title: Tissue inhibitor of metalloproteinases-3 mediates the death of immature oligodendrocytes via TNF-α/TACE in focal cerebral ischemia in mice.
abstract: OBJECTIVE
  Oligodendrocyte (OL) death is important in focal cerebral ischemia. TIMP-3 promotes apoptosis in ischemic neurons by inhibiting proteolysis of TNF-α superfamily of death receptors. Since OLs undergo apoptosis during ischemia, we hypothesized that TIMP-3 contributes to OL death.
  
  
  METHODS
  Middle cerebral artery occlusion (MCAO) was induced in Timp-3 knockout (KO) and wild type (WT) mice with 24 or 72 h of reperfusion. Cell death in white matter was investigated by stereology and TUNEL. Mature or immature OLs were identified using antibodies against glutathione S-transferase-π (GST-π) and galactocerebroside (GalC), respectively. Expression and level of proteins were examined using immunohistochemistry and immunoblotting. Protein activities were determined using a FRET peptide.
  
  
  RESULTS
  Loss of OL-like cells was detected at 72 h only in WT ischemic white matter where TUNEL showed greater cell death. TIMP-3 expression was increased in WT reactive astrocytes. GST-π was reduced in ischemic white matter of WT mice compared with WT shams with no difference between KO and WT at 72 h. GalC level was significantly increased in both KO and WT ischemic white matter at 72 h. However, the increase in GalC in KO mice was significantly higher than WT; most TUNEL-positive cells in ischemic white matter expressed GalC, suggesting TIMP-3 deficiency protects the immature OLs from apoptosis. There were significantly higher levels of cleaved caspase-3 at 72 h in WT white matter than in KO. Greater expression of MMP-3 and -9 was seen in reactive astrocytes and/or microglia/macrophages in WT at 72 h. We found more microglia/macrophages in WT than in KO, which were the predominant source of increased TNF-α detected in the ischemic white matter. TACE activity was significantly increased in ischemic WT white matter, which was expressed in active microglia/macrophages and OLs.
  
  
  CONCLUSIONS
  Our results suggested that focal ischemia leads to proliferation of immature OLs in white matter and that TIMP-3 contributes to a caspase-3-dependent immature OL death via TNF-α-mediated neuroinflammation. Future studies will be needed to delineate the role of MMP-3 and MMP-9 that were increased in the Timp-3 wild type.
authors.affiliation: Department of Neurology, University of New Mexico Health Sciences Center, Albuquerque, NM 87131, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Yang
authors.forename: Fakhreya Y
authors.initials: FY
authors.lastname: Jalal
authors.forename: Jeffrey F
authors.initials: JF
authors.lastname: Thompson
authors.forename: Espen J
authors.initials: EJ
authors.lastname: Walker
authors.forename: Eduardo
authors.initials: E
authors.lastname: Candelario-Jalil
authors.forename: Lu
authors.initials: L
authors.lastname: Li
authors.forename: Ross R
authors.initials: RR
authors.lastname: Reichard
authors.forename: Chi
authors.initials: C
authors.lastname: Ben
authors.forename: Qing-Xiang
authors.initials: QX
authors.lastname: Sang
authors.forename: Lee Anna
authors.initials: LA
authors.lastname: Cunningham
authors.forename: Gary A
authors.initials: GA
authors.lastname: Rosenberg
mesh_terms: D051722:ADAM Proteins / Q000378:metabolism*; D000072198:ADAM17 Protein; D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D002545:Brain Ischemia / Q000473:pathology / Q000503:physiopathology*; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000502:physiology*; D031541:Fluorescence Resonance Energy Transfer; D020287:In Situ Nick-End Labeling; D020244:Infarction, Middle Cerebral Artery / Q000378:metabolism; D008297:Male; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D009836:Oligodendroglia / Q000166:cytology / Q000473:pathology / Q000502:physiology*; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000235:genetics / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*
chemical_list: D019717:Tissue Inhibitor of Metalloproteinase-3; D014409:Tumor Necrosis Factor-alpha; D053148:Caspase 3; D051722:ADAM Proteins; D019278:Matrix Metalloproteinase 3; D020780:Matrix Metalloproteinase 9; D000072198:ADAM17 Protein; C000606463:Adam17 protein, mouse