Proteome of synaptosome-associated proteins in spinal cord dorsal horn after peripheral nerve injury.
Peripheral nerve injury may lead to neuroadaptive changes of cellular signals in spinal cord that are thought to contribute to central mechanisms underlying neuropathic pain. Here we used a 2-DE-based proteomic technique to determine the global expression changes of synaptosome-associated proteins in spinal cord dorsal horn after unilateral fifth spinal nerve injury (SNI). The fifth lumbar dorsal horns ipsilateral to SNI or sham surgery were harvested on day 14 post-surgery, and the total soluble and synaptosomal fractions were isolated. The proteins derived from the synaptosomal fraction were resolved by 2-DE. We identified 27 proteins that displayed different expression levels after SNI, including proteins involved in transmission and modulation of noxious information, cellular metabolism, membrane receptor trafficking, oxidative stress, apoptosis, and degeneration. Six of the 27 proteins were chosen randomly and further validated in the synaptosomal fraction by Western blot analysis. Unexpectedly, Western blot analysis showed that only one protein in the total soluble fraction exhibited a significant expression change after SNI. The data indicate that peripheral nerve injury changes not only protein expression but also protein subcellular distribution in dorsal horn cells. These changes might participate in the central mechanism that underlies the maintenance of neuropathic pain.
Singh
Om V
OV
Department of Pediatrics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Yaster
Myron
M
Xu
Ji-Tian
JT
Guan
Yun
Y
Guan
Xiaowei
X
Dharmarajan
Arun M
AM
Raja
Srinivasa N
SN
Zeitlin
Pamela L
PL
Tao
Yuan-Xiang
YX
D000818:Animals; D017209:Apoptosis; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D008297:Male; D013058:Mass Spectrometry; D023421:Models, Animal; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress; D020671:Posterior Horn Cells / Q000737:chemistry / Q000378:metabolism*; D020543:Proteome / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011956:Receptors, Cell Surface / Q000378:metabolism; D013119:Spinal Cord Injuries / Q000235:genetics / Q000378:metabolism*; D013574:Synaptosomes / Q000737:chemistry* / Q000378:metabolism*
D009419:Nerve Tissue Proteins; D020543:Proteome; D011956:Receptors, Cell Surface