CRMP2在大鼠海马神经元中的表达分布及对神经元突起生长的影响

目的探讨脑衰反应调节蛋白2(CRMP2)在大鼠海马神经元中的表达分布及对神经元突起生长的影响。方法CRMP2及其模拟磷酸化质粒CRMP2-S522D、CRMP2-T555D,转入体外培养的SD大鼠海马神经元,转染72 h后,采用免疫印迹法和免疫荧光染色分析CRMP2、CRMP2-S522D、CRMP2-T555D在神经元中的分布差异以及对神经元突起分枝和突起长度的影响。结果 CRMP2及CRMP2-T555D在神经元胞体和突起均有分布,CRMP2-S522D则主要分布于胞体。CRMP2-T555D对神经元的突起长度和分枝数均没有明显影响;CRMP2-S522D对突起数目没有明显影响,但可以显著降低突起长度。结论 CRMP2的不同磷酸化形式在大鼠海马神经元中存在分布差异,并对神经元的突起长度产生影响。

Distribution of different phosphorylated forms of CRMP2 in hippocampal neurons and their effect on neurites growth of hippocampal neurons in rats

Objective To study the distribution of Ser-522 and Thr-555 phosphorylated forms of collapsin response mediator protein (CRMP) 2 in the hippocampal neurons and their effects on neurite of those neurons. Methods The plasmids of CRMP2-S522D and CRMP2-T555D were constructed and transferred into hippocampal neurons which were derived from wildtype Sprague-Dawley rat and cultured in vitro. The neurons were fixed and stained 72 hours after the transfection and then observed by laser scanning confocal microscope. The analysis of the distribution of CRMP2, CRMP2-S522D and CRMP2-T555D in the neurons and their effects on the neurons neurite elongation and branching were analyzed. Results CRMP2 and CRMP2-T555D were distributed in the neuronal bodies and neurites and CRMP2-S522D was mainly distributed in the neuronal bodies. CRMP2-T555D had no significant effect on the neurite elongation and branching. CRMP2-S522D had no significant effect on the number of the neurons neurites, but could significantly reduce their neurite length. Conclusion The different phosphorylated forms of CRMP2 were distributed in different cellular structures of the hippocampal neurons in the rat and CRMP2-S522D may influence the neurons neurite elongation.