p150glued缺失对神经元中Rab蛋白表达水平的影响

Effects of p150glued deficiency on Rab proteins expression in cultured cortical neurons

目的探讨家族性运动神经元病致病基因DCTN1编码的p150glued蛋白缺失对于原代培养的皮质神经元细胞活性及Rab蛋白表达的影响。方法以野生型(WT)和p150glued缺失(p150glued KO)新生小鼠为细胞来源,体外培养皮质神经元7d和14d后,分别利用MTT法和Western blot法检测皮质神经元存活率以及Rab蛋白家族成分表达水平。结果与WT小鼠皮质神经元相比,p150glued KO小鼠皮质神经元在体外培养7d和14d时,细胞活力无明显差异,14d时Rab4、Rab5、Rab9、Rab11、Rab24蛋白表达水平未发生显著变化,而Rab7蛋白表达水平显著降低(P<0.001)。结论 p150glued缺失对原代皮质神经元存活以及调控囊泡运输的Rab蛋白家族中Rab4、Rab5、Rab9、Rab11、Rab24的表达水平无明显影响,但特异性减弱晚期内体标志物Rab7蛋白的表达,可阻碍胞内溶酶体降解通路。

Objective To investigate the effects of motor neuron disease(MND)related DCTN1 gene encoded p150glued protein deficiency on cell viability and expression of Rab proteins in cultured mouse cortical neurons.Method The cortical neurons from wild type(WT)and p150glued knock out(p150glued KO)newborn mice were cultured in vitro for 7 to 14 days.The cell viability and the expressions of Rab proteins were detected with MTT assay and Western blot,respectively.Results Compared to WT mice,cortical neurons from p150glued KO mice showed comparable cell viability on 7 and 14 days in vitro.On 14 days in vitro,cortical neurons from p150glued KO mice showed decreased level of Rab7 only(P<0.001).Conclusion p150glued deficiency specifically reduces the expression of vesicular transport related protein Rab7 in cultured cortical neurons.