β-微管蛋白-Ⅲ在脊髓神经干细胞向神经元定向分化早期的动态变化研究

Dynamic changes of β-tubulin-Ⅲ in the early directed differentiation from the spinal cord neural stem cells to neurons

目的研究脊髓神经干细胞定向分化为神经元过程中细胞骨架蛋白——β-微管蛋白-Ⅲ(β-tubulin-Ⅲ)的表达及意义。方法采用细胞培养技术、免疫细胞化学(SABC)技术鉴定培养细胞,免疫荧光技术观察β-微管蛋白-Ⅲ在脊髓神经干细胞定向分化为神经元过程中的分布情况;采用Western blotting检测β-微管蛋白-Ⅲ在神经干细胞分化过程中的变化。结果分化第1天,β-微管蛋白-Ⅲ表达较弱;随着细胞的发育,分化第4天时β-微管蛋白-Ⅲ表达增强,其阳性反应物分布于核周,并伸出突起;β-微管蛋白-Ⅲ的含量也随着细胞的生长而增多;然而分化第7天时,β-微管蛋白-Ⅲ表达减少。结论在脊髓神经干细胞定向分化为神经元过程中,细胞骨架蛋白——β-微管蛋白-Ⅲ影响和促进细胞的分化。

Objective To study about the expression and significance of β-Tubulin-m in directed differentiation of neural cells from spinal neural stem cells. Methods Cell culture technology, immunocytochemistry （SABC） technique were used to identify the cultured cells. The distributions of β- Tubulin-Ⅲ by immunofluorescence were observed in the process of the spinal cord neural stem cell differentiation of neurons. In addition, we detected β-Tubulin- Ⅲ by Western blotting. Results There was weak expression of β-Tubulin- Ⅲ in the first day. During the growth of neuron, the expression of β-Tubulin- Ⅲ was increasing and distributed around the cell nucleus and into the process in the 4th day. The content of β- Tubulin-Ⅲ was also increasing with the growth of neurons. However, the expression of β-Tubulin-Ⅲ in the 7th day was lower than the 4th day. Conclusions During directed differentiation of spinal derived-neural stem cells to neurons, β-Tubulin- Ⅲ effect and promote the differentiation of neuron.