Proteomic analysis of injured spinal cord tissue proteins using 2-DE and MALDI-TOF MS

Abstract

Spinal cord injury (SCI) induces a progressive pathophysiology affecting cell survival and neurological integrity via complex and evolving molecular cascades whose interrelationships are not fully understood. Acute injury to the spinal cord undergoes sequential pathological change including hemorrhage, edema, axonal and neuronal necrosis, and demyelination. In the present study, we aimed to establish the proteomic profiles and characterization of the total protein expressed in traumatic injured spinal cord tissue by using 2-DE and matrix assisted laser desorption/ionization-TOF MS (MALDI-TOF MS). We performed proteomic analysis using 2-DE and MS to describe total proteins and differential proteins expression between normal and traumatic injured spinal cord tissues. The study discovered 947 total proteins and analyzed 219 and 270 proteins from normal and injured tissue, respectively. After 24 h of traumatic damage induction, the injured spinal cord tissue up-regulated over 39 proteins including neurofilament light chain, annexin 5, heat shock protein, tubulin beta, peripherin, glial fibrillary acidic protein delta, peroxiredoxin 2, and apolipoprotein A. Twenty-one proteins showed reduction. The majority of the modulated proteins belonged to the 13 functional categories. Proteins that were identified with neural functional category in injured tissue were considered most likely to be involved in wound healing response coupled with neurogenesis and gliogenesis.

Involvement of Acidic Fibroblast Growth Factor in Spinal Cord Injury Repair Processes Revealed by a Proteomics Approach

“…It is worth noting that from our 2D PAGE analysis we could only detect HPT in the contusive SCI group but not in the sham group. Moreover the up-regulation of albumin and transferrin was in agreement with a previous study using proteomics analysis of transection-injured spinal cord tissue of rats (34,35). Most importantly, the up-regulation of these proteins was decreased in the aFGF-treated spinal cord tissue after SCI.…”

“…To date, there have been only two reports involving the study of protein profiles of spinal cord injury using a proteomics approach (34,35). However, Ding et al (34) did not dissect the injured spinal segments from the uninjured neighboring tissues, and therefore it is unknown whether the differentially expressed proteins detected in this study were localized in injured spinal region or elsewhere.…”

“…Over the past few years, a large number of studies have been carried out to investigate the pathology and regenerative processes that take place in the injured spinal cord using both transcriptional and translational level strategies involving microarrays and proteomics (9,34,35), respectively. In addition to these previous reports, in this work, we used comparative proteomics techniques to screen for differences in protein expression levels between sham, contusion-injured, and contusion-injured aFGF-treated spinal cord tissue in rats.…”

Dynamic proteome analysis of spinal cord injury after ischemia–reperfusion in rabbits by two-dimensional difference gel electrophoresis

“…Glial fibrillary acidic protein (a class-III intermediate filament protein) was also downregulated at this time point, although a previous study demonstrated glial fibrillary acidic protein upregulation in SCI induced by laminectomy. 4 NEFM, another neurofilament protein, was distributed at six spots with both up-and downregulated patterns. A previous study reported that NEFM expression is influenced strongly in oxidative stress, and estradiol may protect neurons by inducing NEFM expression.…”

“…1 Reactive oxidation species have been implicated in this pathogenesis 2 and recent proteomics studies of primary stage SCI have identified some important molecules. 3,4 However, the exact mechanism of ischemia-reperfusion-induced SCI remains unclear and the protein differential level of the second stage of injury is seldom reported. High-throughput proteomics tools promise discovery of molecules involved in the pathological processes of ischemia-reperfusion injury.…”