title: Microglial activation precedes and predominates over macrophage infiltration in transient focal cerebral ischemia: a study in green fluorescent protein transgenic bone marrow chimeric mice.
abstract: Resident microglia and hematogenous macrophages play crucial roles in the pathogenetic cascade following cerebral ischemia but may functionally differ regarding neuroprotective and cytotoxic properties. Distinction between these cells has not been possible due to a lack of discriminating cellular markers. We generated bone marrow chimeric mice by transplanting bone marrow from green fluorescent protein (GFP) transgenic mice into irradiated wild-type recipients. Transient focal cerebral ischemia was induced by transient middle cerebral artery occlusion (MCAO) for 30 min. Resident microglia and infiltrating macrophages were identified by immunohistochemistry and GFP fluorescence after 1-28 days. The first blood-derived cells infiltrating the infarct area were seen on Day 1 and identified as granulocytes. Hematogenous GFP(+) macrophages were rarely observed on Day 2, reached peak numbers on Day 7, and decreased thereafter. In contrast, resident GFP(-) microglial cells rapidly became activated already on Day 1 after MCAO. Even on Days 4 and 7, most macrophage-like cells remained GFP(-), indicating their derivation from resident microglia. Hematogenous macrophages were able to acquire a ramified morphology indistinguishable from resident microglia while microglial cells could develop into a phagocytic phenotype indistinguishable from infiltrating macrophages. The vast majority of macrophages in the infarct area are derived from local microglia, revealing a remarkable predominance of local defense mechanisms over immune cells arriving from the blood. GFP bone marrow chimeric mice are a powerful tool to further differentiate the function of resident microglia and hematogenous macrophages following cerebral ischemia.
authors.affiliation: Department of Neurology, Universitätsklinikum Münster, Münster, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Schilling
authors.forename: Michael
authors.initials: M
authors.lastname: Besselmann
authors.forename: Christine
authors.initials: C
authors.lastname: Leonhard
authors.forename: Marcus
authors.initials: M
authors.lastname: Mueller
authors.forename: E Bernd
authors.initials: EB
authors.lastname: Ringelstein
authors.forename: Reinhard
authors.initials: R
authors.lastname: Kiefer
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000473:pathology; D016026:Bone Marrow Transplantation; D001921:Brain / Q000098:blood supply / Q000473:pathology / Q000503:physiopathology; D002465:Cell Movement / Q000502:physiology; D004195:Disease Models, Animal; D018450:Disease Progression; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D002546:Ischemic Attack, Transient / Q000473:pathology* / Q000503:physiopathology; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008264:Macrophages / Q000473:pathology* / Q000502:physiology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D017628:Microglia / Q000378:metabolism / Q000473:pathology*; D011828:Radiation Chimera
chemical_list: D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins