While caspase-3 has been widely implicated in apoptosis, rapid progress in understanding mechanisms of apoptotic cell death is difficult in in vivo models of traumatic brain injury (TBI). Traumatic brain injury is multifactorial, primarily including tissue deformation and secondary hypoxia and ischemia, all of which could result in release of excitotoxic levels of glutamate. We have developed a battery of in vitro systems to study separately three major pathological components of traumatic brain injury: (1) biomechanical stretch injury, (2) ischemia and/or hypoxia (oxygen and/or glucose deprivation)and (3) glutamate-induced excitotoxicity.
Aim 1 will characterize caspase-3 and calpain activation in these in vitro models and examine the contribution of these two families of cysteine proteases to archetypal apoptotic and necrotic cell death phenotypes. Researchers have described at least two major apoptotic cell death pathways in mammals regulating cspase-3 activation: (1) a mitochondrial pathway requiring cspase-9 activation and (2) a receptor mediated signal transduction pathway principally involving tumor necrosis factor (TNF-a) and requiring caspase-8/10 activation. Both of these pathways could regulate caspase-3 activation.
AIM 2 will examine the mitochondrial cell death pathway since this pathway is well characterized in non- neuronal systems, the requisite cellular machinery is present in neurons and data implicate a mitochondrial contribution to neuronal injury and apoptotic cell death.
Aim 3 will examine TNF-a receptor coupled pathway since laboratory and clinical studies suggest that traumatic brain injury increases levels of TNF-a that could contribute to apoptotic neuronal death.
Aim 4 will test the generality of in vitro mechanisms of cell death characterized in Aims 1-3 in a clinically relevant in vivo model of traumatic brain injury. Our general hypothesis is that caspase-3 activation by casepase-9 and/or caspase 8/10 is an important common mediator of apoptotic cell death following CNS injury. Calpain activation contributes primarily to necrosis and, to a lesser extent, apoptosis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS039091-02
Application #
6394214
Study Section
Special Emphasis Panel (ZRG1-BDCN-3 (01))
Program Officer
Michel, Mary E
Project Start
2000-09-15
Project End
2004-08-31
Budget Start
2001-09-01
Budget End
2002-08-31
Support Year
2
Fiscal Year
2001
Total Cost
$344,546
Indirect Cost
Name
University of Florida
Department
Neurosciences
Type
Schools of Medicine
DUNS #
073130411
City
Gainesville
State
FL
Country
United States
Zip Code
32611
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Johnson, Erik A; Svetlov, Stanislav I; Wang, Kevin K W et al. (2005) Cell-specific DNA fragmentation may be attenuated by a survivin-dependent mechanism after traumatic brain injury in rats. Exp Brain Res 167:17-26
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Pineda, Jose A; Wang, Kevin K; Hayes, Ronald L (2004) Biomarkers of proteolytic damage following traumatic brain injury. Brain Pathol 14:202-9
Larner, Stephen F; Hayes, Ronald L; McKinsey, Deborah M et al. (2004) Increased expression and processing of caspase-12 after traumatic brain injury in rats. J Neurochem 88:78-90
Shepherd, Timothy M; Thelwall, Peter E; Blackband, Stephen J et al. (2003) Diffusion magnetic resonance imaging study of a rat hippocampal slice model for acute brain injury. J Cereb Blood Flow Metab 23:1461-70
Tolentino, Paul J; DeFord, S Michelle; Notterpek, Lucia et al. (2002) Up-regulation of tissue-type transglutaminase after traumatic brain injury. J Neurochem 80:579-88

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