Reactive astrocytosis from excitotoxic injury in hippocampal organ culture parallels that seen in vivo
Reactive astrocytosis from excitotoxic injury in hippocampal organ culture parallels that seen in vivo
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DOI：
10.1097/00004647-199701000-00005
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发表时间：
1997-01-01
影响因子：
6.3
通讯作者：
Kraig, RP
中科院分区：
文献类型：
--
作者：
Kunkler, PE;Kraig, RP
反应性星形胶质细胞不仅影响脑损伤的严重程度，还影响大脑通过学习重塑自身的能力。负责星形胶质细胞增生的机制仍然未知，但可能是最好的研究在体外，其中改进的访问和可视化允许应用现代分子和细胞技术。我们已经开始探索是否可以在海马器官型培养物（HOTC）中研究神经胶质增生，其中保留了潜在的细胞间相互作用，并且仍然实现了体外制备的优点。HOTC暴露于N-甲基-D-天冬氨酸（NMDA）后，星形胶质细胞免疫组织化学[免疫染色（IS）]标记物胶质细胞酸性蛋白（GFAP）和波形蛋白的光密度（OD）值发生剂量依赖性变化。HOTC暴露于NMDA（10 μ M）引起CA（1）海马区和齿状回的选择性死亡。它还显著增加了这些区域的GFAP IS和波形蛋白IS OD值。在CA（3）中也观察到GFAP IS和波形蛋白IS OD值增加，海马区域未显示细胞死亡。光学显微镜检查显示GFAP和波形蛋白IS细胞肥大，为反应性星形胶质细胞的特征。细胞增殖，增殖细胞核抗原IS评估，也显着增加，在所有这三个海马区。相比之下，HOTC暴露于非损伤性水平的NMDA（1 μ M）引起的GFAP IS和波形蛋白IS OD值只有轻微的变化，但在所有HOTC地区的细胞增殖显着减少。这些结果表明HOTC兴奋毒性损伤引起的反应性星形胶质细胞增多与全脑缺血后体内观察到的变化相似。此外，由于HOTC内的静息星形胶质细胞也类似于它们在体内的对应物，因此HOTC可用于检查这些细胞在类似于完整脑的组织内转化为反应性物质的机制。
Reactive astrocytes influence not only the severity of brain injury, but also the capacity of brain to reshape itself with learning. Mechanisms responsible for astrogliosis remain unknown but might be best studied in vitro, where improved access and visualization permit application of modern molecular and cellular techniques. We have begun to explore whether gliosis might be studied in hippocampal organotypic cultures (HOTCs), where potential cell-to-cell interactions are preserved and the advantages of an in vitro preparation are still realized. Following HOTC exposure to N-methyl-D-aspartate (NMDA), dose-dependent changes occurred in the optical density (OD) values for the astrocytic immunohistochemical [immunostaining (IS)] markers glial fibrillary acidic protein (GFAP) and vimentin. Exposure of HOTCs to NMDA (10 mu M) caused selective death in the CA(1) hippocampal re ion and the dentate gyrus. It also significantly increased GFAP IS and vimentin IS OD values in these regions. Increased GFAP IS and vimentin IS OD values were also seen in CA(3), a hippocampal region that displayed no cell death, Light microscopic examination revealed hypertrophied GFAP and vimentin IS cells, characteristic of reactive astrocytes. Cellular proliferation, as assessed by proliferating cell nuclear antigen IS, was also significantly increased in all three of these hippocampal regions. In contrast, exposure of HOTCs to a noninjurious level of NMDA (1 mu M) caused only minor changes in GFAP IS and vimentin IS OD values but a significantly reduced cellular proliferation in all HOTC regions. These results show that reactive astrocytosis from excitotoxic injury of HOTC parallels changes seen in vivo after global ischemia. Furthermore, since resting astroglia within HOTCs an also similar to their counterparts in vivo, HOTCs may be used to examine mechanisms by which these cells are transformed into reactive species within tissue that resembles intact brain.