Blomgren K, McRae A, Bona E, Saido T C, Karlsson J O, Hagberg H
Institute of Anatomy and Cell Biology, Göteborg University, Sweden.
Brain Res. 1995 Jul 3;684(2):136-42. doi: 10.1016/0006-8993(95)00398-a.
Neonatal rats were subjected to transient cerebral hypoxic-ischemia (unilateral occlusion of the common carotid artery + 7.70% O2 for 100 min) and allowed to recover for 3 h, 24 h, 2 days or 14 days. Consecutive tissue sections were stained with antibodies against alpha-fodrin, the 150 kDa breakdown product of alpha-fodrin (FBDP, marker of calpain proteolysis) or microtubule associated protein 2 (MAP 2, marker of dendrosomatic neuronal injury). Cortical tissue pieces were subjected to Western blotting using the antibody against the FBDP. Areas with brain injury displayed a distinct loss of MAP 2 which clearly delineated the infarct. FBDP accumulated in injured and borderline regions ipsilaterally and a less conspicuous, transient increase in FBDP also occurred in the contralateral hemisphere, especially in the white matter. A reciprocal staining pattern could be seen in the cerebral cortex, i.e. loss of MAP 2 and accumulation of FBDP, most pronounced 14 days after the insult. Fodrin and MAP 2 are known calpain substrates, and degradation of these proteins preceded neuronal degeneration, indicating that these proteases may be involved in the early events triggering the cascades leading to neuronal death.
新生大鼠经历短暂性脑缺氧缺血（单侧颈总动脉闭塞 + 7.70%氧气，持续100分钟），并使其恢复3小时、24小时、2天或14天。连续的组织切片用抗α-血影蛋白、α-血影蛋白150 kDa降解产物（FBDP，钙蛋白酶解标志物）或微管相关蛋白2（MAP 2，树突体神经元损伤标志物）的抗体进行染色。使用抗FBDP抗体对皮质组织块进行蛋白质印迹分析。脑损伤区域显示MAP 2明显缺失，清晰勾勒出梗死灶。FBDP在同侧损伤区域和边缘区域积聚，对侧半球尤其是白质中也出现不太明显的FBDP短暂增加。在大脑皮质中可观察到相反的染色模式，即MAP 2缺失和FBDP积聚，在损伤后14天最为明显。血影蛋白和MAP 2是已知的钙蛋白酶底物，这些蛋白质的降解先于神经元变性，表明这些蛋白酶可能参与触发导致神经元死亡的级联反应的早期事件。
