Cytochrome c release and caspase-3 activation during colchicine-induced apoptosis of cerebellar granule cells

Eur J Neurosci. 1999 Mar;11(3):1067-72. doi: 10.1046/j.1460-9568.1999.00512.x.


The microtubule-disrupting agent colchicine is known to be neurotoxic toward certain neuronal populations including cerebellar granule cells (CGCs). In this study we investigated the involvement of cytochrome c release and caspase-3 activation during colchicine-induced CGC apoptosis. Treatment of rat CGCs with 1 micrometer colchicine (for up to 24 h) caused high molecular weight DNA fragmentation and nuclear condensation. An involvement of group II caspases (which includes caspase-3) was demonstrated by the proteolytic degradation of poly(ADP-ribose) polymerase (PARP) after 18 h exposure to colchicine. Colchicine induced a time-dependent increase in Ac-Asp-Glu-Val-Asp-alpha-(4-methyl-coumaryl-7-amide) (DEVD-MCA) cleavage activity in CGCs, which was blocked with a specific, peptide-based, aldehyde inhibitor of group II caspases, i. e. DEVD-CHO. We also observed a time-dependent proteolysis of caspase-3 as judged by the appearance of p17 which is one of the subunits of active caspase-3. Activation of caspase-3 during colchicine-induced apoptosis may be mediated by cytochrome c since there was a close correlation between the time courses of cytochrome c release from the mitochondria and of caspase-3 activation. Furthermore, colchicine-induced apoptosis, as assessed by propidium iodide visualization of the nuclei, could be blocked by the caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp (O-methyl) fluoromethyl ketone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Animals
  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Caspase 3
  • Caspases / metabolism*
  • Cells, Cultured
  • Cerebellum / cytology
  • Colchicine / pharmacology
  • Coumarins / pharmacology
  • Cysteine Proteinase Inhibitors / pharmacology
  • Cytochrome c Group / metabolism*
  • Enzyme Precursors / metabolism
  • Mitochondria / enzymology
  • Neurons / cytology*
  • Neurons / enzymology*
  • Oligopeptides / pharmacology
  • Poly(ADP-ribose) Polymerases / metabolism
  • Rats
  • Rats, Sprague-Dawley
  • Time Factors


  • Amino Acid Chloromethyl Ketones
  • Coumarins
  • Cysteine Proteinase Inhibitors
  • Cytochrome c Group
  • Enzyme Precursors
  • Oligopeptides
  • aspartyl-glutamyl-valyl-aspartal
  • benzyloxycarbonyl-aspartyl-glutamyl-valyl-aspartyl-7-amino-4-trifluoromethylcoumarin
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • Poly(ADP-ribose) Polymerases
  • Casp3 protein, rat
  • Caspase 3
  • Caspases
  • Colchicine