Novel Differential Neuroproteomics Analysis of Traumatic Brain Injury in Rats

Abstract

Approximately two million traumatic brain injury (TBI) incidents occur annually in the United States, yet there are no specific therapeutic treatments. The absence of brain injury diagnostic endpoints was identified as a significant roadblock to TBI therapeutic development. To this end, our laboratory has studied mechanisms of cellular injury for biomarker discovery and possible therapeutic strategies. In this study, pooled naïve and injured cortical samples (48 h postinjury; rat controlled cortical impact model) were processed and analyzed using a differential neuroproteomics platform. Protein separation was performed using combined cation/anion exchange chromatography-PAGE. Differential proteins were then trypsinized and analyzed with reversed-phase LC-MSMS for protein identification and quantitative confirmation. The results included 59 differential protein components of which 21 decreased and 38 increased in abundance after TBI. Proteins with decreased abundance included collapsin response mediator protein 2 (CRMP-2), glyceraldehyde-3-phosphate dehydrogenase, microtubule-associated proteins MAP2A/2B, and hexokinase. Conversely C-reactive protein, transferrin, and breakdown products of CRMP-2, synaptotagmin, and ␣II-spectrin were found to be elevated after TBI. Differential changes in the above mentioned proteins were confirmed by quantitative immunoblotting. Results from this work provide insight into mechanisms of traumatic brain injury and yield putative biochemical markers to potentially facilitate patient management by monitoring the severity, progression, and treatment of injury.

Effects of cathepsins B and L inhibition on postischemic protein alterations in the brain

“…A number of proteomic studies have been published in recent years that investigated alterations in protein expression relevant to acute and chronic CNS diseases [30][31][32][33][34][35][36]. However, out of the two stroke-related proteomics studies published, there was only one that examined protein alterations in the brain after experimental stroke [17].…”

Stretching the limits: Stem cells in regeneration science

“…For example, 48 hours after closed head injury, cortical tissue collected from adult rats has revealed a breakdown of collapsin response mediator protein-2 (Kobeissy et al, 2006). In Caenorhabditis elegans, this molecule is required for proper axon morphology and pathfinding (Hedgecock et al, 1985;Siddiqui and Culotti, 1991).…”

Biomarkers of Traumatic Brain Injury in the Geriatric Population

“…However, the emergence of new technologies such as cerebral microdialysis, advances in brain imaging, and identification of TBI-specific biological markers may expand our ability to appropriately stratify and manage TBI. Recently, the pursuit of neuronal biomarker panels has gained momentum for the potential to provide real-time monitoring of the severity and progression of injury at the cellular level [31]. Knowledge at this level of detail may translate into injury-specific TBI stratification and titration and optimization of treatment modalities, and may guide development of cutting edge therapeutics [32].…”