PMID- 12401327
OWN - NLM
STAT- MEDLINE
DCOM- 20030205
LR  - 20190712
IS  - 0306-4522 (Print)
IS  - 0306-4522 (Linking)
VI  - 115
IP  - 1
DP  - 2002
TI  - Immunohistochemical and biochemical assessment of caspase-3 activation and DNA 
      fragmentation following transient focal ischemia in the rat.
PG  - 125-36
AB  - In the present study, we evaluated the time-course of caspase-3 activation, and 
      the evolution of cell death following focal cerebral ischemia produced by 
      transient middle cerebral artery occlusion in rats. Ischemia-induced active 
      caspase-3 immunoreactivity in the striatum but not the cortex at 3 and 6 h time 
      points post-reperfusion. Furthermore, using a novel approach to visualize 
      enzymatic activity, deltaC-APP, a C-terminal cleavage product of APP generated by 
      caspase-3, was found to immunolocalize to the same areas as active caspase-3. 
      Double-labeling studies demonstrated co-localization of these two proteins at the 
      cellular level. Further double-labeling experiments revealed that active 
      caspase-3 was confined to neuronal cells which were still viable and thus 
      immunoreactive for NeuN. DNA fragmentation, assessed histologically by terminal 
      dUTP nick-end labeling (TUNEL), was observed in a small number of cells in the 
      striatum as early as 3 h, but only began to appear in the cortex by 6 h. DNA 
      fragmentation was progressive, and by 24 h post-reperfusion, large portions of 
      both the striatum and cortex showed TUNEL positive cells. However, 
      double-labeling of active caspase-3 with TUNEL showed only minimal 
      co-localization at all time-points. Thus, caspase-3 activation is an event that 
      appears to occur prior to DNA fragmentation. As a confirmation of the 
      histological TUNEL data, 24 h ischemia also induced the generation of nucleosome 
      fragments, evidenced by cell death enzyme-linked immunosorbent assay. Using a 
      novel ischemia-induced substrate cleavage biochemical approach, spectrin P120 
      fragment, a caspase-specific cleavage product of alpha II spectrin, a 
      cytoskeletal protein, was shown to be elevated by western blotting. Brain 
      concentrations of both nucleosomes and spectrin P120 correlate with the degree of 
      injury previously assessed by triphenyltetrazolium chloride staining and infarct 
      volume calculation. Together, our findings suggest a possible association between 
      caspase-3 activation and ischemic cell death following middle cerebral artery 
      occlusion brain injury.
FAU - Davoli, M A
AU  - Davoli MA
AD  - Merck Frosst Center for Therapeutic Research, P.O. Box 1005, Pointe 
      Claire-Dorval, QC, Canada H9R 4P8.
FAU - Fourtounis, J
AU  - Fourtounis J
FAU - Tam, J
AU  - Tam J
FAU - Xanthoudakis, S
AU  - Xanthoudakis S
FAU - Nicholson, D
AU  - Nicholson D
FAU - Robertson, G S
AU  - Robertson GS
FAU - Ng, G Y K
AU  - Ng GY
FAU - Xu, D
AU  - Xu D
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Neuroscience
JT  - Neuroscience
JID - 7605074
RN  - EC 3.4.22.- (Casp3 protein, rat)
RN  - EC 3.4.22.- (Caspase 3)
RN  - EC 3.4.22.- (Caspases)
SB  - IM
MH  - Animals
MH  - Caspase 3
MH  - Caspases/*analysis/*metabolism
MH  - Cell Death/genetics
MH  - *DNA Fragmentation/genetics
MH  - Enzyme Activation/genetics
MH  - Immunohistochemistry
MH  - In Situ Nick-End Labeling
MH  - Ischemic Attack, Transient/*enzymology/genetics
MH  - Male
MH  - Rats
MH  - Rats, Wistar
EDAT- 2002/10/29 04:00
MHDA- 2003/02/06 04:00
CRDT- 2002/10/29 04:00
PHST- 2002/10/29 04:00 [pubmed]
PHST- 2003/02/06 04:00 [medline]
PHST- 2002/10/29 04:00 [entrez]
AID - S0306452202003767 [pii]
AID - 10.1016/s0306-4522(02)00376-7 [doi]
PST - ppublish
SO  - Neuroscience. 2002;115(1):125-36. doi: 10.1016/s0306-4522(02)00376-7.