Wang Xueying, Zhou Liqian, Yang Lin, Huang Shaoping, Wang Yuying, Li Dan
Department of Pediatrics, The Second Affliated Hospital of Xi'an Jiaotong University, Xi'an, China.
Department of radiology, ShaanXi Provincial People's Hospital, Xi'an, China.
Mol Neurobiol. 2025 May 10. doi: 10.1007/s12035-025-04971-w.
The objective of this study was to elucidate the molecular mechanisms by which cAMP-regulated transcription coactivator1 (CRTC1) regulates autophagy and GluA2 expression in patients with epilepsy.
We initially established a magnesium-free epilepsy cell model and recorded cellular discharges using the whole-cell patch clamp technique. Next, we experimentally activated autophagy and identified effective methods for silencing the CRTC1 gene using RNA interference technology. Furthermore, we developed an animal models of status epilepticus and employed immunofluorescence and Western Blot to elucidate CRTC1's role in regulating autophagy-related genes and GluA2 expression in epilepsy.
We observed mouse hippocampal neurons under magnesium-free extracellular conditions. Treatment with an autophagy activator decreased GluA2 expression; however, CRTC1 was not dephosphorylated. CRTC1 siRNA suppressed LC3 and PSD95 expression, whereas CRTC1 siRNA intervention restored GluA2 expression.
CRTC1 indirectly influences the expression of synaptic-related proteins and GluA2 by directly modulating autophagy during the pathological process of epilepsy. The findings of this study reveal novel molecular targets for the treatment of epilepsy.
本研究的目的是阐明环磷酸腺苷调节的转录共激活因子1（CRTC1）在癫痫患者中调节自噬和谷氨酸受体2（GluA2）表达的分子机制。
我们首先建立了无镁癫痫细胞模型，并使用全细胞膜片钳技术记录细胞放电。接下来，我们通过实验激活自噬，并使用RNA干扰技术确定沉默CRTC1基因的有效方法。此外，我们建立了癫痫持续状态动物模型，并采用免疫荧光和蛋白质印迹法阐明CRTC1在癫痫中调节自噬相关基因和GluA2表达的作用。
我们在无镁细胞外条件下观察小鼠海马神经元。用自噬激活剂处理可降低GluA2表达；然而，CRTC1未发生去磷酸化。CRTC1小干扰RNA抑制微管相关蛋白1轻链3（LC3）和突触后致密物95（PSD95）表达，而CRTC1小干扰RNA干预可恢复GluA2表达。
在癫痫的病理过程中，CRTC1通过直接调节自噬间接影响突触相关蛋白和GluA2的表达。本研究结果揭示了治疗癫痫的新分子靶点。
