Abstract
A major consequence of traumatic brain injury (TBI) is the rapid proteolytic degradation of structural cytoskeletal proteins. This process is largely reflected by the interruption of axonal transport as a result of extensive axonal injury leading to neuronal cell injury. Previous work from our group has described the extensive degradation of the axonally enriched cytoskeletal αII-spectrin protein which results in molecular signature breakdown products (BDPs) indicative of injury mechanisms and to specific protease activation both in vitro and in vivo. In the current study, we investigated the integrity of βII-spectrin protein and its proteolytic profile both in primary rat cerebrocortical cell culture under apoptotic, necrotic, and excitotoxic challenge and extended to in vivo rat model of experimental TBI (controlled cortical impact model). Interestingly, our results revealed that the intact 260-kDa βII-spectrin is degraded into major fragments (βII-spectrin breakdown products (βsBDPs)) of 110, 108, 85, and 80 kDa in rat brain (hippocampus and cortex) 48 h post-injury. These βsBDP profiles were further characterized and compared to an in vitro βII-spectrin fragmentation pattern of naive rat cortex lysate digested by calpain-2 and caspase-3. Results revealed that βII-spectrin was degraded into major fragments of 110/85 kDa by calpain-2 activation and 108/80 kDa by caspase-3 activation. These data strongly support the hypothesis that in vivo activation of multiple protease system induces structural protein proteolysis involving βII-spectrin proteolysis via a specific calpain and/or caspase-mediated pathway resulting in a signature, protease-specific βsBDPs that are dependent upon the type of neural injury mechanism. This work extends on previous published work that discusses the interplay spectrin family (αII-spectrin and βII-spectrin) and their susceptibility to protease proteolysis and their implication to neuronal cell death mechanisms.
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27 September 2017
An erratum to this article has been published.
Abbreviations
- TBI:
-
Traumatic brain injury
- αII-SBDPs:
-
αII-Spectrin breakdown products
- βsBDPs:
-
βII-Spectrin breakdown products
- BDPs:
-
Breakdown products
- CCI:
-
Controlled cortical impact
- EDTA:
-
Ethylenediaminetetraacetic acid
- MTX:
-
Maitotoxin
- NMDA:
-
N-Methyl-d-aspartate
- STS:
-
Staurosporine
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Acknowledgments
We would like to thank Dr. Hussam Jourdi for his critical and thorough discussion. Special thanks to Mr. Danny Johnson for his technical support in animal surgeries and tissue collection. This work was supported by the National Institutes of Health grants R01 NS049175-01 and R01 NS052831-01 and the Department of Defense grant DAMD17-03-1-0066. KKW and RLH hold equity in Banyan Biomarkers, Inc., a company commercializing technology of detecting brain injury biomarkers. RLH and OG are employees at Banyan Biomarkers Inc.
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An erratum to this article is available at https://doi.org/10.1007/s12035-017-0764-3.
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Kobeissy, F.H., Liu, M.C., Yang, Z. et al. Degradation of βII-Spectrin Protein by Calpain-2 and Caspase-3 Under Neurotoxic and Traumatic Brain Injury Conditions. Mol Neurobiol 52, 696–709 (2015). https://doi.org/10.1007/s12035-014-8898-z
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DOI: https://doi.org/10.1007/s12035-014-8898-z