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 Previous Article | Next Article 
Volume 16, Number 15,
Issue of August 1, 1996
pp. 4696-4706
Copyright ©1996 Society for Neuroscience
Potassium Deprivation-Induced Apoptosis of Cerebellar Granule
Neurons: A Sequential Requirement for New mRNA and Protein
Synthesis, ICE-Like Protease Activity, and Reactive Oxygen
Species
Received Feb. 8, 1996; revised April 18, 1996; accepted May 13, 1996.
Jörg B. Schulz,
Michael Weller, and
Thomas Klockgether
Department of Neurology, University of Tübingen, D-72076
Tübingen, Germany
Potassium (K+) deprivation-induced apoptosis
of cerebellar granule neurons requires new mRNA and protein synthesis.
Using a fluorogenic substrate for interleukin-1 converting enzyme
(ICE), we show that K+ deprivation of cerebellar
granule neurons induces cycloheximide-sensitive ICE-like protease
activity. A peptide inhibitor of ICE-like protease activity,
Ac-YVAD-chloromethylketone (Ac-YVAD-CMK), prevents
K+ deprivation-induced apoptosis. Further,
reactive oxygen species (ROS) are essential mediators of
K+ deprivation-induced apoptosis of cerebellar
granule neurons because neuronal death is also blocked by superoxide
dismutase, N-acetyl-L-cysteine, and
free radical spin traps. Using fluorescent assays, we show that ROS
production after K+ deprivation is blocked by
actinomycin D, cycloheximide, and Ac-YVAD-CMK, suggesting that ROS act
downstream of gene transcription, mRNA translation, and ICE activation.
Taken together, we show that new mRNA and protein synthesis, activation
of ICE-like proteases, and ROS production are sequential events in
K+ deprivation-induced apoptosis of cerebellar
granule neurons.
Key words:
apoptosis;
reactive oxygen species;
interleukin-1
converting enzyme;
ICE inhibition;
cerebellar granule neurons;
spin
traps;
superoxide dismutase
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