title: Hippocampus and cerebral cortex present a different autophagic response after oxygen and glucose deprivation in an ex vivo rat brain slice model.
abstract: OBJECTIVE
  To evaluate the neuroprotective role of autophagy in the cerebral cortex and hippocampus using an ex vivo animal model of stroke in brain slices.
  
  
  METHODS
  Brain slices were maintained for 30 min in oxygen and glucose deprivation (OGD) followed by 3 h in normoxic conditions to simulate the reperfusion that follows ischaemia in vivo (RL, reperfusion-like). Phagophore formation (Beclin 1 and LC3B) as well as autophagy flux (p62/SQSTM1, Atg5, Atg7 and polyubiquitin) markers were quantified by Western blot and/or qPCR. The release of lactate dehydrogenase (LDH) and glutamate in the medium was used as a measure of the mortality in the absence and in the presence of the autophagy inhibitor 3-methyladenine.
  
  
  RESULTS
  Striking differences in the autophagy markers were observed between the hippocampus and cerebral cortex in normoxic conditions. OGD/RL induced increases both in the phagophore formation and in the autophagy flux in the first three hours in the cerebral cortex that were not observed in the hippocampus. The blocking of autophagy increased the OGD/RL-induced mortality, increased the glutamate release in both the cerebral cortex and hippocampus and abolished the OGD-induced decrease in the polyubiquitinated proteins in the cerebral cortex.
  
  
  CONCLUSIONS
  We conclude that OGD induces a rapid autophagic response in the cerebral cortex that plays a neuroprotective role. Polyubiquitination levels and control of the glutamate release appear to be involved in the neuroprotective role of autophagy.
authors.affiliation: Área de Biología Celular, Instituto de Biomedicina, Universidad de León, Leon, Spain.
authors.forename: Diego
authors.initials: D
authors.lastname: Pérez-Rodríguez
authors.affiliation: Área de Biología Celular, Instituto de Biomedicina, Universidad de León, Leon, Spain.
authors.forename: Berta
authors.initials: B
authors.lastname: Anuncibay-Soto
authors.affiliation: Área de Biología Celular, Instituto de Biomedicina, Universidad de León, Leon, Spain.
authors.forename: Irene L
authors.initials: IL
authors.lastname: Llorente
authors.affiliation: Área de Medicina y Cirugía Veterinaria, Instituto de Biomedicina, Universidad de León, Leon, Spain.
authors.forename: Carlos C
authors.initials: CC
authors.lastname: Pérez-García
authors.affiliation: Área de Biología Celular, Instituto de Biomedicina, Universidad de León, Leon, Spain.
authors.forename: Arsenio
authors.initials: A
authors.lastname: Fernández-López
mesh_terms: D000818:Animals; D001343:Autophagy; D002545:Brain Ischemia / Q000378:metabolism*; D015687:Cell Hypoxia; D002540:Cerebral Cortex / Q000378:metabolism*; D005947:Glucose / Q000172:deficiency; D018698:Glutamic Acid / Q000378:metabolism; D006624:Hippocampus / Q000378:metabolism*; D066298:In Vitro Techniques; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000378:metabolism; D020521:Stroke / Q000378:metabolism*
chemical_list: D018698:Glutamic Acid; D005947:Glucose
keywords: 3-methyladenine; LC3B; OGD; autophagy; beclin1; mortality