Hampton D W, Rhodes K E, Zhao C, Franklin R J M, Fawcett J W
Cambridge Centre for Brain Repair, E. D. Adrian Building, Forvie Site, Robinson Way, Cambridge CB2 2PY, UK.
Neuroscience. 2004;127(4):813-20. doi: 10.1016/j.neuroscience.2004.05.028.
The cortical stab injury has been widely used for biochemical analysis of molecular changes following CNS injury. However, the cellular responses to this injury have not been accurately quantified. In order to provide a baseline for biochemical studies and future experiments on the manipulation of the CNS injury response we have undertaken a quantitative analysis of this injury. The proliferative and reactive responses of oligodendrocyte precursor cells, astrocytes and microglia were measured, using antibodies to NG2, glial fibrillary acidic protein (GFAP) and the cd11-b clone OX-42 to characterise these cell types at 2, 4, 7 and 14 days post-injury. Oligodendrocyte precursors and microglia proliferated rapidly during the first week, mostly within 0.3 mm of the lesion. Of the dividing cells over 60% were oligodendrocyte precursor cells with microglia making up the balance of the dividing cells. Minimal numbers of astrocytes divided in response to the lesion. Large cells with one or two short processes that were both NG2 and OX-42 positive were identified very close to the lesion at 2 and 4 days post-lesion but not thereafter. They are likely to be blood-derived cells that express NG2 or have ingested it. NG2 immunohistochemistry and platelet-derived growth factor alpha receptor (PDGFalpha-R) in situ hybridisation on neighbouring sections was performed. In the lesioned area only 12% of NG2 positive (+ive) cells were PDGFalpha-R +ive (a ratio of 1:8 for PDGFalpha-R +ive cells: NG2 +ive cells) compared with 33% in the unlesioned cortex and an almost 100% overlap in the spinal cord.
皮质刺伤已被广泛用于中枢神经系统(CNS)损伤后分子变化的生化分析。然而,对这种损伤的细胞反应尚未进行准确量化。为了为生化研究以及未来关于操纵CNS损伤反应的实验提供基线,我们对这种损伤进行了定量分析。使用针对NG2、胶质纤维酸性蛋白(GFAP)和cd11-b克隆OX-42的抗体,在损伤后2、4、7和14天测量少突胶质前体细胞、星形胶质细胞和小胶质细胞的增殖和反应性反应,以表征这些细胞类型。少突胶质前体细胞和小胶质细胞在第一周迅速增殖,主要在损伤部位0.3毫米范围内。在分裂细胞中,超过60%是少突胶质前体细胞,小胶质细胞占分裂细胞的其余部分。对损伤作出反应而分裂的星形胶质细胞数量极少。在损伤后2天和4天,在非常靠近损伤部位处发现了具有一个或两个短突起且NG2和OX-42均呈阳性的大细胞,但此后未再发现。它们可能是表达NG2或摄取了NG2的血液来源细胞。在相邻切片上进行了NG2免疫组织化学和血小板衍生生长因子α受体(PDGFalpha-R)原位杂交。在损伤区域,只有12%的NG2阳性(+ive)细胞是PDGFalpha-R +ive(PDGFalpha-R +ive细胞与NG2 +ive细胞的比例为1:8),而在未损伤的皮质中这一比例为33%,在脊髓中几乎有100%的重叠。