中枢神经系统与周围神经系统发育和损伤过程中微管动态性的差异比较  

Comparative study on microtubule dynamics in central nervous system and peripheral nervous system during development and injury

作　　者：柏为娟 马超[2] 巫荣华 刘炎[2] 刘梅[2] BO Weijuan;MA Chao;WU Ronghua;LIU Yan;LIU Mei(Blood Testing Laboratory of Yancheng City Blood Center,Jiangsu Province,Yancheng 224005;Key Laboratory of Neuroregeneration of Jiangsu and Ministry of Education,Co-innovation Center of Neuroregeneration,Nantong University)

机构地区：[1]江苏省盐城市中心血站血液检测中心,盐城224005 [2]南通大学教育部/江苏省神经再生重点实验室,神经再生协同创新中心

出　　处：《南通大学学报（医学版）》2022年第6期497-501,共5页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(32070725)。

摘　　要：目的:通过比较微管蛋白的酪氨酸化、去酪氨酸化水平,分析中枢神经和周围神经系统神经元发育和损伤后微管动态变化。方法:分离培养Sprague-Dawley(SD)大鼠孕18 d(E18)胎鼠的背根神经节神经元和海马神经元,采用免疫细胞荧光化学方法检测不同修饰微管蛋白的表达和定位,对培养的神经元进行划伤,采用活细胞拍摄对神经元胞体及轴突生长进行分析比较。提取SD大鼠生后发育不同时间点的坐骨神经和脊髓组织,同时建立SD大鼠的脊髓挫伤和坐骨神经夹伤模型,于术后不同时间提取蛋白,通过Western Blot方法检测不同修饰(酪氨酸化、去酪氨酸化)的微管蛋白表达情况。结果:免疫细胞化学结果显示,与神经元种植1 d相比,在3 d时,背根神经节神经元,酪氨酸化、去酪氨酸化微管蛋白均维持较高水平,而海马神经元中酪氨酸化微管蛋白表达较低;活细胞摄像观察显示,背根神经节神经元和海马神经元在体外发育和损伤再生过程中轴突的平均生长速率差异无统计学意义,但背根神经节神经元胞体运动速率显著高于海马神经元胞体。中枢神经和周围神经发育过程中,坐骨神经组织维持较高的酪氨酸化微管蛋白水平,而在脊髓组织中酪氨酸化微管蛋白的表达逐步下降;在神经损伤后,酪氨酸化微管蛋白在坐骨神经组织中表达显著高于脊髓酪氨酸化微管蛋白。结论:与中枢神经相比,周围神经在发育和损伤后酪氨酸化修饰微管蛋白水平均高于中枢神经,提示周围神经具有更强的微管动态性。Objective:To analyze the dynamic changes of microtubules during neuronal development and injury in the central and peripheral nervous systems by comparing the levels of tyrosinylation,detyrosinylation of tubulins.Methods:The dorsal root ganglion neurons and hippocampal neurons were isolated and cultured from embryonic day 18(E18)fetuses of pregnant Sprague-Dawleg(SD)rats.Immunocytofluorescence was used to detect the expression and localization of different modified tubulins.Scratching the live cell imaging was used to analyze and compare neuronal cell bodies motility and axonal growth.The sciatic nerve and spinal cord tissues of SD rats were collected at different time points after birth,and the models of spinal cord contusion and sciatic nerve crush were established in SD rats.Protein were extracted at different times after surgery.The expression of tubulins with different modifications(tyrosinylation,detyrosinylation)was detected by Western Blot.Results:Immunocytochemical results showed that compared with the 1 d cultured neurons,the levels of tyrosinated,detyrosinated tubulins were maintained at higher levels in dorsal root ganglion neurons,while tyrosinated tubulins were maintained at lower levels in hippocampal neurons at 3 d.Live cell imaging shows that the average axonal growth rate of dorsal root ganglion neurons and hippocampal neurons during in vitro development and injury had no significance.However,there were significant changes in cell body motility.The cell body movement rate of dorsal root ganglion neurons was significantly higher than that of hip-pocampal neurons.During the development of central and peripheral nervous system,sciatic nerve tissue maintained high levels of tyrosinated tubulins,while the expression of tyrosinated tubulins in spinal cord tissue decreased;after injury,in sci-atic nerve tissue,expression of tyrosinated tubulins significantly increased.Conclusion:The level of tyrosinated tubulins in the peripheral nervous system is higher than that in the central nervous system after de

关 键 词：酪氨酸化微管蛋白 去酪氨酸化微管蛋白 海马神经元 背根神经节神经元 脊髓 坐骨神经 神经损伤 大鼠 

分 类 号：R651[医药卫生—外科学]