摘要
目的 :探讨微管稳定剂埃博霉素D(Epo D)对孤独症谱系障碍BTBR小鼠皮质神经元兴奋性突触结构的影响及机制。方法 :体外培养BTBR小鼠原代大脑皮质神经元,免疫荧光法检测所培养神经元的纯度;培养至相对成熟的BTBR小鼠皮质神经元随机分为实验组(Epo D组)、溶剂对照组(0.1%DMSO)干预24 h,再冷处理90min后行微管免疫荧光染色,观察微管的形态变化;免疫印迹检测微管稳定的标志蛋白(acetyl-tubulin、α-tubulin)、微管相关蛋白(MAP2、STOP)的表达,及兴奋性突触结构前、后膜标志蛋白(VGLUT1、PSD95)、兴奋性谷氨酸受体相关蛋白(GluN2B、mGluR5)的表达水平。结果 :与对照组相比,10 nmol/L的Epo D可增加BTBR小鼠皮质神经元微管的冷稳定性及微管稳定的标志蛋白、微管相关蛋白的表达;可以增加BTBR小鼠皮质神经元兴奋性突触结构、兴奋性谷氨酸受体相关蛋白的表达,差异均有统计学意义。结论 :微管稳定剂Epo D能改善体外培养的BTBR小鼠皮质神经元兴奋性突触结构,其机制可能与增加微管稳定性有关。
Objective : To investigate the effect and mechanism of microtubule stabilizer Epothilone D on the excitatory synaptic structure of cortical neurons in BTBR mouse with autism spectrum disorder. Methods : The primary cerebral cortical neurons of BTBR mouse were isolated and cultured, and the purity of cultured neurons was detected by immunofluorescence technology. Cortical neurons of relatively mature were randomly divided into an experimental group(epothilone D group) and a vehicle control group(0.1% v/v DMSO) for 24 h intervention.After cold treatment for 90 min, the morphological changes of microtubules were observed by microtubule immunofluorescence staining. Western blotting was used to detect the expression of acetyl-tubulin, α-tubulin, MAP2, STOP, VGLUT1, PSD95, GluN2B and mGluR5 proteins. Results :Compared with the control group, the cold stability of microtubules in cortical neuron of BTBR mouse was increased with 10 nmol/L Epothilone D.The expressions of excitatory synapse-related and excitatory glutamate receptor-related proteins of cortical neurons in BTBR mouse were increased. Conclusion : The microtubule stabilizer epothilone D can improve the excitatory synaptic structure of cortical neurons in BTBR mouse in vitro, which may be related to the increase of microtubule stability.
作者
陈永红
杨桦
刘永峰
雷强
蔚洪恩
Chen Yonghong;Yang Hua;Liu Yongfeng;Lei Qiang;Wei Hongen(Department of Neurology,The Fifth Clinical Medical College,Shanxi Medical University,Taiyuan 030012,China;Basic Medical College,Shanxi Medical University,Taiyuan 030012,China)
出处
《解剖学杂志》
CAS
2022年第3期222-227,共6页
Chinese Journal of Anatomy
基金
国家自然科学基金(81671364)
山西省卫生健康委员会科研项目(2020TD25,2020XM32,2020RC09)。
