Spreading depression and selective neuronal death

抑郁症的蔓延和选择性神经元死亡

• 批准号: 07671685
• 负责人: IIJIMA Takehiko
• 金额: $ 1.66万
• 依托单位: Kyorin University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07671685/
• 关键词: spreading depression; brain ischemia; glutamate; MAP2; SNAP-25; hippocampus; brain resuscitation; microdialysis; 選択的神経細胞死; マイクロダイアライシス; 海馬CA1; 脳血流量

Our research results are divided into three parts ; 1) MAP2 depletion in the hippocampus following spreading depression 2) SNAP-25 expression after repeated induction of spreading depression 3) glutamate release during spreading depression1) MAP2 depletion in the hippocampus following spreading depressionTraumatic brain injury induces neuronal cell loss in area CA3 of the hippocampus. However, it has not yet been established why trauma of the cortex induces neuroral damage in a remote area. Spreading depression (SD) may be one potential mechanism for this pathophysiology. The present study evaluated whether SD on the cortex evokes a pathological change in the hippocampus. Forty-two Fisher rats were assigned to four groups : Group I : sham operation (n=7), Group II : right carotid occlusion (UO) for 7days. (n=7), GroupIII : repeated induction of SD by KCl application on dura for 7 days (n=7), GroupIII'for 3hours (n=7), Group IV : SD induction and UO for seven days (n=14) GroupIV' for 3h … More ours (n=7) In five out of 7 animals in GroupsIII' and IV', cerebral blood flow was monitored using laser Doppler flowmetry for 3h during the passage of SD.The brains were pocessed for immunohistochemical analysis of microtubule-associated protein 2 (MAP2). Reactive hyperemia induced by SD was not significantly suppressed by right carotid occlusion (194(]SY+-[))25% and 181(]SY+-[))42% UO in Groups III and IV,respectively). In six out of seven animals in 7 days model of Group IV,and three animals in 7 days model of Group III,MAP2 depletion in the CA3 area of the hippocampus (partly including CA2) was observed, although no change in the hippocampus was observed in other groups. In conclution, SD in combination with UO yielded reproducible lesions in CA3. Neuronal injury in the hippocampus after brain trauma may be attributable to SD in combination with the blood flow restriction.2) SNAP-25 expression after repeated induction of spreading depressionRepeated induction of cortical spreading depression (SD) elicits neuronal cell loss in area CA3 of the hippocampus, when the blood flow is restricted by unilateral carotid occlusion. This model may be able to extrapolated to remote neuronal injury due to traumatic brain injury. In this study, we evaluated whether cortical SD augmented hippocampal glutamate release by immunohistochemical analysis of SNAP-25, an exocytosis-related protein. Fifteen rats were assigned to three groups of five : I ; control ; II ; SD induction and unilateral carotid artery occlusion (UO) for 3h ; III ; SD and UO for seven days. The brains were processed for immunohistochemical analysis of SNAP-25 and MAP2. Two of the five animals in Group II showed MAP2 depletion in the CA3 area of the hippocampus (including partly of CA2), was observed, whereas hippocampal changes were observed in Group I.Fairly strong SNAP-25 immunoreactivity present, in the area corresponding to the MAP2 depleted region in Group II.Four of the five Group III animals showed MAP2 depletion in CA-3 of the hippocampus, but, no distinct SNAP-25 immunostaining was observed, not even in the MAP2 depletion area. In conclusion, glutamate release in CA-3 may be augmented by cortically induced SD on the cortex for three hours. Remote neuronal injury in the hippocampus resulting from cortically induced SD may be modulated by glutamate release.3) glutamate release during spreading depressionSpreading depression (SD) seems to play a key role in extending neuronal damage in the penumbra. SD alone does not elicit neuronal damage, so supplemental factors which might turn SD into a harmful stimulus were examined. Eleven male Fischer rats were anesthetized and mechanically ventilated in a stereotaxic frame. A laser Doppler flowmeter, an electroenzymatic electrode for measuring glutamate and a calomel electrode for measuring DC potential were placed through a cranial window positioned 3 mm away from a second window where KCl soaked cotton was placed to initiate SD.The left carotid artery was ligated to suppress reactive hyperemia of SD.The electroenzymatic electrode continuously monitored extracellular glutamate which was oxidized by L-glutamate oxidase perfused through the electrode. Glutamate levels increased immediately on initiation of SD from 34(]SY+-[)24 mM/L to 48(]SY+-[)47 mM/L (p<0.05) before carotid occlusion, from 41(]SY+-[)25 mM/L to 53(]SY+-[)41 mM/L (p<0.05) after ligation of the carotid artery, returned to the control value when the DC potential recovered. There was no positive correlation between the duration of SD and suppression of reactive hyperemia, however, a positive correlation (p<0.05) was found between the duration of SD and the glutamate concentration. In conclusion, high extracellular glutamate levels accompany prolonged SD.Prolonged SD in ischemic penumbra may be caused by high extracelluar glutamate. Less

