Document Detail


Role of the autophagic-lysosomal system on low potassium-induced apoptosis in cultured cerebellar granule cells.
MedLine Citation:
PMID:  15715672     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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 demonstrated 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.
Authors:
Nadia Canu; Roberta Tufi; Anna Lucia Serafino; Giuseppina Amadoro; Maria Teresa Ciotti; Pietro Calissano
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  92     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  2005 Mar 
Date Detail:
Created Date:  2005-02-17     Completed Date:  2005-04-28     Revised Date:  2009-11-19    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  England    
Other Details:
Languages:  eng     Pagination:  1228-42     Citation Subset:  IM    
Affiliation:
Dipartimento di Neuroscienze, Facoltà di Medicina e Chirurgia, Università di Tor Vergata, Roma, Italia. n.canu@inmm.cnr.it
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MeSH Terms
Descriptor/Qualifier:
Adenine / analogs & derivatives*,  pharmacology
Amino Acid Chloromethyl Ketones / pharmacology
Animals
Animals, Newborn
Antigens, CD / metabolism
Apoptosis / drug effects*
Autophagy / physiology*
Caspases / metabolism
Cathepsins / metabolism
Cell Size / drug effects
Cell Survival / drug effects
Cells, Cultured
Cerebellum / cytology*
Coumarins / pharmacology
Cysteine Proteinase Inhibitors / pharmacology
DNA-Binding Proteins / pharmacology
Dose-Response Relationship, Drug
Drug Interactions
Erythroid-Specific DNA-Binding Factors
Extracellular Signal-Regulated MAP Kinases / metabolism
Fluorescent Antibody Technique / methods
Forskolin / pharmacology
Gene Expression Regulation
Glycoside Hydrolases / metabolism
Green Fluorescent Proteins / metabolism
Lysosome-Associated Membrane Glycoproteins
Lysosomes / physiology*,  ultrastructure
Microscopy, Electron / methods
Microtubule Proteins / metabolism
Neurons / drug effects*,  ultrastructure
Oligopeptides / pharmacology
Potassium / pharmacology*
Proto-Oncogene Proteins c-bcl-2 / metabolism
RNA, Messenger / biosynthesis
Rats
Rats, Wistar
Reverse Transcriptase Polymerase Chain Reaction / methods
Teprotide / pharmacology
Time Factors
Transcription Factors / pharmacology
Chemical
Reg. No./Substance:
0/Ac-aspartyl-glutamyl-valyl-aspartyl-aminomethylcoumarin; 0/Amino Acid Chloromethyl Ketones; 0/Antigens, CD; 0/Coumarins; 0/Cysteine Proteinase Inhibitors; 0/DNA-Binding Proteins; 0/Erythroid-Specific DNA-Binding Factors; 0/Lysosome-Associated Membrane Glycoproteins; 0/Microtubule Proteins; 0/Oligopeptides; 0/Proto-Oncogene Proteins c-bcl-2; 0/RNA, Messenger; 0/Transcription Factors; 0/acetyl-tyrosyl-valyl-alanyl-aspartyl-7-amino-4-methylcoumarinamide; 0/benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; 147336-22-9/Green Fluorescent Proteins; 35115-60-7/Teprotide; 5142-23-4/3-methyladenine; 66428-89-5/Forskolin; 73-24-5/Adenine; 7440-09-7/Potassium; EC 2.7.11.24/Extracellular Signal-Regulated MAP Kinases; EC 3.2.1.-/Glycoside Hydrolases; EC 3.2.1.-/beta-galactanase; EC 3.4.-/Cathepsins; EC 3.4.22.-/Caspases

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