The study focused on GATAD1 (GATA zinc finger domain-containing 1), a transcription factor important for cardiovascular function, and its negative role in ischemic stroke by worsening blood-brain barrier (BBB) dysfunction through caveolae-mediated transcytosis in endothelial cells. In a mouse model, increased GATAD1 expression was observed in cerebral endothelial cells, while mice lacking GATAD1 in these cells had reduced infarct volumes, less BBB leakage, and better neurological outcomes. With the help of a miniature two-photon microscope (FHIRMTPM V2.0), BBB permeability was assessed in freely-moving mice. The research indicates that GATAD1 promotes BBB permeability by enhancing CD36 expression, a pathway involved in endothelial transcytosis during ischemia. Targeting GATAD1 may offer a new therapeutic approach for ischemic stroke.