 519 patients, 74 (14.3 %) had regular physical activity before stroke. They were 14 years younger (p < 0.001), treated 25 min earlier (p = 0.004) and more likely to be men, free of pre-stroke handicap (mRS = 0), atrial fibrillation, arterial hypertension, and diabetes mellitus. National Institutes of Health Stroke Scale scores, at baseline (p = 0.183) and 24 h later (p = 0.203), did not differ between patients with and without physical activity. After adjustment on confounders, there was no association between previous leisure-time physical activity and outcome. Outcomes 3 months after treatment of cerebral ischaemia with i.v. rt-PA are not influenced by previous physical activity.