Liu Zheng, Huang Wei, Zhu Man-Min, Xu Zhong-Xiang, Xu Zu-Cai, Yu Chang-Yin, Huang Hao
Department of Neurology Affiliated Hospital of Zunyi Medical University Zunyi China.
Ibrain. 2023 Aug 15;9(4):369-380. doi: 10.1002/ibra.12127. eCollection 2023 Winter.
The neural network hypothesis is one of the important pathogenesis of drug-resistant epilepsy. Axons guide molecules through synaptic remodeling and brain tissue remodeling, which may result in the formation of abnormal neural networks. Therefore, axon guidance plays a crucial role in disease progression. However, although Robo1 is one of the important components of axon guidance, the role of Robo1 in epilepsy remains unclear. In this study, we aimed to explore the mechanism of Robo1 in epilepsy. Male adult C57BL/6 mice were intraperitoneally injected with pentylenetetrazol to establish an epilepsy model. Lentivirus (LV) was given via intracranial injection 2 weeks before pentylenetetrazol injection. Different expressions of Robo1 between the control group, LV-mediated Robo1 short hairpin RNA group, empty vector control LV group, and normal saline group were analyzed using Western blot, immunofluorescence staining, Golgi staining, and video monitoring. Robo1 was increased in the hippocampus in the pentylenetetrazol-induced epilepsy mouse model; lentiviral Robo1 knockdown prolonged the latency of seizure and reduced the seizure grade in mice and resulted in a decrease in dendritic spine density, while the number of mature dendritic spines was maintained. We speculate that Robo1 has been implicated in the development and progression of epilepsy through its effects on dendritic spine morphology and density. Epileptic mice with Robo1 knockdown virus intervention had lower seizure grade and longer latency. Follow-up findings suggest that Robo1 may modulate seizures by affecting dendritic spine density and morphology. Downregulation of Robo1 may negatively regulate epileptogenesis by decreasing the density of dendritic spines and maintaining a greater number of mature dendritic spines.
神经网络假说为耐药性癫痫的重要发病机制之一。轴突通过突触重塑和脑组织重塑来引导分子，这可能导致异常神经网络的形成。因此，轴突导向在疾病进展中起着关键作用。然而，尽管Robo1是轴突导向的重要组成部分之一，但其在癫痫中的作用仍不清楚。在本研究中，我们旨在探讨Robo1在癫痫中的作用机制。成年雄性C57BL/6小鼠腹腔注射戊四氮以建立癫痫模型。在注射戊四氮前2周通过颅内注射给予慢病毒（LV）。使用蛋白质免疫印迹法、免疫荧光染色、高尔基染色和视频监测分析对照组、LV介导的Robo1短发夹RNA组、空载体对照LV组和生理盐水组之间Robo1的不同表达。在戊四氮诱导的癫痫小鼠模型中，海马体中的Robo1增加；慢病毒介导的Robo1基因敲低可延长小鼠癫痫发作的潜伏期并降低癫痫发作等级，导致树突棘密度降低，而成熟树突棘的数量保持不变。我们推测，Robo1通过影响树突棘的形态和密度参与了癫痫的发生和发展。接受Robo1基因敲低病毒干预的癫痫小鼠癫痫发作等级较低，潜伏期较长。后续研究结果表明，Robo1可能通过影响树突棘密度和形态来调节癫痫发作。Robo1的下调可能通过降低树突棘密度并维持更多数量的成熟树突棘来对癫痫发生起负向调节作用。
