A gel-based proteomic comparison of human cerebrospinal fluid between inflicted and non-inflicted pediatric traumatic brain injury.
Traumatic brain injury (TBI) is the most common cause of traumatic death in infancy, and inflicted TBI (iTBI) is the predominant cause. Like other central nervous system pathologies, TBI changes the composition of cerebrospinal fluid (CSF), which may represent a unique clinical window on brain pathophysiology. Proteomic analysis, including two-dimensional (2-D) difference in gel electrophoresis (DIGE) combined with mass spectrometry (MS), was used to compare the CSF protein profile of two pooled samples from pediatric iTBI (n = 13) and non-inflicted TBI (nTBI; n = 13) patients with severe injury. CSF proteins from iTBI and nTBI were fluorescently labeled in triplicate using different fluorescent Cy dyes and separated by 2-D gel electrophoresis. Approximately 250 protein spots were found in CSF, with 90% between-gel reproducibility of the 2-D gel. Following in-gel digestion, the tryptic peptides were analyzed by MS for protein identification. The acute phase reactant, haptoglobin (HP) isoforms, showed an approximate fourfold increase in nTBI versus iTBI. In contrast, the levels of prostaglandin D(2) synthase (PGDS) and cystatin C (CC) were 12-fold and sevenfold higher in iTBI versus nTBI, respectively. The changes of HP, PGDS, and CC were confirmed by Western blot. These initial results with conventional gel-based proteomics show new protein changes that may ultimately help to understand pathophysiological differences between iTBI and nTBI.
Gao
Wei-Min
WM
Department of Critical Care Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA 15260, USA.
Chadha
Mandeep S
MS
Berger
Rachel P
RP
Omenn
Gilbert S
GS
Allen
David L
DL
Pisano
Michael
M
Adelson
P David
PD
Clark
Robert S B
RS
Jenkins
Larry W
LW
Kochanek
Patrick M
PM
D001930:Brain Injuries / Q000134:cerebrospinal fluid* / Q000175:diagnosis; D002555:Cerebrospinal Fluid / Q000737:chemistry*; D002649:Child Abuse / Q000175:diagnosis*; D002675:Child, Preschool; D055316:Cystatin C; D015891:Cystatins / Q000134:cerebrospinal fluid; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D008401:Gas Chromatography-Mass Spectrometry; D006242:Haptoglobins / Q000134:cerebrospinal fluid; D006801:Humans; D015151:Immunoblotting; D007223:Infant; D019746:Intramolecular Oxidoreductases / Q000134:cerebrospinal fluid; D054834:Lipocalins; D008297:Male; D013058:Mass Spectrometry; D020543:Proteome / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
C525030:CST3 protein, human; D055316:Cystatin C; D015891:Cystatins; D006242:Haptoglobins; D054834:Lipocalins; D020543:Proteome; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase