Valproic Acid Influences MTNR1A Intracellular Trafficking and Signaling in a β-Arrestin 2-Dependent Manner
Valproic Acid Influences MTNR1A Intracellular Trafficking and Signaling in a β-Arrestin 2-Dependent Manner
复制标题
丙戊酸以 β-Arrestin 2 依赖性方式影响 MTNR1A 细胞内运输和信号传导
DOI：
10.1007/s12035-014-9085-y
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发表时间：
2016-03-01
影响因子：
5.1
通讯作者：
Han, Feng
中科院分区：
文献类型：
--
作者：
Hong, Ling-juan;Jiang, Quan;Han, Feng
丙戊酸暴露与自闭症谱系障碍的风险增加有关。迄今为止，褪黑激素受体亚型1 （MTNR1A）内化在丙戊酸暴露后的机制细节仍然难以捉摸。通过在HEK-293和neuro2a细胞中表达表位标记受体（MTNR1A- egfp），我们记录了褪黑激素处理后MTNR1A细胞内运输的动态变化。使用延时共聚焦显微镜，我们在活细胞中发现，在褪黑激素存在的情况下，丙戊酸干扰了MTNR1A的内化动力学。这种减弱作用与PKA （Thr197）和ERK （Thr202/Tyr204）磷酸化的降低有关。VPA处理没有改变褪黑素或不褪黑素细胞的全细胞电流。此外，荧光共振能量转移成像数据表明，丙戊酸降低了褪黑激素引发的yfp标记的β-阻滞蛋白2和cfp标记的MTNR1A之间的关联。总之，我们认为丙戊酸以β-阻滞蛋白2依赖的方式影响MTNR1A细胞内运输和信号传导。
Valproate exposure is associated with increased risks of autism spectrum disorder. To date, the mechanistic details of disturbance of melatonin receptor subtype 1 (MTNR1A) internalization upon valproate exposure remain elusive. By expressing epitope-tagged receptors (MTNR1A-EGFP) in HEK-293 and Neuro-2a cells, we recorded the dynamic changes of MTNR1A intracellular trafficking after melatonin treatment. Using time-lapse confocal microscopy, we showed in living cells that valproic acid interfered with the internalization kinetics of MTNR1A in the presence of melatonin. This attenuating effect was associated with a decrease in the phosphorylation of PKA (Thr197) and ERK (Thr202/Tyr204). VPA treatment did not alter the whole-cell currents of cells with or without melatonin. Furthermore, fluorescence resonance energy transfer imaging data demonstrated that valproic acid reduced the melatonin-initiated association between YFP-labeled β-arrestin 2 and CFP-labeled MTNR1A. Together, we suggest that valproic acid influences MTNR1A intracellular trafficking and signaling in a β-arrestin 2-dependent manner.