000199618 001__ 199618
000199618 005__ 20180913062531.0
000199618 0247_ $$2doi$$a10.4161/auto.28264
000199618 022__ $$a1554-8627
000199618 02470 $$2ISI$$a000335433700010
000199618 037__ $$aARTICLE
000199618 245__ $$aInvolvement of autophagy in hypoxic-excitotoxic neuronal death
000199618 260__ $$aAustin$$bLandes Bioscience$$c2014
000199618 269__ $$a2014
000199618 300__ $$a15
000199618 336__ $$aJournal Articles
000199618 520__ $$aNeuronal autophagy is increased in numerous excitotoxic conditions including neonatal cerebral hypoxia-ischemia (HI). However, the role of this HI-induced autophagy remains unclear. To clarify this role we established an in vitro model of excitotoxicity combining kainate treatment (Ka, 30 mu M) with hypoxia (Hx, 6% oxygen) in primary neuron cultures. KaHx rapidly induced excitotoxic death that was completely prevented by MK801 or EGTA. KaHx also stimulated neuronal autophagic flux as shown by a rise in autophagosome number (increased levels of LC3-II and punctate LC3 labeling) accompanied by increases in lysosomal abundance and activity (increased SQSTM1/p62 degradation, and increased LC3-II levels in the presence of lysosomal inhibitors) and fusion (shown using an RFP-GFP-LC3 reporter). To determine the role of the enhanced autophagy we applied either pharmacological autophagy inhibitors (3-methyladenine or pepstatinA/E64) or lentiviral vectors delivering shRNAs targeting Becn1 or Atg7. Both strategies reduced KaHx-induced neuronal death. A prodeath role of autophagy was also confirmed by the enhanced toxicity of KaHx in cultures overexpressing BECN1 or ATG7. Finally, in vivo inhibition of autophagy by intrastriatal injection of a lentiviral vector expressing a Becn1-targeting shRNA increased the volume of intact striatum in a rat model of severe neonatal cerebral HI. These results clearly show a death-mediating role of autophagy in hypoxic-excitotoxic conditions and suggest that inhibition of autophagy should be considered as a neuroprotective strategy in HI brain injuries.
000199618 6531_ $$aautophagy
000199618 6531_ $$aneonatal hypoxia-ischemia
000199618 6531_ $$aneuroprotection
000199618 6531_ $$aneuronal death
000199618 6531_ $$aexcitotoxicity
000199618 700__ $$aGinet, Vanessa$$uUniv Lausanne, Fac Biol & Med, Dept Fundamental Neurosci, Lausanne, Switzerland
000199618 700__ $$aSpiehlmann, Amelie$$uUniv Lausanne, Fac Biol & Med, Dept Fundamental Neurosci, Lausanne, Switzerland
000199618 700__ $$aRummel, Coralie$$uUniv Lausanne, Fac Biol & Med, Dept Fundamental Neurosci, Lausanne, Switzerland
000199618 700__ $$aRudinskiy, Nikita$$uEcole Polytech Fed Lausanne, Brain Mind Inst, Lausanne, Switzerland
000199618 700__ $$aGrishchuk, Yulia$$uUniv Lausanne, Fac Biol & Med, Dept Fundamental Neurosci, Lausanne, Switzerland
000199618 700__ $$0240509$$aLuthi-Carter, Ruth$$g158211$$uEcole Polytech Fed Lausanne, Brain Mind Inst, Lausanne, Switzerland
000199618 700__ $$aClarke, Peter G. H.$$uUniv Lausanne, Fac Biol & Med, Dept Fundamental Neurosci, Lausanne, Switzerland
000199618 700__ $$aTruttmann, Anita C.$$uUniv Lausanne Hosp, Clin Neonatol, Dept Pediat & Pediat Surg, Lausanne, Switzerland
000199618 700__ $$aPuyal, Julien$$uUniv Lausanne, Fac Biol & Med, Dept Fundamental Neurosci, Lausanne, Switzerland
000199618 773__ $$j10$$k5$$q846-860$$tAutophagy
000199618 909C0 $$0252338$$pLNGF$$xU10838
000199618 909CO $$ooai:infoscience.tind.io:199618$$particle
000199618 937__ $$aEPFL-ARTICLE-199618
000199618 973__ $$aEPFL$$rREVIEWED$$sPUBLISHED
000199618 980__ $$aARTICLE