用于神经突导向研究的螺旋神经元培养体系初步观察  

Neurites of spiral ganglion neurons suiting for neurite guidance investigation in vitro

作　　者：李树峰[1] 王正敏[1] 李雯[2] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻喉科,上海200031 [2]复旦大学附属眼耳鼻喉科医院实验中心,上海200031

出　　处：《中国眼耳鼻喉科杂志》2011年第2期82-85,139,140,共6页Chinese Journal of Ophthalmology and Otorhinolaryngology

基　　金：上海市卫生局青年科研项目资助(2008Y100)

摘　　要：目的探寻适用于螺旋神经元神经突导向研究的螺旋神经元培养体系。方法取新生SD大鼠的螺旋神经节,分别进行分离培养和组织块培养,比较神经突的生长模式。组织块培养时,采用不同底物(包括多聚赖氨酸、层粘连蛋白和I型胶原蛋白等)铺片,观察对螺旋神经元组织块贴壁和神经突生长的影响。用微管相关蛋白2(MAP2)抗体和微管聚合蛋白1(Tau-1)抗体鉴别神经突的极性。结果组织块培养体系中,部分神经突长出细胞毯,在无干扰的环境中生长。以多聚赖氨酸加层粘连蛋白为底物铺片时,组织块贴壁率高,神经突数量多、生长较快,有利于获取更多适于神经突导向研究的神经突。培养体系中的神经突可被Tau-1抗体标记,而不能被MAP2抗体标记。结论在多聚赖氨酸加层粘连蛋白包被的可移动玻片上,进行螺旋神经元组织块培养获取自由神经突的方法适合螺旋神经元神经突的导向研究。Objective To investigate a method of ceil culture suiting for neurite guidance research of spiral gan- glion neurons. Methods The authors explored the extension and development of spiral ganglion neurites from neonatal Sprague-Dawley rats by either explant culture or disassociated culture. In addition, the impact of different substratums （including poly-D-lysine, laminin and collagen I） on neurite extension was studied. To differentiate dendrites and axons, immunofluorescence for microtubule associated protein 2 （MAP2） and Tau-1 was observed. Results There were a few neurites that grew out of the cell blanket to a clean area in the explant culture of spiral ganglion neurons on movable slides. The direction of neurite extension appeared random because there was no interference from other cells, which was a characteristic suiting for the investigation of neurite guidance. Explants on poly-D-lysine plus laminin had relatively higher adhesion ratio, more and longer ＂free＂ neuritis than explants on other substratum. The neurites in the spiral ganglion explants could be labeled by anti-Tau-1 but not anti-MAP2. Conclusions Incubating free neurites on poly-D-lysine plus laminin-coated movable slides was suitable for neurite guidance research of spiral ganglion neurons in vitro. The polarity of neurites in explant culture system was immature.

关 键 词：螺旋神经元 神经突导向 神经突 组织块培养 多聚赖氨酸 层粘连蛋白 

分 类 号：R764[医药卫生—耳鼻咽喉科]