Stretch-induced injury in organotypic hippocampal slice cultures reproduces in vivo post-traumatic neurodegeneration:: role of glutamate receptors and voltage-dependent calcium channels
Stretch-induced injury in organotypic hippocampal slice cultures reproduces in vivo post-traumatic neurodegeneration:: role of glutamate receptors and voltage-dependent calcium channels
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DOI：
10.1111/j.1471-4159.2006.04379.x
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发表时间：
2007-04-01
影响因子：
4.7
通讯作者：
Sundstrom, Lars E.
中科院分区：
文献类型：
--
作者：
Cater, Heather L.;Gitterman, Daniel;Sundstrom, Lars E.
在体内，导致脑组织变形的初始机械事件与延迟性神经退行性变之间的关系很复杂，因为涉及多种因素。我们使用了一种基于在可变形硅膜上培养的大鼠海马切片的简化脑替代物来研究拉伸诱导的创伤性脑损伤。通过拉伸培养底物诱导创伤性损伤，并在4天后对生物学反应进行表征。在受损的培养物中广泛观察到与人类创伤性损伤一致的形态异常。严重损伤后突触功能显著降低。N -甲基 - D -天冬氨酸（NMDA）受体拮抗剂MK - 801减轻了神经元损伤，防止了微管相关蛋白2免疫反应性的丧失，并减轻了突触功能的降低。相比之下，NMDA受体拮抗剂3 - [（R） - 2 - 羧基哌嗪 - 4 - 基] - 丙基 - 1 - 膦酸（CPP）和GYKI53655在中度损伤模式下具有神经保护作用，但在重度损伤模式下则无。硝苯地平，一种L型电压依赖性钙通道拮抗剂，仅在中度损伤后具有保护作用，而ω - 芋螺毒素在重度损伤后减轻了损伤。这些结果表明，拉伸损伤后的损伤机制很复杂，并且因损伤的严重程度而异。总之，器官型海马切片培养物对拉伸损伤的药理学、形态学和电生理学反应与在体内观察到的相似。我们的模型为理解创伤后延迟性细胞死亡的机制提供了一种替代动物实验的方法，并且可以在进入体内模型之前用作一种高内涵筛选来发现神经保护化合物。
The relationship between an initial mechanical event causing brain tissue deformation and delayed neurodegeneration in vivo is complex because of the multiplicity of factors involved. We have used a simplified brain surrogate based on rat hippocampal slices grown on deformable silicone membranes to study stretch-induced traumatic brain injury. Traumatic injury was induced by stretching the culture substrate, and the biological response characterized after 4 days. Morphological abnormalities consistent with traumatic injury in humans were widely observed in injured cultures. Synaptic function was significantly reduced after a severe injury. The N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 attenuated neuronal damage, prevented loss of microtubule-associated protein 2 immunoreactivity and attenuated reduction of synaptic function. In contrast, the NMDA receptor antagonists 3-[(R)-2-carboxypiperazin-4-yl]-propyl-1-phosphonic acid (CPP) and GYKI53655, were neuroprotective in a moderate but not a severe injury paradigm. Nifedipine, an L-type voltage-dependent calcium channel antagonist was protective only after a moderate injury, whereas omega-conotoxin attenuated damage following severe injury. These results indicate that the mechanism of damage following stretch injury is complex and varies depending on the severity of the insult. In conclusion, the pharmacological, morphological and electrophysiological responses of organotypic hippocampal slice cultures to stretch injury were similar to those observed in vivo. Our model provides an alternative to animal testing for understanding the mechanisms of post-traumatic delayed cell death and could be used as a high-content screen to discover neuroprotective compounds before advancing to in vivo models.