P2X7 receptor immunoreactive profile confined to resting and activated microglia in the epileptic brain
P2X7 receptor immunoreactive profile confined to resting and activated microglia in the epileptic brain
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DOI：
10.1016/j.brainres.2006.03.040
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发表时间：
2006-05
期刊：
影响因子：
2.9
通讯作者：
P. Rappold;E. Lynd-Balta;E. Lynd-Balta;S. Joseph
中科院分区：
文献类型：
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作者：
P. Rappold;E. Lynd-Balta;E. Lynd-Balta;S. Joseph
本研究的目的是鉴定kainate诱导的癫痫发作和非癫痫发作大鼠脑CNS细胞成分对特异性p2x7受体抗血清的免疫反应。p2x7免疫细胞化学（ICC）分析显示，随着癫痫发作时间的推移，小的免疫反应细胞的过程显示出明显的形态变化。这些形态学变化使人想起中枢神经系统损伤时的反应性胶质细胞。为了确定这种非神经元细胞成分的身份，我们采用了双ICC技术，使用顺序抗体孵育，并将切片与对比色原反应。该系列检测的特异性胶质细胞标志物包括Iba1（小胶质细胞）、COX-1（小胶质细胞）和GFAP（星形胶质细胞）。当p2x7免疫染色切片用小胶质细胞特异性抗血清（Iba1, COX-1）进行双重免疫染色时，该研究结果显示了明显的共定位。相反，当切片用p2x7和GFAP（一种星形细胞标记物）进行双重免疫染色时，没有明显的共定位。在后一种实验中，双ICC显示了两种不同的细胞群，具有对比色，显示出不同的GFAP免疫阳性细胞群和不同的p2x7免疫阳性细胞群。我们得出结论，本研究中使用的p2x7抗血清对大鼠的小胶质细胞具有特异性，并且在盐酸盐诱导的癫痫发作后，小胶质细胞形态存在进行性变化的时间轴。此外，癫痫诱导后小胶质细胞的形态变化可以用p2x7抗血清或小胶质细胞特异性抗血清识别，这表明中枢神经系统癫痫活动区域存在神经炎症环境。
The purpose of this study was to identify the CNS cellular constituent immunoreactive for specific P2X7receptor antiserum in the kainate-induced seizure and non-seizure rat brain. Analysis of P2X7immunocytochemistry (ICC) revealed small immunoreactive cells with processes showing distinct morphological changes as seizures progressed in time. These morphological changes were reminiscent of reactive glia during CNS injury. In order to determine the identity of this non-neuronal cellular constituent, we employed dual ICC techniques using sequential antibody incubations and reacted the sections with contrasting chromagens. Specific glial markers tested in the series included Iba1 (microglia), COX-1 (microglia), and GFAP (astroglia). Results of this study revealed distinct colocalization when sections immunostained for P2X7were dual immunostained with antisera specific for microglia (Iba1, COX-1). In contrast, no colocalization was evident when sections were dual immunostained with P2X7and GFAP, an astrocytic marker. In the latter experiment, dual ICC revealed two distinct cell populations with contrasting color demonstrating a population of distinct GFAP immunopositive cells and a population of distinct P2X7immunopositive cells. We conclude that P2X7antiserum used in this study is specific for and identifies microglia in rat and that there exists a timeline of progressive changes in microglia morphology that can be demonstrated following kainate-induced seizures. In addition, the morphological changes in microglia following seizure induction that can be identified with P2X7antisera or with antisera specific for microglia suggest a neuroinflammatory milieu in areas of CNS seizure activity.