Neurocritical Care

, Volume 12, Issue 3, pp 324–336 | Cite as

Metabolic Crisis After Traumatic Brain Injury is Associated with a Novel Microdialysis Proteome

  • R. Lakshmanan
  • J. A. Loo
  • T. Drake
  • J. Leblanc
  • A. J. Ytterberg
  • D. L. McArthur
  • M. Etchepare
  • P. M. VespaEmail author
Original Article



To examine if the metabolic distress after traumatic brain injury (TBI) is associated with a unique proteome.


Patients with severe TBI prospectively underwent cerebral microdialysis for the initial 96 h after injury. Hourly sampling of metabolism was performed and patients were categorized as having normal or abnormal metabolism as evidenced by the lactate/pyruvate ratio (LPR) threshold of 40. The microdialysate was frozen for proteomic batch processing retrospectively. We employed two different routes of proteomic techniques utilizing mass spectrometry (MS) and categorized as diagnostic and biomarker identification approaches. The diagnostic approach was aimed at finding a signature of MS peaks which can differentiate these two groups. We did this by enriching for intact peptides followed by MALDI-MS analysis. For the biomarker identification approach, we applied classical bottom-up (trypsin digestion followed by LC-MS/MS) proteomic methodologies.


Five patients were studied, 3 of whom had abnormal metabolism and 2 who had normal metabolism. By comparison, the abnormal group had higher LPR (1609 ± 3691 vs. 15.5 ± 6.8, P < 0.001), higher glutamate (157 ± 84 vs. 1.8 ± 1.4 μM, P < 0.001), and lower glucose (0.27 ± 0.35 vs. 1.8 ± 1.1 mmol/l, P < 0.001). The abnormal group demonstrated 13 unique proteins as compared with the normal group in the microdialysate. These proteins consisted of cytoarchitectural proteins, as well as blood breakdown proteins, and a few mitochondrial proteins. A unique as yet to be characterized peptide was found at m/z (mass/charge) 4733.5, which may represent a novel biomarker of metabolic distress.


Metabolic distress after TBI is associated with a differential proteome that indicates cellular destruction during the acute phase of illness. This suggests that metabolic distress has immediate cellular consequences after TBI.


