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Traumatic brain injury induces upregulation of cytosol-localized procaspase-3 and mitochondrial-localized activated caspase-3


, : Traumatic brain injury induces upregulation of cytosol-localized procaspase-3 and mitochondrial-localized activated caspase-3. Society for Neuroscience Abstracts 26(1-2): Abstract No -863 2

The neuropathalogic sequalae of traumatic brain injury (TBI) includes regionally distinct patterns of neuronal apoptosis. Activation of caspase-3, an effector caspase with known cytosolic and mitochondrial distributions, is a critical mechanism underlying apoptotic cell death. We characterized the subcellular distribution and time-course of caspase-3 activation following brain injury in the rat, using a polyclonal antibody (Ab 206) that recognizes both procaspase- and activated caspase-3. Anesthetized adult male Sprague-Dawley rats (n=8) were subjected to lateral fluid percussion (FP) brain injury of moderate severity (2.4-2.7 atm), while sham controls (n=2) were surgically prepared but did not receive brain injury. Animals were sacrificed by decapitation at 2 h (n=2), 6 h (n=3), and 24 h (n=3), their injured cortices dissected and homogenized, and the resulting tissue subfractionated into cytosolic and mitochondrial fractions. The fractions were then subjected to electrophoresis and subsequent Western blotting, probing for the procaspase- and activated caspase-3. Western blots revealed a dramatic, time-dependent increase in expression of the 32-kD procaspase-3 at 2, 6, and 24 h in cytosolic fractions following TBI. The 17-kD activated caspase-3 fragment was not detected in cytosolic fractions. In the mitochondrial fractions, Western blots revealed an increase in expression of the 17-kD activated caspase-3 fragment at 2 h and 6 h post-injury. These results suggest that caspase-3 activation may be indicative of a unique mechanism for posttraumatic mitochondrial dysfunction.

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