ABSTRACT Microglia cells are inflammatory cells of the central nervous system. Following a stroke, cerebral ischemia activates the microglia cells into a diverse range of phenotypes, each with different physical characteristics, surface markers and secretory products, to mediate, augment or suppress the inflammatory process in the brain. We hope to characterize the behavior of these cells to elucidate their role in the inflammatory process. We developed a middle cerebral artery obstruction (MCAO) model on male Sprague-Dawley rats (250-280g) to simulate transient cerebral ischemia by inserting a 4-0 silicone-coated suture through the external carotid artery and the internal carotid artery to occlude the middle carotid artery. After 60 minutes, the filament was removed to allow for reperfusion. Functional assessments were performed pre-surgery, and on days 1, 2, 3, 5, and 7 post-surgery. The animals were subsequently sacrificed. Their brains were double-stained with immunofluorescent Tmem119 and Iba1 antibodies for immunofluorescence analysis. The results were compared to a sham group which received the same surgery bar the insertion of suture. The MCAO model produced consistent neurological deficits in the rats. Immunofluorescence staining showed that the number of microglia cells, the length of microglia cell processes and the surface area occupied by microglia cells fluctuated post- MCAO surgery but did not show any trends. The changes in the physical characteristics of the microglia cells suggest that there is a change of microglia cell phenotypes with time following transient focal cerebral ischemia. We described how future research can better characterize these changes and how RNA profiling and Western Blotting can be used to identify and define the microglia cell phenotypes present following transient focal cerebral ischemia.
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