Role of calpain- and interleukin-1 beta converting enzyme-like proteases in the beta-amyloid-induced death of rat hippocampal neurons in culture.
We investigated the potential role of different proteases in the death of cultured rat hippocampal pyramidal neurons induced by beta-amyloid (A beta) (25-35). Both A beta(25-35)- and staurosporine-induced death of these neurons appeared to involve apoptosis, as indicated using Hoechst 33342 and terminal dUDP nick end labeling staining, whereas NMDA-induced death appeared more complex. Two irreversible inhibitors of the interleukin-1 beta converting enzyme (ICE) and related proteases, Z-Val-Ala-Asp-CH2F and acetyl-Tyr-Val-Ala-Asp-chloromethyl ketone, blocked neuronal death produced by A beta(25-35), staurosporine, and NMDA to differing extents. Furthermore, MDL 28,170, a selective inhibitor of the calcium-regulated protease calpain, also inhibited death induced by all agents. A beta(25-35) and staurosporine stimulated the breakdown of the protein spectrin, a calpain substrate. Spectrin breakdown was inhibited by MDL 28,170 but not by ICE inhibitors. Leupeptin was only effective in preventing NMDA-induced death. These results support the role of apoptosis in neuronal death due to A beta(25-35) treatment and also suggest a role for calcium-regulated proteases in this process.
Jordán
J
J
Department of Pharmacological and Physiological Sciences, University of Chicago, Illinois 60637, USA.
Galindo
M F
MF
Miller
R J
RJ
D016229:Amyloid beta-Peptides / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000502:physiology; D002154:Calpain / Q000378:metabolism*; D020170:Caspase 1; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects / Q000201:enzymology; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D053938:DNA Fragmentation; D010450:Endopeptidases / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018690:Excitatory Amino Acid Agonists / Q000494:pharmacology; D005260:Female; D007375:Interleukin-1 / Q000378:metabolism; D016202:N-Methylaspartate / Q000494:pharmacology; D009498:Neurotoxins / Q000494:pharmacology; D010446:Peptide Fragments / Q000494:pharmacology; D011247:Pregnancy; D017966:Pyramidal Cells / Q000166:cytology* / Q000187:drug effects* / Q000201:enzymology; D051381:Rats; D019311:Staurosporine / Q000494:pharmacology
D016229:Amyloid beta-Peptides; D004791:Enzyme Inhibitors; D018690:Excitatory Amino Acid Agonists; D007375:Interleukin-1; D009498:Neurotoxins; D010446:Peptide Fragments; D016202:N-Methylaspartate; D010450:Endopeptidases; D002154:Calpain; D003546:Cysteine Endopeptidases; D020170:Caspase 1; D019311:Staurosporine; D002118:Calcium