我们的研究成果分为三个部分； 1) 扩散性抑郁症后海马 MAP2 耗竭 2) 重复诱导扩散性抑郁症后 SNAP-25 表达 3) 扩散性抑郁症期间谷氨酸释放 1) 扩散性抑郁症后海马 MAP2 耗竭 创伤性脑损伤导致海马 CA3 区神经元细胞损失。然而，目前尚不清楚为什么皮质创伤会引起偏远区域的神经元损伤。抑郁扩散（SD）可能是这种病理生理学的一种潜在机制。本研究评估了皮质上的 SD 是否会引起海马体的病理变化。 42只Fisher大鼠被分为四组：第一组：假手术（n=7），第二组：右颈动脉闭塞（UO）7天。 (n=7)，第 III 组：通过在硬脑膜上应用 KCl 重复诱导 SD 7 天 (n=7)，第 III 组’持续 3 小时 (n = 7)，第 IV 组：SD 诱导和 UO 7 天 (n = 14) 第 IV 组’持续 3 小时……更多我们的 (n = 7) 在第 III 组和第 IV 组的 7 只动物中，有 5 只使用激光多普勒监测脑血流 在SD通过期间进行流量测定3小时。对大脑进行微管相关蛋白2（MAP2）的免疫组织化学分析。 SD 引起的反应性充血并没有被右颈动脉闭塞显着抑制（第 III 组和第 IV 组分别为 194(]SY+-[))25% 和 181(]SY+-[))42% UO）。在第IV组的7天模型中的7只动物中的6只和第III组的7天模型中的3只动物中，观察到海马CA3区域（部分包括CA2）中MAP2的缺失，但在其他组中没有观察到海马的变化。总之，SD 与 UO 组合在 CA3 中产生了可重复的损伤。脑外伤后海马神经元损伤可能归因于SD与血流限制相结合。2）重复诱导扩散性抑制后SNAP-25的表达当血流受到单侧颈动脉闭塞限制时，重复诱导皮质扩散性抑制（SD）会引起海马CA3区神经元细胞损失。该模型可能能够推断出由于创伤性脑损伤而导致的远程神经元损伤。在本研究中，我们通过对 SNAP-25（一种胞吐作用相关蛋白）进行免疫组织化学分析，评估了皮质 SD 是否会增加海马谷氨酸的释放。十五只老鼠被分配到三组，每组五只：I；I；I。控制 ;二、 SD诱导并单侧颈动脉闭塞（UO）3小时；三、 SD 和 UO 持续 7 天。对大脑进行处理以进行 SNAP-25 和 MAP2 的免疫组织化学分析。第二组的五只动物中的两只显示出海马 CA3 区域（包括部分 CA2）中的 MAP2 缺失，而第一组中观察到海马变化。在对应于第二组中 MAP2 缺失区域的区域中存在相当强的 SNAP-25 免疫反应性。第三组中的五只动物中的四只显示出海马 CA-3 中的 MAP2 缺失， 但是，没有观察到明显的 SNAP-25 免疫染色，甚至在 MAP2 缺失区域也没有观察到。总之，皮质诱导的 SD 持续三小时可能会增强 CA-3 中谷氨酸的释放。皮质诱导的 SD 导致的海马远端神经元损伤可能受到谷氨酸释放的调节。3) 扩散性抑郁期间谷氨酸的释放 扩散性抑郁 (SD) 似乎在延长半暗带神经元损伤中发挥着关键作用。单独的 SD 不会引起神经元损伤，因此检查了可能将 SD 变成有害刺激的补充因素。将 11 只雄性 Fischer 大鼠麻醉并在立体定位框架中进行机械通气。将激光多普勒流量计、用于测量谷氨酸的电酶电极和用于测量直流电势的甘汞电极通过距离第二个窗口3mm的颅窗放置，其中放置KCl浸泡的棉花以启动SD。结扎左颈动脉以抑制SD的反应性充血。电酶电极连续监测细胞外谷氨酸 它被通过电极灌注的L-谷氨酸氧化酶氧化。 SD 开始后，谷氨酸水平立即从颈动脉闭塞前的 34(]SY+-[)24 mM/L 增加至 48(]SY+-[)47 mM/L (p<0.05)，颈动脉结扎后从 41(]SY+-[)25 mM/L 增加至 53(]SY+-[)41 mM/L (p<0.05)，返回到对照直流电势时的值 恢复了。 SD持续时间与反应性充血抑制之间不存在正相关，但SD持续时间与谷氨酸浓度之间存在正相关（p<0.05）。总之，高细胞外谷氨酸水平伴随着SD的延长。缺血半暗带中SD的延长可能是由高细胞外谷氨酸引起的。较少的

