Polgár Erika, Hughes David I, Arham Ahmad Z, Todd Andrew J
Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, United Kingdom.
J Neurosci. 2005 Jul 13;25(28):6658-66. doi: 10.1523/JNEUROSCI.1490-05.2005.
It has been proposed that death of inhibitory interneurons in the dorsal horn contributes to the neuropathic pain that follows partial nerve injury. In this study, we have used two approaches to test whether there is neuronal death in the dorsal horn in the spared nerve injury (SNI) model. We performed a stereological analysis of the packing density of neurons in laminas I-III 4 weeks after operation and found no reduction on the ipsilateral side compared with that seen on the contralateral side or in sham-operated or naive rats. In addition, we used two markers of apoptosis, terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling (TUNEL) staining and immunocytochemical detection of cleaved (activated) caspase-3. Neither of these methods demonstrated apoptotic neurons in the dorsal spinal cord 1 week after operation. Although TUNEL-positive cells were present throughout the gray and white matter at this stage, they were virtually all labeled with antibody against ionized calcium-binding adapter molecule 1, a marker for microglia. All animals that underwent SNI showed clear signs of tactile allodynia affecting the ipsilateral hindpaw. These results suggest that a significant loss of neurons from the dorsal horn is not necessary for the development of tactile allodynia in the SNI model.
有人提出,脊髓背角抑制性中间神经元的死亡会导致部分神经损伤后出现的神经性疼痛。在本研究中,我们采用了两种方法来测试在 spared nerve injury(SNI)模型中脊髓背角是否存在神经元死亡。我们在术后4周对I-III层神经元的堆积密度进行了体视学分析,发现与对侧、假手术组或未处理的大鼠相比,同侧没有减少。此外,我们使用了两种凋亡标记物,即末端脱氧核苷酸转移酶介导的生物素化UTP缺口末端标记(TUNEL)染色和裂解(活化)的caspase-3的免疫细胞化学检测。这两种方法均未在术后1周的脊髓背角中显示出凋亡神经元。尽管在此阶段TUNEL阳性细胞存在于整个灰质和白质中,但它们几乎都被抗离子钙结合衔接分子1(一种小胶质细胞标记物)的抗体标记。所有接受SNI手术的动物均表现出明显的触觉异常性疼痛迹象,影响同侧后爪。这些结果表明,在SNI模型中,脊髓背角神经元的显著丧失对于触觉异常性疼痛的发展并非必要条件。