Single-cell RNA sequencing identifies microglial state changes associated with iTBS after ischemia–reperfusion injury

Abstract

Understanding how to effectively treat neurological deficits after stroke remains difficult, as both the therapeutic targets and clinical outcomes are still uncertain. Although intermittent Theta burst stimulation (iTBS) has recently been shown to protect neural tissue in stroke-induced rats, its underlying biology has not been clearly defined. To clarify how iTBS acts at the cellular level, we applied single-cell transcriptome sequencing to a rat model of middle cerebral artery occlusion. After receiving the iTBS protocol, rats underwent behavioral assessments to determine the extent of neurological recovery. We then profiled gene expression across neuronal subtypes using 10x Genomics technology. iTBS markedly alleviated neurological impairment, and single-cell profiling indicated that the stimulation was associated with changes in microglial transcripts (e.g., Vav3 and Cblb) alongside coordinated responses in other cell populations. Together, these data suggest that iTBS is associated with changes in post-ischemic microglial state composition and transcriptional programs, highlighting candidate neuroinflammatory processes for future mechanistic investigation.