Cysteinyl leukotrienes (CysLTs), potent inflammatory mediators, are released from ischemic brain, and may regulate ischemic injury through activating CysLT 1 and CysLT 2 receptors. The CysLT 1 receptor is closely associated with ischemic injury and post-ischemic repair; however, the CysLT 2 receptor-mediated responses remain unknown. Here, we investigated the spatiotemporal profiles and implications of CysLT 2 receptor expression and localization in rat brain after focal cerebral ischemia. CysLT 2 receptors were normally localized in astrocytes in the cortex and around the ventricles. After focal cerebral ischemia, CysLT 2 receptor expression was up-regulated in concert with neuronal and glial responses. In the acute phase (6–24 h), up-regulated CysLT 2 receptors were restricted to injured neurons in the ischemic core; while in the late phase (3–28 days), the up-regulation was restricted to hypertrophic microglia (ischemic core) and mainly localized in hypertrophic astrocytes (boundary zone). Thus, the spatiotemporal profiles of CysLT 2 receptor expression suggest that it plays regulatory roles in acute neuron injury, and astrocytosis and microgliosis in the late phase.