title: Ischemic Preconditioning in White Matter: Magnitude and Mechanism.
abstract: Ischemic preconditioning (IPC) is a robust neuroprotective phenomenon whereby brief ischemic exposure confers tolerance to a subsequent ischemic challenge. IPC has not been studied selectively in CNS white matter (WM), although stroke frequently involves WM. We determined whether IPC is present in WM and, if so, its mechanism. We delivered a brief in vivo preconditioning ischemic insult (unilateral common carotid artery ligation) to 12- to 14-week-old mice and determined WM ischemic vulnerability [oxygen-glucose deprivation (OGD)] 72 h later, using acutely isolated optic nerves (CNS WM tracts) from the preconditioned (ipsilateral) and control (contralateral) hemispheres. Functional and structural recovery was assessed by quantitative measurement of compound action potentials (CAPs) and immunofluorescent microscopy. Preconditioned mouse optic nerves (MONs) showed better functional recovery after OGD than the non-preconditioned MONs (31 ± 3 vs 17 ± 3% normalized CAP area, p < 0.01). Preconditioned MONs also showed improved axon integrity and reduced oligodendrocyte injury compared with non-preconditioned MONs. Toll-like receptor-4 (TLR4) and type 1 interferon receptor (IFNAR1), key receptors in innate immune response, are implicated in gray matter preconditioning. Strikingly, IPC-mediated WM protection was abolished in both TLR4(-/-) and IFNAR1(-/-) mice. In addition, IPC-mediated protection in WM was also abolished in IFNAR1(fl/fl) LysM(cre), but not in IFNAR1(fl/fl) control, mice. These findings demonstrated for the first time that IPC was robust in WM, the phenomenon being intrinsic to WM itself. Furthermore, WM IPC was dependent on innate immune cell signaling pathways. Finally, these data demonstrated that microglial-specific expression of IFNAR1 plays an indispensable role in WM IPC.
  
  
  CONCLUSIONS
  Ischemic preconditioning (IPC) has been studied predominantly in gray matter, but stroke in humans frequently involves white matter (WM) as well. Here we describe a novel, combined in vivo/ex vivo mouse model to determine whether IPC occurs in WM. It does. Using genetically altered mice, we identified two innate immune cell receptors, Toll-like receptor 4 and type 1 interferon receptor (IFNAR1), that are required for IPC-mediated protection in WM. Furthermore, using microglia-targeted IFNAR1 knockdown, we demonstrate that interferon signaling specifically in microglia is essential for this protection. The discovery of IPC as an intrinsic capability of WM is novel and important. This is also the first in vivo demonstration that cell-type-specific expression of an individual gene plays an indispensable role in IPC-mediated protection.
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Margaret A
authors.identifier: http://orcid.org/0000-0002-2045-5454
authors.initials: MA
authors.lastname: Hamner
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Zucheng
authors.identifier: http://orcid.org/0000-0001-5851-3261
authors.initials: Z
authors.lastname: Ye
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Richard V
authors.identifier: http://orcid.org/0000-0003-3386-8212
authors.initials: RV
authors.lastname: Lee
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Jamie R
authors.initials: JR
authors.lastname: Colman
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Thu
authors.initials: T
authors.lastname: Le
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Davin C
authors.initials: DC
authors.lastname: Gong
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195 bransom@uw.edu.
authors.forename: Bruce R
authors.identifier: http://orcid.org/0000-0002-9658-0294
authors.initials: BR
authors.lastname: Ransom
authors.affiliation: Department of Neurology, University of Washington, Seattle, Washington 98195.
authors.forename: Jonathan R
authors.identifier: http://orcid.org/0000-0001-6080-9758
authors.initials: JR
authors.lastname: Weinstein
mesh_terms: D000818:Animals; D002545:Brain Ischemia / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D019194:Ischemic Preconditioning / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D053633:Receptor, Interferon alpha-beta / Q000096:biosynthesis* / Q000235:genetics; D051197:Toll-Like Receptor 4 / Q000096:biosynthesis* / Q000235:genetics; D066127:White Matter / Q000378:metabolism* / Q000473:pathology
chemical_list: C508032:Ifnar1 protein, mouse; C493487:Tlr4 protein, mouse; D051197:Toll-Like Receptor 4; D053633:Receptor, Interferon alpha-beta
keywords: interferon; ischemic preconditioning; microglia; toll-like receptor-4; white matter