Although microhemorrhages are common in the brain of the elderly, the direct impact of these lesions on neural function remains unclear. In this work, we used femtosecond laser irradiation to rupture the wall of single arterioles in the brain of anesthetized rodents, producing a hematoma of similar to 100-mu m diameter. Our objective was to study the impact of these microhemorrhages on cortical activity using cell-resolved two-photon imaging of bulk-loaded calcium-sensitive dye. We monitored peripheral sensory stimulus-induced calcium transients from individual neuronal cell bodies, regions of neuropil, and astrocytes at different distances from the microhemorrhage before and 0.5, 2, and 4 hours after the creation of the lesion. We found that immediately after the hemorrhage the average amplitude of the stimulus-induced calcium response was reduced to about half within 150 mu m from the hematoma. Beyond 300 mu m, there was little effect on cell response, with a smooth increase in response amplitude from 150 mu m to 300 mu m from the lesion. Cortical function gradually improved with time and by four hours after the lesion the response from neurons and astrocytes had recovered to baseline everywhere but within 150 mu m from the hematoma. To assess whether the cells closest to the microhemorrhage recovered over a longer timeframe, we developed a re-openable chronic cranial window preparation that allowed reinjection of calcium-sensitive fluorescent dye. We found that the response largely recovered by one day after the microhemorrhage even within 150 mu m from the hematoma. This work suggests that neuronal and astrocyte function is transiently lost near a microhemorrhage, but recovers within one day after the lesion.
