Physiology Department, Ribeirão Preto School of Medicine, University of São Paulo, Ribeirão Preto, São Paulo, Brazil.
Brain Res. 2011 Feb 16;1374:43-55. doi: 10.1016/j.brainres.2010.12.012. Epub 2010 Dec 11.
The aims of this study were to characterize the spatial distribution of neurodegeneration after status epilepticus (SE) induced by either systemic (S) or intrahippocampal (H) injection of pilocarpine (PILO), two models of temporal lobe epilepsy (TLE), using FluoroJade (FJ) histochemistry, and to evaluate the kinetics of FJ staining in the H-PILO model. Therefore, we measured the severity of behavioral seizures during both types of SE and also evaluated the FJ staining pattern at 12, 24, and 168 h (7days) after the H-PILO insult. We found that the amount of FJ-positive (FJ+) area was greater in SE induced by S-PILO as compared to SE induced by H-PILO. After SE induced by H-PILO, we found more FJ+ cells in the hilus of the dentate gyrus (DG) at 12 h, in CA3 at 24 h, and in CA1 at 168 h. We found also no correlation between seizure severity and the number of FJ+ cells in the hippocampus. Co-localization studies of FJ+ cells with either neuronal-specific nuclear protein (NeuN) or glial fibrillary acidic protein (GFAP) labeling 24 h after H-PILO demonstrated spatially selective neurodegeneration. Double labeling with FJ and parvalbumin (PV) showed both FJ+/PV+ and FJ+/PV- cells in hippocampus and entorhinal cortex, among other areas. The current data indicate that FJ+ areas are differentially distributed in the two TLE models and that these areas are greater in the S-PILO than in the H-PILO model. There is also a selective kinetics of FJ+ cells in the hippocampus after SE induced by H-PILO, with no association with the severity of seizures, probably as a consequence of the extra-hippocampal damage. These data point to SE induced by H-PILO as a low-mortality model of TLE, with regional spatial and temporal patterns of FJ staining.
这项研究的目的是使用 FluoroJade (FJ) 组织化学方法，描述由全身（S）或海马内（H）注射匹罗卡品（PILO）引起的癫痫持续状态（SE）后神经退行性变的空间分布，这两种模型都是颞叶癫痫（TLE），并评估 H-PILO 模型中 FJ 染色的动力学。因此，我们测量了两种 SE 期间行为性癫痫发作的严重程度，还评估了 H-PILO 损伤后 12、24 和 168 小时（7 天）的 FJ 染色模式。我们发现，与 H-PILO 诱导的 SE 相比，S-PILO 诱导的 SE 中 FJ 阳性（FJ+）区域的数量更大。在 H-PILO 诱导的 SE 后，我们发现 12 小时时 DG 齿状回（DG）的棘旁细胞更多，24 小时时 CA3 区更多，168 小时时 CA1 区更多。我们还发现，海马中 FJ+细胞的数量与癫痫发作的严重程度之间没有相关性。H-PILO 后 24 小时 FJ+细胞与神经元特异性核蛋白（NeuN）或胶质纤维酸性蛋白（GFAP）标记的共定位研究表明存在空间选择性神经退行性变。FJ 与 parvalbumin（PV）的双重标记显示海马和内嗅皮质等区域存在 FJ+/PV+和 FJ+/PV-细胞。目前的数据表明，FJ+区域在两种 TLE 模型中的分布不同，在 S-PILO 模型中比在 H-PILO 模型中更大。H-PILO 诱导的 SE 后，FJ+细胞也存在选择性动力学，与癫痫发作的严重程度无关，可能是由于海马外损伤所致。这些数据表明，H-PILO 诱导的 SE 是一种低死亡率的 TLE 模型，具有 FJ 染色的区域空间和时间模式。
