YAP蛋白在神经肌肉接头中作用机制的研究  

Study on the mechanism of YAP protein in neuromuscular junction

作　　者：杨滨瑞 赵琳琳[1] 汪煜楠 初婷婷[1] 郑永慧[1] 张惊宇[1] YANG Bin rui;ZHAO Lin-lin;WANG Yu-nan(Department of Neurology,the Fourth Affiliated Hospital of Harbin Medical Univerity.Harbrin 150001,China)

机构地区：[1]哈尔滨医科大学附属第四医院神经内科,黑龙江哈尔滨150001

出　　处：《中国实验诊断学》2023年第5期603-607,共5页Chinese Journal of Laboratory Diagnosis

基　　金：国家自然科学基金面上项目(项目编号:62272138);横向课题(项目编号:HX2020-12)。

摘　　要：目的探讨YAP蛋白对神经肌肉接头的影响及其机制。方法以质粒为模板构建YAP敲低的shRNA,用Western Blot方法检测shYAP的敲低效率,并在此基础上将shYAP包进AAV8并感染C2C12肌管,通过免疫荧光染色检测乙酰胆碱受体的长度和荧光强度。将agrin加入至分化成熟的C2C12肌管中,通过免疫荧光染色检测加入agrin后细胞核中YAP的表达情况和乙酰胆碱受体聚集情况。采用Western Blot方法检验样本在agrin处理后YAP磷酸化的合成水平和YAP结合β-环联蛋白的生成体量。结果YAP shRNA构建成功;肌管形成后敲低YAP能抑制乙酰胆碱受体聚集;agrin能促进C2C12肌管中的YAP进入细胞核;agrin能减弱YAP蛋白S127位的磷酸化。结论YAP作为agrin下游信号分子调节神经肌肉接头形成。Objective To investigate the effect of YAP protein on neuromuscular junction and its mechanism.Methods The plasmid was used as template to construct shRNA with YAP knockdown.Western Blot method was used to detect the knockdown efficiency of shYAP.Based on this,shYAP was encapsulated into AAV8 and infected with C2C12 myoduct.Agrin was added into the differentiated and mature C2C12 myotube,and the expression of YAP and aggregation of acetylcholine receptor in the nucleus of agrin was detected by immunofluorescence staining.Western Blot was used to determine the synthesis level of YAP phosphorylation and the production volume of YAP-bindingβ-cyclin after agrin treatment.Results YAP shRNA was constructed successfully.Knocking down YAP after myoduct formation can inhibit the aggregation of acetylcholine receptor.Agrin can promote YAP in C2C12 myotube to enter the nucleus.Agrin attenuates the phosphorylation of YAP at S127.Conclusion YAP as a downstream signal molecule of AGRin regulates neuromuscular junction formation.

关 键 词：神经肌肉接头 Yes相关蛋白 聚集蛋白 

分 类 号：R741.05[医药卫生—神经病学与精神病学]