Evidence for activation of caspase-3-like protease in excitotoxin- and hypoxia/hypoglycemia-injured neurons

J Neurochem. 1998 Jul;71(1):186-95. doi: 10.1046/j.1471-4159.1998.71010186.x.


Caspase activation has been shown to be a critical step in several models of neuronal apoptosis such as staurosporine treatment of human neuroblastoma SH-SY5Y cells and potassium deprivation of rat cerebellar granule neurons. One common event is the appearance of caspase-mediated 120-kDa nonerythroid alpha-spectrin breakdown product (SBDP120). Second, inhibitors of the caspase family are effective blockers of such neuronal death. In this study, we report the appearance of caspase-mediated SBDP120 in excitotoxin-challenged fetal rat cerebrocortical neurons [N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, and kainate] and rat cerebellar granule neurons (NMDA and kainate). A general caspase inhibitor, carbobenzoxy-Asp-CH2OC(O)-2,6-dichlorobenzene (Z-D-DCB), blocked the formation of SBDP120 under these conditions and attenuated the observed NMDA-induced lactate dehydrogenase (LDH) release in both cell types. Furthermore, hydrolytic activity toward a caspase-3-preferred synthetic peptide substrate, acetyl-DEVD-7-amido-4-methylcoumarin, was significantly elevated in NMDA-treated granule neurons. Lastly, oxygen-glucose deprivation (OGD)-challenged cerebrocortical cultures also showed the appearance of SBDP120. Again, Z-D-DCB blocked the SBDP120 formation as well as attenuated the LDH release from the OGD-challenged neurons. Taken together, the presence of caspase-specific SBDP120 and the neuroprotective effects of Z-D-DCB strongly suggest that caspase activation contributes at least in part to excitotoxin- and OGD-induced neuronal death.

MeSH terms

  • Animals
  • Caspase 3
  • Caspases*
  • Cell Death / drug effects
  • Cell Death / physiology
  • Cell Hypoxia / physiology
  • Cells, Cultured
  • Cerebellum / cytology
  • Cerebral Cortex / cytology
  • Cysteine Endopeptidases / metabolism*
  • Enzyme Precursors / metabolism
  • Excitatory Amino Acid Agonists / pharmacology
  • Fetus / cytology
  • Glucose / pharmacology
  • Hypoglycemia / metabolism
  • Kainic Acid / pharmacology
  • N-Methylaspartate / pharmacology
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / enzymology*
  • Neurotoxins / pharmacology*
  • Oligopeptides / pharmacology
  • Peptide Fragments / analysis
  • Protease Inhibitors / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Spectrin / metabolism


  • Enzyme Precursors
  • Excitatory Amino Acid Agonists
  • Neurotoxins
  • Oligopeptides
  • Peptide Fragments
  • Protease Inhibitors
  • benzyloxycarbonyl-Glu-Val-Asp-CH2OC(O)-2,6-dichlorobenzene
  • Spectrin
  • N-Methylaspartate
  • CASP3 protein, human
  • Casp3 protein, rat
  • Caspase 3
  • Caspases
  • Cysteine Endopeptidases
  • Glucose
  • Kainic Acid