Spinal SNAP-25 regulates membrane trafficking of GluA1-containing AMPA receptors in spinal injury-induced neuropathic pain in rats
Spinal SNAP-25 regulates membrane trafficking of GluA1-containing AMPA receptors in spinal injury-induced neuropathic pain in rats
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脊髓 SNAP-25 在脊髓损伤引起的大鼠神经性疼痛中调节含 GluA1 AMPA 受体的膜运输
DOI：
10.1016/j.neulet.2019.134616
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发表时间：
2020-01-10
影响因子：
2.5
通讯作者：
Zhang, Linlin
中科院分区：
文献类型：
--
作者：
Liu, Peng;Song, Chengcheng;Zhang, Linlin
简介：突触体相关蛋白25 kDa（SNAP-25）是可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体复合物的一个成员，对膜融合和神经递质的释放至关重要。α-氨基-3-羟基-5-甲基-4-异恶唑丙酸（AMPA）受体与病理性疼痛有关。本研究旨在探讨SNAP-25是否以及如何调节AMPA受体在神经病理性疼痛中的作用。方法：雄性Sprague道利大鼠进行L-4脊神经结扎（SNL）或假手术。在评估机械异常性疼痛和热敏感性后，收获L4-5脊髓的同侧部分。采用Western blot和实时定量聚合酶链反应分析SNAP-25的表达水平。用Western印迹分析评估SNAP-25磷酸化和AMPA受体膜运输水平。SNAP-25表达或磷酸化抑制证实了SNAP-25和AMPA膜运输之间的关联。结果：SNL程序诱导并维持机械异常性疼痛和热痛觉过敏。SNL增加SNAP-25的表达和磷酸化以及AMPA受体在脊髓中的膜运输。SNAP-25表达或磷酸化抑制减轻SNL后神经病理性疼痛并下调AMPA受体的膜运输。GluA 1抑制AMPA受体可减轻SNL.Conclusions后的机械异常性疼痛和热痛觉过敏：SNL诱导的神经病理性疼痛是通过SNAP-25丝氨酸187位磷酸化引起的SNAP-25依赖的AMPA受体膜转运的上调。因此，SNAP-25表达或磷酸化的抑制可用作神经性疼痛的治疗。然而，含GluA 1的AMPA受体膜运输的SNAP-25磷酸化介导的神经病理性疼痛的机制值得进一步探讨。
Introduction: Synaptosomal associated proteins of 25 kDa (SNAP-25), as a member of stable soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex, is critical for membrane fusion and required for the release of neurotransmitters. The alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor is implicated in pathologic pain. This study aimed to investigate whether and how SNAP-25 regulated AMPA receptors in neuropathic pain.Methods: Male Sprague Dawley rats underwent L-4 spinal nerve ligation (SNL) or the sham procedure. After assessing mechanical allodynia and thermal sensitivity, the ipsilateral portion of the L4-5 spinal cord was harvested. The expression level of SNAP-25 was analyzed by Western blot analysis and real-time quantitative polymerase chain reaction. SNAP-25 phosphorylation and AMPA receptor membrane trafficking levels were evaluated with Western blot analysis. An association between SNAP-25 and AMPA membrane trafficking was confirmed by SNAP-25 expression or phosphorylation inhibition.Results: The SNL procedure induced and maintained mechanical allodynia and thermal hyperalgesia. SNL increased the expression and phosphorylation of SNAP-25 and the membrane trafficking of AMPA receptors in the spinal cord. SNAP-25 expression or phosphorylation inhibition alleviated neuropathic pain and downregulated membrane trafficking of AMPA receptors after SNL. GluA1-containing AMPA receptor inhibition relieved mechanical allodynia and thermal hyperalgesia after SNL.Conclusions: The upregulation of SNAP-25-dependent membrane trafficking of AMPA receptors via SNAP-25 phosphorylation at Ser187 contributed to SNL-induced neuropathic pain. Thus, the inhibition of SNAP-25 expression or phosphorylation might serve as a treatment for neuropathic pain. However, the mechanism of GluA1-containing AMPA receptor membrane trafficking mediated by SNAP-25 phosphorylation in neuropathic pain deserves further exploration.