Apoptotic and autophagic cell death have been implicated, on the basis of morphological and biochemical criteria, in neuronal loss occurring in neurodegenerative diseases and it has been shown that they may overlap. We have studied the relationship between apoptosis and autophagic cell death in cerebellar granule cells (CGCs) undergoing apoptosis following serum and potassium deprivation. We found that apoptosis is accompanied by an early and marked proliferation of autophagosomal-lysosomal compartments as detected by electron microscopy and immunofluorescence analysis. Autophagy is blocked by hrIGF-1 and forskolin, two well-known inhibitors of CGC apoptosis, as well as by adenovirus-mediated overexpression of Bcl-2. 3-Methyladenine (3-MA) an inhibitor of autophagy, not only arrests this event but it also blocks apoptosis. The neuroprotective effect of 3-MA is accompanied by block of cytochrome c (cyt c) release in the cytosol and by inhibition of caspase-3 activation which, in turn, appears to be mediated by cathepsin B, as CA074-Me, a selective inhibitor of this enzyme, fully blocks the processing of pro-caspase-3. Immunofluorescence analysis demonstratesd that cathepsin B, normally confined inside the lysosomal-endosomal compartment, is released during apoptosis into the cytosol where this enzyme may act as an execution protease. Collectively, these observations indicate that autophagy precedes and is causally connected with the subsequent onset of programmed death.

Canu, N., Tufi, R., Serafino, A., Amadoro, G., Ciotti, M., Calissano, P. (2005). Role of the autophagic-lysosomal system on low potassium-induced apoptosis in cultured cerebellar granule cells. JOURNAL OF NEUROCHEMISTRY, 92(5), 1228-1242 [10.1111/j.1471-4159.2004.02956.x].

Role of the autophagic-lysosomal system on low potassium-induced apoptosis in cultured cerebellar granule cells

CANU, NADIA
;
CALISSANO, PIETRO
2005-01-01

Abstract

Apoptotic and autophagic cell death have been implicated, on the basis of morphological and biochemical criteria, in neuronal loss occurring in neurodegenerative diseases and it has been shown that they may overlap. We have studied the relationship between apoptosis and autophagic cell death in cerebellar granule cells (CGCs) undergoing apoptosis following serum and potassium deprivation. We found that apoptosis is accompanied by an early and marked proliferation of autophagosomal-lysosomal compartments as detected by electron microscopy and immunofluorescence analysis. Autophagy is blocked by hrIGF-1 and forskolin, two well-known inhibitors of CGC apoptosis, as well as by adenovirus-mediated overexpression of Bcl-2. 3-Methyladenine (3-MA) an inhibitor of autophagy, not only arrests this event but it also blocks apoptosis. The neuroprotective effect of 3-MA is accompanied by block of cytochrome c (cyt c) release in the cytosol and by inhibition of caspase-3 activation which, in turn, appears to be mediated by cathepsin B, as CA074-Me, a selective inhibitor of this enzyme, fully blocks the processing of pro-caspase-3. Immunofluorescence analysis demonstratesd that cathepsin B, normally confined inside the lysosomal-endosomal compartment, is released during apoptosis into the cytosol where this enzyme may act as an execution protease. Collectively, these observations indicate that autophagy precedes and is causally connected with the subsequent onset of programmed death.
2005
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/09 - FISIOLOGIA
English
Con Impact Factor ISI
Apoptosis; Autophagy; Caspase activation; Cathepsin B; Cerebellar granule neurons; Neurodegeneration
3 methyladenine; adenovirus vector; caspase 3; cathepsin B; cytochrome c; enzyme inhibitor; epoxysuccinylisoleucylproline propylamide methyl ester; forskolin; potassium; procaspase 3; protein bcl 2; recombinant somatomedin C; unclassified drug; animal cell; apoptosis; article; autophagy; cell death; cell loss; cerebellum; controlled study; degenerative disease; electron microscopy; enzyme activation; enzyme inhibition; enzyme release; gene overexpression; granule cell; immunofluorescence; lysosome; nerve cell culture; neuroprotection; nonhuman; potassium cell level; priority journal; rat; Adenine; Amino Acid Chloromethyl Ketones; Animals; Animals, Newborn; Antigens, CD; Apoptosis; Autophagy; Caspases; Cathepsins; Cell Size; Cell Survival; Cells, Cultured; Cerebellum; Coumarins; Cysteine Proteinase Inhibitors; DNA-Binding Proteins; Dose-Response Relationship, Drug; Drug Interactions; Erythroid-Specific DNA-Binding Factors; Extracellular Signal-Regulated MAP Kinases; Fluorescent Antibody Technique; Forskolin; Gene Expression Regulation; Glycoside Hydrolases; Green Fluorescent Proteins; Lysosome-Associated Membrane Glycoproteins; Lysosomes; Microscopy, Electron; Microtubule Proteins; Neurons; Oligopeptides; Potassium; Proto-Oncogene Proteins c-bcl-2; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Teprotide; Time Factors; Transcription Factors; Adenoviridae
Canu, N., Tufi, R., Serafino, A., Amadoro, G., Ciotti, M., Calissano, P. (2005). Role of the autophagic-lysosomal system on low potassium-induced apoptosis in cultured cerebellar granule cells. JOURNAL OF NEUROCHEMISTRY, 92(5), 1228-1242 [10.1111/j.1471-4159.2004.02956.x].
Canu, N; Tufi, R; Serafino, A; Amadoro, G; Ciotti, M; Calissano, P
Articolo su rivista
File in questo prodotto:
File Dimensione Formato  
Canu Nadia JNC.pdf

accesso aperto

Descrizione: PDF articolo
Licenza: Non specificato
Dimensione 1.02 MB
Formato Adobe PDF
1.02 MB Adobe PDF Visualizza/Apri

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/56772
Citazioni
  • ???jsp.display-item.citation.pmc??? 36
  • Scopus 122
  • ???jsp.display-item.citation.isi??? 115
social impact