title: Calpain-mediated cleavage of Beclin-1 and autophagy deregulation following retinal ischemic injury in vivo.
abstract: Autophagy is the major intracellular degradation pathway that regulates long-lived proteins and organelles turnover. This process occurs at basal levels in all cells but it is rapidly upregulated in response to starvation and cellular stress. Although being recently implicated in neurodegeneration, it remains still unclear whether autophagy has a detrimental or protective role. In this study, we investigated the dynamics of the autophagic process in retinal tissue that has undergone transient ischemia, an experimental model that recapitulates features of ocular pathologies, including glaucoma, anterior ischemic optic neuropathy and retinal vessels occlusion. Retinal ischemia, induced in adult rats by increasing the intraocular pressure, was characterized by a reduction in the phosphatidylethanolamine-modified form of LC3 (LC3II) and by a significant decrease in Beclin-1. The latter event was associated with a proteolytic cleavage of Beclin-1, leading to the accumulation of a 50-kDa fragment. This event was prevented by intravitreal treatment with the non-competitive N-methyl-D-aspartate antagonist MK801 and calpain inhibitors or by calpain knockdown. Blockade of autophagy by pharmacological inhibition or Beclin-1 silencing in RGC-5 increased cell death, suggesting a pro-survival role of the autophagic process in this neuronal cell type. Altogether, our results provide original evidence for calpain-mediated cleavage of Beclin-1 and deregulation of basal autophagy in the rat retina that has undergone ocular ischemia/reperfusion injury.
authors.affiliation: Department of Pharmacobiology, University of Calabria, Arcavacata di Rende, Italy. rossella.russo@unical.it
authors.forename: R
authors.initials: R
authors.lastname: Russo
authors.forename: L
authors.initials: L
authors.lastname: Berliocchi
authors.forename: A
authors.initials: A
authors.lastname: Adornetto
authors.forename: G P
authors.initials: GP
authors.lastname: Varano
authors.forename: F
authors.initials: F
authors.lastname: Cavaliere
authors.forename: C
authors.initials: C
authors.lastname: Nucci
authors.forename: D
authors.initials: D
authors.lastname: Rotiroti
authors.forename: L A
authors.initials: LA
authors.lastname: Morrone
authors.forename: G
authors.initials: G
authors.lastname: Bagetta
authors.forename: M T
authors.initials: MT
authors.lastname: Corasaniti
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism*; D001343:Autophagy; D000071186:Beclin-1; D002154:Calpain / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004195:Disease Models, Animal; D006801:Humans; D007511:Ischemia / Q000235:genetics / Q000378:metabolism; D007668:Kidney / Q000098:blood supply* / Q000378:metabolism; D008297:Male; D011499:Protein Processing, Post-Translational; D051381:Rats; D017208:Rats, Wistar; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism; D015427:Reperfusion Injury / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology
chemical_list: D051017:Apoptosis Regulatory Proteins; D000071186:Beclin-1; C512829:Becn1 protein, rat; D016194:Receptors, N-Methyl-D-Aspartate; D002154:Calpain