Wu Xiaofang, Yoo Soonmoon, Wrathall Jean R
Department of Neuroscience, Georgetown University Medical Center, Washington, District of Columbia 20057, USA.
J Neurochem. 2005 May;93(4):943-52. doi: 10.1111/j.1471-4159.2005.03078.x.
Rat spinal cord contusion injury models the histopathology associated with much clinical spinal cord injury (SCI). Studies on altered gene expression after SCI in these models may identify therapeutic targets for reducing secondary injury after the initial trauma and/or enhancing recovery processes. However, complex spatial and temporal alterations after injury could complicate interpretation of changes in gene expression. To test this hypothesis, we selected six genes and studied their temporal and spatial patterns of expression at 1 h, 1, 3 and 7 days after a standardized spinal cord contusion produced by a weight drop device (10 g x 25 mm at T8). Real-time RT-PCR using TaqMan probes was employed to quantify mRNA for proteolipid protein, glyceraldehyde-3-phosphate dehydrogenase, glial fibrillary acidic protein, nestin, and the GluR2 and NR1 subunits of glutamate receptors. We found widely different temporal and spatial patterns of altered gene expression after SCI, including instances of opposing up- and down-regulation at different locations in tissue immediately adjacent to the injury site. We conclude that greater use of the reliable and extremely sensitive technique of quantitative real-time PCR for regional tissue analysis is important for understanding the altered gene expression that occurs after CNS trauma.
大鼠脊髓挫伤模型可模拟许多临床脊髓损伤（SCI）相关的组织病理学变化。对这些模型中脊髓损伤后基因表达改变的研究，可能会确定减少初始创伤后继发性损伤和/或促进恢复过程的治疗靶点。然而，损伤后复杂的时空变化可能会使基因表达变化的解释变得复杂。为了验证这一假设，我们选择了六个基因，并研究了在通过重物坠落装置（T8处10 g×25 mm）造成标准化脊髓挫伤后1小时、1天、3天和7天它们的时空表达模式。使用TaqMan探针的实时逆转录聚合酶链反应（RT-PCR）用于定量检测蛋白脂质蛋白、甘油醛-3-磷酸脱氢酶、胶质纤维酸性蛋白、巢蛋白以及谷氨酸受体的GluR2和NR1亚基的信使核糖核酸（mRNA）。我们发现脊髓损伤后基因表达改变的时空模式差异很大，包括在紧邻损伤部位的组织中不同位置出现上调和下调相反的情况。我们得出结论，更多地使用可靠且极其灵敏的定量实时PCR技术进行区域组织分析，对于理解中枢神经系统创伤后发生的基因表达改变非常重要。
