title: Dexamethasone enhances necrosis-like neuronal death in ischemic rat hippocampus involving μ-calpain activation.
abstract: Transient forebrain ischemia (TFI) leads to hippocampal CA1 pyramidal cell death which is aggravated by glucocorticoids (GC). It is unknown how GC affect apoptosis and necrosis in cerebral ischemia. We therefore investigated the co-localization of activated caspase-3 (casp-3) with apoptosis- and necrosis-like cell death morphologies in CA1 of rats treated with dexamethasone prior to TFI (DPTI). In addition, apoptosis- (casp-9, casp-3, casp-3-cleaved PARP and cleaved α-spectrin 145/150 and 120kDa) and necrosis-related (calpain-specific casp-9 cleavage, μ-calpain upregulation and cleaved α-spectrin 145/150kDa) cell death mechanisms were investigated by Western blot analysis. DPTI expedited CA1 neuronal death from day 4 to day 1 and increased the magnitude of CA1 neuronal death from 66.2% to 91.3% at day 7. Furthermore, DPTI decreased the overall (days 1-7) percentage of dying neurons displaying apoptosis-like morphology from 4.7% to 0.3% and, conversely, increased the percentage of neurons with necrosis-like morphology from 95.3% to 99.7%. In animals subjected to TFI without dexamethasone (ischemia-only), 7.4% of all dying CA1 neurons were casp-3-immunoreactive (IR), of which 3.1% co-localized with apoptosis-like and 4.3% with necrosis-like changes. By contrast, DPTI decreased the percentage of dying neurons with casp-3 IR to 1.4%, of which 0.3% co-localized with apoptosis-like changes and 1.1% with necrosis-like changes. Western blot analysis from DPTI animals showed a significant elevation of μ-calpain, a calpain-produced necrosis-related casp-9 fragment (25kDa) and cleavage of α-spectrin into 145/150kDa fragments at day 4, whereas in ischemia-only animals a significant increase of casp-3-cleaved PARP, cleavage of α-spectrin into 145/150 and 120kDa fragments was detected at day 7. We conclude that DPTI, in addition to augmenting and expediting CA1 neuronal death, causes a shift from apoptosis-like cell death to necrosis involving μ-calpain activation.
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Ole Maaloesvej 5, Copenhagen 2200, Denmark; Department of Neurology, Donauspital, Langobardenstrasse 122, A-1220 Vienna, Austria. Electronic address: georg.mueller@wienkav.at.
authors.forename: Georg Johannes
authors.initials: GJ
authors.lastname: Müller
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Ole Maaloesvej 5, Copenhagen 2200, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Hasseldam
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Ole Maaloesvej 5, Copenhagen 2200, Denmark.
authors.forename: Rune Skovgaard
authors.initials: RS
authors.lastname: Rasmussen
authors.affiliation: Department of Biomedical Sciences, University of Copenhagen, Ole Maaloesvej 5, Copenhagen 2200, Denmark.
authors.forename: Flemming Fryd
authors.initials: FF
authors.lastname: Johansen
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects; D002154:Calpain / Q000378:metabolism*; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D003907:Dexamethasone / Q000009:adverse effects*; D004195:Disease Models, Animal; D006624:Hippocampus / Q000473:pathology*; D002546:Ischemic Attack, Transient / Q000150:complications / Q000473:pathology*; D008297:Male; D009336:Necrosis / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
chemical_list: D000893:Anti-Inflammatory Agents; D003907:Dexamethasone; D002154:Calpain; D020169:Caspases; C449002:mu-calpain
keywords: Apoptosis-like; Dexamethasone; Glucocorticoids; Hippocampus; Ischemia; Morphology; Necrosis-like; μ-Calpain