Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on differentiating mouse N2a neuroblastoma cells
Effects of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine on differentiating mouse N2a neuroblastoma cells
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DOI：
10.1046/j.1471-4159.2000.0750133.x
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发表时间：
2000-07-01
影响因子：
4.7
通讯作者：
Hargreaves, AJ
中科院分区：
文献类型：
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作者：
De Girolamo, LA;Billett, EE;Hargreaves, AJ
研究了神经毒素 1-甲基-4-苯基-1,2,3,6-四氢吡啶 (MPTP) 在小鼠 N2a 神经母细胞瘤细胞中的作用，通过在 24 小时内撤除血清并添加二丁酰环 AMP 诱导其分化。在分化过程中添加MPTP（10μM）引起细胞形态的变化，其特征是在不存在细胞死亡的情况下抑制轴突生长。蛋白质印迹的生化表征表明，MPTP 对肌动蛋白、α-微管蛋白或总重链神经丝 (NF-H) 的水平没有显着影响。然而，当用磷酸化状态特异性抗体 RMd09 和 Ta51 探测印迹时，MPTP 处理后 NF-H 磷酸化似乎增加。此外，间接免疫荧光分析显示磷酸化 NF-H 在细胞核周中积累，表明 NF-H 分布的改变与观察到的 MPTP 对细胞形态的影响有关。这些变化可能代表简单分化神经元细胞模型系统中 MPTP 神经毒性的有用体外标记。
The effect of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) was investigated in mouse N2a neuroblastoma cells, induced to differentiate by serum withdrawal and addition of dibutyryl cyclic AMP, over a 24-h period. Addition of MPTP (10 mu M) during differentiation caused a change in cell morphology characterised by an inhibition of axon outgrowth, in the absence of cell death. Biochemical characterisation by western blotting revealed that MPTP had no significant effects on the levels of actin, alpha-tubulin, or total heavy-chain neurofilament (NF-H). However, NF-H phosphorylation appeared to increase following MPTP treatment when blots were probed with the phosphorylation state-specific antibodies RMd09 and Ta51. In addition, indirect immunofluorescence analysis revealed an accumulation of phosphorylated NF-H in the cell perikaryon, suggesting that altered NF-H distribution was associated with the observed effects of MPTP on cell morphology. These changes may represent a useful in vitro marker of MPTP neurotoxicity within a simple differentiating neuronal cell model system.