Neuronal apoptosis inhibitory protein expression after traumatic brain injury in the mouse.
Apoptosis of brain cells is triggered by traumatic brain injury (TBI) and is blocked by caspase inhibitors. The neuronal apoptosis inhibitor protein (NAIP), which has been shown to inhibit apoptosis by both caspase-dependant and caspase-independent mechanisms, is neuroprotective in rat models of cerebral ischemia and axotomy. In order to gain a better appreciation of CNS apoptosis following head injury in general and the possible involvement of NAIP specifically, we have configured a mouse model of TBI. In addition to demonstrating apoptosis, the spatiotemporal expression or levels of a number of proteins with apoptosis modulating effects have been determined. Apoptosis of neurons and oligodendrocytes following TBI was observed in brain sections which were triple-stained with in situ end labeling, bisbenzimide and immunofluorescent stain for neuron specific nuclear protein and myelin-associated glycoprotein, respectively. Further evidence for apoptosis following TBI in this model was obtained in brain samples using ligation-mediated PCR amplification of DNA fragments and gel electrophoresis. The temporal profile of apoptosis was similar to the temporal profile of microglial activation determined by CD11b staining and TNFa expression induced by TBI. NAIP staining in sections of cerebral cortex and subcortical white matter increased at 6 h and decreased towards control levels at 24 h post-TBI. Temporal changes in the expression of NAIP were also observed using Western blot analysis of brain samples removed from injured cortex and sub-cortical white matter. At the time that NAIP expression decreased markedly (24 h post-TBI), procaspase-3 levels also decreased, PARP cleavage increased, and the highest levels of apoptosis were observed. These findings have implications in our understanding of traumatically induced programmed cell death and may be useful in the configuration of therapies for this common injury state.
Hutchison
J S
JS
Department of Pediatrics, Faculty of Medicine, University of Ottawa, Ontario, Canada. hutchison@cheo.on.ca
Derrane
R E
RE
Johnston
D L
DL
Gendron
N
N
Barnes
D
D
Fliss
H
H
King
W J
WJ
Rasquinha
I
I
MacManus
J
J
Robertson
G S
GS
MacKenzie
A E
AE
D000818:Animals; D017209:Apoptosis / Q000502:physiology; D001930:Brain Injuries / Q000378:metabolism* / Q000473:pathology; D053148:Caspase 3; D020169:Caspases / Q000096:biosynthesis; D002540:Cerebral Cortex / Q000378:metabolism; D004195:Disease Models, Animal; D004792:Enzyme Precursors / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000172:deficiency / Q000235:genetics; D051616:Neuronal Apoptosis-Inhibitory Protein; D009474:Neurons / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
D004792:Enzyme Precursors; C496801:Naip1 protein, mouse; D009419:Nerve Tissue Proteins; D051616:Neuronal Apoptosis-Inhibitory Protein; D014409:Tumor Necrosis Factor-alpha; C505409:Casp3 protein, mouse; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases