Abstract
Nerve injury-induced neuropathic pain is a major public health problem worldwide. Current treatment for neuropathic pain has had limited success because the mechanisms that underlie the induction and maintenance of neuropathic pain are incompletely understood. However, recent advances in proteomics may allow us to uncover complicated biological mechanisms that occur under neuropathic pain conditions. Here, we introduce a combined approach of two-dimensional gel electrophoresis (2-DE) with mass spectrometry (MS) to identify the expression changes in synaptosome-associated proteins in spinal cord dorsal horn after unilateral fifth spinal nerve injury. In 2-DE, a set of highly abundant synaptic proteins with a pI range of 4–7 are separated and compared by size fractionation (25–100 kDa). Then, the differentially expressed proteins are identified and validated by MS, and their potential involvement in physiological and pathological processes is searched. Thus, proteomic analysis can provide expression profiles of synaptic proteins and their posttranslational modifications in cells, tissues, and organs of the nervous system under neuropathic pain conditions.
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References
Campbell, J. N. and Meyer, R. A. (2006) Mechanisms of neuropathic pain. Neuron 52, 77–92.
Latremoliere, A. and Woolf, C. J. (2009) Central sensitization: a generator of pain hypersensitivity by central neural plasticity. J. Pain 10, 895–926.
Price, T. J. and Geranton, S. M. (2009) Translating nociceptor sensitivity: the role of axonal protein synthesis in nociceptor physiology. Eur. J. Neurosci., 29, 2253–2263.
Guan, X., Zhu, X. and Tao, Y. X. (2009) Peripheral nerve injury up-regulates expression of interactor protein for cytohesin exchange factor 1 (IPCEF1) mRNA in rat dorsal root ganglion. Naunyn Schmiedebergs Arch. Pharm-acol., 380, 459–463.
Singh, O. V., Yaster, M., Xu, J. T., Guan, Y., Guan, X., Dharmarajan, A. M., Raja, S. N., Zeitlin, P. L. and Tao, Y. X. (2009) Proteome of synaptosome-associated proteins in spinal cord dorsal horn after peripheral nerve injury. Proteomics 9, 1241–1253.
Yang, L., Zhang, F. X., Huang, F., Lu, Y. J., Li, G. D., Bao, L., Xiao, H. S. and Zhang, X. (2004) Peripheral nerve injury induces trans-synaptic modification of channels, receptors and signal pathways in rat dorsal spinal cord. Eur. J. Neurosci., 19, 871–883.
Coyle, D. E. (2007) Spinal cord transcriptional profile analysis reveals protein trafficking and RNA processing as prominent processes regulated by tactile allodynia. Neuroscience 144, 144–156.
Acknowledgments
This work was supported by National Institutes of Health Grants NS 058886 and NS 072206 and the Blaustein Pain Research Fund (Y.-X. T). The authors thank Claire Levine, MS, for her editorial assistance.
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Singh, O.V., Tao, YX. (2012). Two-Dimensional Gel Electrophoresis: Discovering Neuropathic Pain-Associated Synaptic Biomarkers in Spinal Cord Dorsal Horn. In: Luo, Z. (eds) Pain Research. Methods in Molecular Biology, vol 851. Humana Press. https://doi.org/10.1007/978-1-61779-561-9_3
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DOI: https://doi.org/10.1007/978-1-61779-561-9_3
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Online ISBN: 978-1-61779-561-9
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