Focal ischemia in rats causes time-dependent expression of c-fos protein immunoreactivity in widespread regions of ipsilateral cortex

Abstract

c-Fos protein expression was examined in brain by immunohistochemistry following permanent middle cerebral artery (MCA) occlusion above the rhinal fissure and ipsilateral common carotid artery (CCA) occlusion in Long-Evans rats. In sham-operated animals, c-fos protein-like immunoreactivity (CFPLI) was confined to neuronal nuclei of the hypothalamus and was not present in other regions including cerebral cortex. In the core territory of the MCA, CFPLI was not detected when examined at 15 and 30 min, 1, 4 and 8 h and 1, 2, 4 and 7 days after occlusion. Focal ischemia induced two temporal and spatial patterns of CFPLI. At 1 h, c-fos protein was expressed in the nuclei of many neurons in layers II–V of the ipsilateral cortex both immediately adjacent to and remote from the ischemic territory. Within regions outside the MCA territory (e.g. cingulate gyrus and prinform cortices), CFPLI in these neurons peaked at 2–4 and was undetectable after 2 days. Neurons in the zone immediately surrounding the ischemic core within MCA territory also expressed CFPLI, but in contrast, continued to express c-fos up to 4 days after ischemia. Immunoreactivity surrounding the ischemic core was found in neuronal nuclei predominantly, although from 1 to 4 days, CFPLI was found in perikarya and dendrites as well. MK-801 (3 mg/kg, i.p., 30 min prior to occlusion) completely blocked the early c-fos protein induction in all regions but expression within neurons surrounding the ischemic core was present 1 day after a single injection. The initial induction of c-fos protein in cortical neurons distant from the ischemic core lesion and prolonged c-fos expression surrounding the infarct suggests that c-fos expression is linked to both early onset-short lasting stimulation and more persistent excitatory or plasticity-related phenomena associated with focal ischemic brain injury.