Responses of microglia in the mouse brain after ischemic stroke

This project investigated responses of microglia in the mouse brain after an ischemic stroke. Mice were subjected to transient focal cerebral ischemia induced by 1 hour of left middle cerebral artery (MCA) occlusion followed by reperfusion. Control mice were subjected to sham surgery without the occlusion of the MCA. Three days after MCA occlusion or sham surgery, microglia were enriched from the brain by fluorescence-activated cell sorting and subjected to bulk RNA-seq. This project was closely related to our previously published dataset GSE141972; therefore, the animals had the same genetic background for comparison purposes. The animals used in this study were hemizygouse for CX3CR1(CreER), so that they can serve as wild-type controls for any conditional knockout driven by the CX3CR1(CreER) promoter. Overall design: Hemizygous CX3CR1(CreER/WT) mice on C57BL/6 background were used. To serve as wild-type controls to other conditional knockout mice driven by the CX3CR1(CreER) promoter, these mice received intraperitoneal injections of tamoxifen (75 mg/kg/day for 4 days). Forteen days after the initial tamoxifen injection,mice (8-10 weeks old, male) were subjected to transient focal cerebral ischemia induced by occlusion of the left middle cerebral artery for 1 hour, followed by reperfusion, or control sham surgery. The CD11b+CD45low (microglia) cell population was collected by fluorescence-activated cell sorting from the left (ipsilesional) brain hemisphere 3 days after ischemic stroke or sham operation. In each group there were 2-3 biological replicates. Each sample was from 1 mouse brain. The sorted cells were subjected to mRNA sequencing. Re-used Samples in GSE141972/PRJNA595477: GSM4217272 SRX7366406 SAMN13556317 WT microglia 1 GSM4217273 SRX7366407 SAMN13556316 WT microglia 2 GSM4217274 SRX7366408 SAMN13556315 WT microglia 3