Inhibition of microglial SLC2A5 attenuates brain injury following acute ischemic stroke [scRNA-Seq]

The Slc2a5 in microglia plays a crucial role in the brain's injury response. In this study, we utilized single-cell RNA sequencing to examine the impact of Slc2a5 knockout on cell type-specific transcription in the brain. Additionally, we employed single-nucleus RNA sequencing to investigate the cell type-specific transcriptional changes in the brain 24 hours after transient middle cerebral artery occlusion (tMCAO) in response to ischemic stroke. Furthermore, RNA sequencing was conducted to analyze the effects of oxygen-glucose deprivation (OGD) conditions on the transcriptional profiles of wild-type and Slc2a5 knockout microglia in vitro. Overall design: (1)brain cortex were isolated for scRNA-seq from P35-40 widetype and Slc2a5 knockout mice. (2) Brain samples were harvested from widetype and Slc2a5 knockout mice with 24 hours after tMCAO, followed by library preparation using standard 10x Genomics workflows and snRNAseq (NovaSeq 6000, Illumina). Left and right hemispheres were sequenced separately. Hence, we obtained datasets from the contralateral and ipsilateral to ischemic lesion.(3) brain microglia was isolated by FACS from P35-40 widetype and Slc2a5 knockout mice, and culture them under oxygen-glucose deprivation(OGD) condition with or with 24 hours recovery.The library for sequencing was prepared following the protocol for SMARTer Stranded Total RNA-Seq Kit v2.