项目成果

Iijama T.,Shimase C.,Sawa H.,Sankawa H: "Repeated induction of spreading depression and exocytosis-related protein in the hippocampus" submitted to Advances in Neurotrauma Research. 9. (1997)

Iijama T.、Shimase C.、Sawa H.、Sankawa H：“在海马中重复诱导扩散抑郁和胞吐作用相关蛋白”，提交给《神经创伤研究进展》。

Iijima T.,Shimase C.,Sawa H.,Sankawa H.: "Repeated induction of spreading depression and exocytosis-related protein in the hippocampus" submitted to Advances in Neurotrauma Research. 9. (1997)

Iijima T.、Shimase C.、Sawa H.、Sankawa H.：“在海马中重复诱导扩散抑郁和胞吐作用相关蛋白”，提交给《神经创伤研究进展》。

Iijima T., Shimase C., Sankawa H: "Relationship between glutamate release, blood flow and spreading depression : real-time monitoring using an electroenzymatic dialysis electrode." Journal of Cerebral Blood Flow and Metabolism. (submitted to).

Iijima T.、Shimase C.、Sankawa H：“谷氨酸释放、血流和扩散性抑郁之间的关系：使用电酶透析电极进行实时监测。”

Iijima T., Sawa H., Shiokawa Y., Saito I., Ishii H., Nakamura Z., Sankawa H: "Thromboxane synthetase inhibitor ameliorates delayd neuronal death in the CA1 subfield of the hippocampus after transient global ischemia in gerbils." J Neurosug Anesthesiology.

Iijima T.、Sawa H.、Shiokawa Y.、Saito I.、Ishii H.、Nakamura Z.、Sankawa H：“血栓烷合成酶抑制剂可改善沙鼠短暂性整体缺血后海马 CA1 亚区的延迟性神经元死亡。”

Iijima T.,Iijima C.,Shimase C.,Sankawa H.: "Glutamate release following global ischemia differs between reting and cerebral cortex in the rat." J Cerebral Blood Flow and Metab. Suppl 1. (1997)

Iijima T.、Iijima C.、Shimase C.、Sankawa H.：“大鼠全身缺血后的谷氨酸释放在大鼠的视网膜和大脑皮层之间存在差异。”