Singh Om V, Tao Yuan-Xiang
Division of Biological and Health Science, University of Pittsburgh-Bradford, Pittsburgh, PA, USA.
Methods Mol Biol. 2012;851:47-63. doi: 10.1007/978-1-61779-561-9_3.
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.
神经损伤诱导的神经性疼痛是全球范围内的一个主要公共卫生问题。目前针对神经性疼痛的治疗效果有限，因为引发和维持神经性疼痛的机制尚未完全明确。然而，蛋白质组学的最新进展可能使我们能够揭示在神经性疼痛条件下发生的复杂生物学机制。在此，我们介绍一种二维凝胶电泳（2-DE）与质谱（MS）相结合的方法，以鉴定单侧第五脊髓神经损伤后脊髓背角突触体相关蛋白的表达变化。在二维凝胶电泳中，一组等电点范围为4至7的高度丰富的突触蛋白通过大小分级分离（25至100 kDa）并进行比较。然后，通过质谱鉴定和验证差异表达的蛋白质，并探索它们在生理和病理过程中的潜在作用。因此，蛋白质组学分析可以提供神经性疼痛条件下神经系统细胞、组织和器官中突触蛋白的表达谱及其翻译后修饰情况。
