Wang Jian, Zhang Fan, Luo Zhong, Zhang Haiqing, Yu Changyin, Xu Zucai
Department of Neurology, Affiliated Hospital of Zunyi Medical University, Zunyi, Guizhou 563000, China.
Department of Neurology, Affiliated Aerospace Hospital of Zunyi Medical University, Zunyi, Guizhou 563000, China.
Genes Dis. 2024 Mar 19;11(6):101266. doi: 10.1016/j.gendis.2024.101266. eCollection 2024 Nov.
Abnormal mitochondrial dynamics can lead to seizures, and improved mitochondrial dynamics can alleviate seizures. Vacuolar protein sorting 13D (VPS13D) is closely associated with regulating mitochondrial homeostasis and autophagy. However, further investigation is required to determine whether VPS13D affects seizures by influencing mitochondrial dynamics and autophagy. We aimed to investigate the influence of VPS13D on behavior in a rat model of acute epileptic seizures. Hence, we established an acute epileptic seizure rat model and employed the CRISPR/CAS9 technology to construct a lentivirus to silence the gene. Furthermore, we used the HT22 mouse hippocampal neuron cell line to establish a stable strain with suppressed expression of . Then, we performed quantitative proteomic and bioinformatics analyses to confirm the mechanism by which VPS13D influences mitochondrial dynamics and autophagy, both and using the experimental acute epileptic seizure model. We found that knockdown of resulted in reduced seizure latency and increased seizure frequency in the experimental rats. Immunofluorescence staining and western blot analysis revealed a significant increase in mitochondrial dynamin-related protein 1 expression following knockdown. Moreover, we observed a significant reduction in LC3II protein expression levels and the LC3II/LC3I ratio (indicators for autophagy) accompanied by a significant increase in P62 expression (an autophagy adaptor protein). The proteomic analysis confirmed the up-regulation of P62 protein expression. Therefore, we propose that VPS13D plays a role in modulating seizures by influencing mitochondrial dynamics and autophagy.
异常的线粒体动力学可导致癫痫发作，而改善线粒体动力学可减轻癫痫发作。液泡蛋白分选13D（VPS13D）与调节线粒体稳态和自噬密切相关。然而，需要进一步研究以确定VPS13D是否通过影响线粒体动力学和自噬来影响癫痫发作。我们旨在研究VPS13D对急性癫痫发作大鼠模型行为的影响。因此，我们建立了急性癫痫发作大鼠模型，并采用CRISPR/CAS9技术构建慢病毒以沉默该基因。此外，我们使用HT22小鼠海马神经元细胞系建立了一个稳定株，其表达受到抑制。然后，我们进行了定量蛋白质组学和生物信息学分析，以证实VPS13D影响线粒体动力学和自噬的机制，在实验性急性癫痫发作模型中均进行了研究。我们发现，在实验大鼠中敲低该基因导致癫痫发作潜伏期缩短和癫痫发作频率增加。免疫荧光染色和蛋白质印迹分析显示，敲低该基因后，线粒体动力相关蛋白1的表达显著增加。此外，我们观察到LC3II蛋白表达水平和LC3II/LC3I比值（自噬指标）显著降低，同时P62表达（一种自噬衔接蛋白）显著增加。蛋白质组学分析证实了P62蛋白表达上调。因此，我们提出VPS13D通过影响线粒体动力学和自噬在调节癫痫发作中发挥作用。
