Microglial depletion abolishes ischemic preconditioning in white matter

Abstract Ischemic preconditioning (IPC) is a phenomenon whereby a brief, non‐injurious ischemic exposure enhances tolerance to a subsequent ischemic challenge. The mechanism of IPC has mainly been studied in rodent stroke models where gray matter (GM) constitutes about 85% of the cerebrum. In humans, white matter (WM) is 50% of cerebral volume and is a critical component of stroke damage. We developed a novel CNS WM IPC model using the mouse optic nerve (MON) and identified the involved immune signaling pathways. Here we tested the hypothesis that microglia are necessary for WM IPC. Microglia were depleted by treatment with the colony stimulating factor 1 receptor (CSF1R) inhibitor PLX5622. MONs were exposed to transient ischemia in vivo, acutely isolated 72 h later, and subjected to oxygen–glucose deprivation (OGD) to simulate a severe ischemic injury (i.e., stroke). Functional and structural axonal recovery was assessed by recording compound action potentials (CAPs) and by microscopy using quantitative stereology. Microglia depletion eliminated IPC‐mediated protection. In control mice, CAP recovery was improved in preconditioned MONs compared with non‐preconditioned MONs, however, in PLX5622‐treated mice, we observed no difference in CAP recovery between preconditioned and non‐preconditioned MONs. Microgliadepletion also abolished IPC protective effects on axonal integrity and survival of mature (APC+) oligodendrocytes after OGD. IPC‐mediated protection was independent of retinal injury suggesting it results from mechanistic processes intrinsic to ischemia‐exposed WM. We conclude that preconditioned microglia are critical for IPC in WM. The “preconditioned microglia” phenotype might protect against other CNS pathologies and is a neurotherapeutic horizon worth exploring.