Traumatic brain injury Microdialysis Proteomics Metabolic crisis Biomarkers 


  1. 1.
    Marcoux J, McArthur DA, Miller C, Glenn TC, Villablanca P, Martin NA, et al. Persistent metabolic crisis as measured by elevated cerebral microdialysis lactate-pyruvate ratio predicts chronic frontal lobe brain atrophy after traumatic brain injury. Crit Care Med. 2008;36:2871–7.CrossRefPubMedGoogle Scholar
  2. 2.
    Bullock R, Zauner A, Woodward JJ, Myseros J, Choi SC, Ward JD, et al. Factors affecting excitatory amino acid release following human head injury. J Neurosurg. 1998;89:507–18.CrossRefPubMedGoogle Scholar
  3. 3.
    Vespa M, Prins M, Ronne-Engstrom E, Caron M, Shalmon E, Hovda DA, et al. Increase in extracellular glutamate caused by reduced cerebral perfusion presure and seizures after human traumatic brain injury: a microdialysis study. J Neurosurg. 1998;89:971–82.CrossRefPubMedGoogle Scholar
  4. 4.
    Hillered L, Vespa PM, Hovda DA. Translational neurochemical research in acute human brain injury: the current status and potential future for cerebral microdialysis. J Neurotrauma. 2005;22:3–41.CrossRefPubMedGoogle Scholar
  5. 5.
    Pineda JA, Wang KK, Hayes RL. Biomarkers of proteolytic damage following traumatic brain injury. Brain Pathol. 2004;14:202–9.PubMedCrossRefGoogle Scholar
  6. 6.
    Berger RP. The use of serum biomarkers to predict outcome after traumatic brain injury in adults and children. J Head Trauma Rehabil. 2006;21:315–33.CrossRefPubMedGoogle Scholar
  7. 7.
    Haqqani AS, Hutchison JS, Ward R, Stanimirovic DB. Biomarkers and diagnosis; protein biomarkers in serum of pediatric patients with severe traumatic brain injury identified by ICAT-LC-MS/MS. J Neurotrauma. 2007;24:54–74.CrossRefPubMedGoogle Scholar
  8. 8.
    Gao WM, Chadha MS, Berger RP, Omenn GS, Allen DL, Pisano M, et al. A gel-based proteomic comparison of human cerebrospinal fluid between inflicted and non-inflicted pediatric traumatic brain injury. J Neurotrauma. 2007;24:43–53.CrossRefPubMedGoogle Scholar
  9. 9.
    Gobom J, Nordhoff E, Mirgorodskaya E, Ekman R, Roepstorff P. Sample purification and preparation technique based on nano-scale reversed-phase columns for the sensitive analysis of complex peptide mixtures by matrix-assisted laser desorption/ionization mass spectrometry. J Mass Spectrom. 1999;34(2):105–16.CrossRefPubMedGoogle Scholar
  10. 10.
    Larsen MR, Sørensen GL, Fey SJ, Larsen PM, Roepstorff P. Phospho-proteomics: evaluation of the use of enzymatic de-phosphorylation and differential mass spectrometric peptide mass mapping for site specific phosphorylation assignment in proteins separated by gel electrophoresis. Proteomics. 2001;1(2):223–38.CrossRefPubMedGoogle Scholar
  11. 11.
    Zuberovic A, Wetterhall M, Hanrieder J, Bergquist J. CE MALDI-TOF/TOF MS for multiplexed quantification of proteins in human ventricular cerebrospinal fluid. Electrophoresis. 2009;30:1836–43.CrossRefPubMedGoogle Scholar
  12. 12.
    Hanrieder J, Wetterhall M, Enblad P, Hillered L, Bergquist J. Temporally resolved differential proteomic analysis of human ventricular CSF for monitoring traumatic brain injury biomarker candidates. J Neurosci Methods. 2009;177:469–78.CrossRefPubMedGoogle Scholar
  13. 13.
    Lumpkins KM, Bochicchio GV, Keledjian K, Simard JM, McCunn M, Scalea T. Glial fibrillary acidic protein is highly correlated with brain injury. J Trauma. 2008;65:778–82. discussion 782–774.CrossRefPubMedGoogle Scholar
  14. 14.
    Zemlan FP, Jauch EC, Mulchahey JJ, Gabbita SP, Rosenberg WS, Speciale SG, et al. C-tau biomarker of neuronal damage in severe brain injured patients: association with elevated intracranial pressure and clinical outcome. Brain Res. 2002;947:131–9.CrossRefPubMedGoogle Scholar
  15. 15.
    Afinowi R, Tisdall M, Keir G, Smith M, Kitchen N, Petzold A. Improving the recovery of S100B protein in cerebral microdialysis: implications for multimodal monitoring in neurocritical care. J Neurosci Methods. 2009;181:95–9.CrossRefPubMedGoogle Scholar
  16. 16.
    Maurer MH, Berger C, Wolf M, Futterer CD, Feldmann RE Jr, Schwab S, et al. The proteome of human brain microdialysate. Proteome Sci. 2003;1:7.CrossRefPubMedGoogle Scholar
  17. 17.
    Brody DL, Magnoni S, Schwetye K, Spinner ML, Esparza TL, Stocchetti N, et al. Science. 2008;321:1221–4.CrossRefPubMedGoogle Scholar
  18. 18.
    Marklund N, Blennow K, Zetterberg H, Ronne-Engstrom E, Enblad P, Hillered L. Monitoring of brain interstitial total tau and beta amyloid proteins by microdialysis in patients with traumatic brain injury. J Neurosurgery. 2009;110:1227–37.CrossRefGoogle Scholar
  19. 19.
    Cassimeris L. The oncoprotein 18/stathmin family of microtubule destabilizers. Curr Opin Cell Biol. 2002;14(1):18–24.CrossRefPubMedGoogle Scholar
  20. 20.
    Ottens AK, Kobeissy F, Golden E, Ahang A, Haskins W, Chen S, et al. Neuroproteomics in neurotrauma. Mass Spec Rev. 2006;25:380–408.CrossRefGoogle Scholar
  21. 21.
    Yang X, Yang S, Wang J, Zhang X, Wang C, Hong G. Expressive proteomics profile changes of injured human brain cortex due to acute brain trauma. Brain Injury. 2009;23:830–40.CrossRefPubMedGoogle Scholar
  22. 22.
    Vespa P, Bergsneider M, Hattori N, Wu HM, Huang SC, Martin NA, et al. Metabolic crisis without brain ischemia is common after traumatic brain injury: a combined microdialysis and positron emission tomography study. J Cereb Blood Flow Metab. 2005;25:763–74.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • R. Lakshmanan
    • 1
  • J. A. Loo
    • 1
  • T. Drake
    • 2
  • J. Leblanc
    • 2
  • A. J. Ytterberg
    • 1
  • D. L. McArthur
    • 3
  • M. Etchepare
    • 3
  • P. M. Vespa
    • 3
    Email author
  1. 1.UCLA Department of Chemistry and BiochemistryDavid Geffen School of Medicine at UCLALos AngelesUSA
  2. 2.UCLA Department of Pathology and Laboratory MedicineDavid Geffen School of Medicine at UCLALos AngelesUSA
  3. 3.UCLA Department of Neurosurgery and NeurologyDavid Geffen School of Medicine at UCLA, Ronald Reagan UCLA Medical CenterLos AngelesUSA

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