首页|背根神经节在Nav1.7相关先天性疼痛不敏感中的变化及作用

背根神经节在Nav1.7相关先天性疼痛不敏感中的变化及作用

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目的 探究背根神经节(dorsal root ganglion,DRG)神经元在电压门控钠通道1.7功能缺失(voltage-gated sodium channel 1.7 loss-of-function,Nav1.7 LOF)大鼠中的变化及其参与先天性疼痛不敏感的机制.方法 利用免疫荧光法、Western blot法、可变剪切分析验证Nav1.7 LOF大鼠DRG神经元与脊髓Nav1.7的表达缺失情况.在DRG、脊髓组织中使用外周蛋白(peripherin)、降钙素基因相关肽(calcitonin gene-related peptide,CGRP)、异凝集素 B4(isolectin B4,IB4)、神经元核抗原(NeuN)抗体,免疫荧光染色法观察Nav1.7 LOF大鼠DRG神经元亚群及其中枢末梢投射与野生型大鼠的差异.利用全转录组测序(bulk RNA-Seq)与实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RT-qPCR)检测Nav1.7 LOF与野生型大鼠DRG组织基因表达差异.55℃热刺激Nav1.7 LOF大鼠后爪,利用原癌基因(c-Fos)与磷酸化细胞外信号调节激酶(phospho-extracellular signal-regulated kinase,p-ERK)抗体染色检查DRG神经元及脊髓背角神经元的激活情况.结果 Nav1.7 LOF大鼠DRG与脊髓Nav1.7缺失.与野生型大鼠比较,Nav1.7 LOF大鼠DRG组织IB4+非肽能神经元比例下降,IB4+神经元中枢末梢在脊髓背角的投射显著缺失.RNA-Seq和RT-qPCR检测结果显示,Nav1.7 LOF大鼠IB4+非肽能神经元亚群标志物Mas相关G蛋白耦联受体(Mas-related G protein-coupled receptors,MRGPR)基因表达下调.Nav1.7 LOF大鼠后爪接受热刺激后DRG神经元c-Fos、p-ERK染色与野生型对照无差异,但脊髓背角神经元无c-Fos、p-ERK染色,提示该神经元未被激活.结论 在Nav1.7 LOF大鼠中,IB4+非肽能神经元的比例及其中枢末梢投射显著下降,伤害性刺激可激活DRG神经元,但不能激活脊髓背角神经元,即疼痛信号在DRG神经元的中枢末梢与脊髓背角处中断.
Changes and Roles of Dorsal Root Ganglia in Nav1.7-related Congenital Insensitivity to Pain
Objective To investigate the changes of dorsal root ganglia(DRG)neurons in rats with voltage-gated sodium channel 1.7 loss-of-function(Nav1.7 LOF)and its involvement in congenital insensitivity to pain.Methods Immunofluorescence,Western blot,and alternative splicing analysis were used to verify the expression deletion of DRG neurons and Nav1.7 in spinal cord of Nav1.7 LOF rats.Using antibodies for peripherin,calcitonin gene-related peptide(CGRP),isolectin B4(IB4),and neuronal nuclear antigen(NeuN)in the DRG and spinal cord tissues,immunofluorescence staining was performed to observe the differences in DRG neuronal sub-populations and central terminal projections between Nav1.7 LOF rats and wild-type rats.Bulk RNA-Seq and real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the difference of gene expression in the DRG neurons between Nav1.7 LOF and WT rats.Following 55℃ thermal stimulation of the hind paw in Nav1.7 LOF rats,c-Fos and phospho-extracellular signal-regulated kinase(p-ERK)antibody staining was used to observe the activation of DRG neurons and spinal dorsal horn neurons.Results Nav1.7 was absent in the DRG neurons and spinal cord of Nav1.7 LOF rats.Compared with wild-type rats,Nav1.7 LOF rats exhibited a decreased proportion of IB4+non-peptidergic neurons in DRG tissues.Additionally,there was a significant loss of central projections of IB4+neurons in the spinal dorsal horn.The RNA-Seq and RT-qPCR results demonstrate that in Nav1.7 LOF rats,the gene expression of Mas-related G protein-coupled receptor(MRGPR),markers for 1B4+non-peptidergic neurons,was downregulat-ed.Compared with WT rats,there was no difference in c-Fos and p-ERK staining in DRG neurons of Nav1.7 LOF rats following ther-mal stimulation of the hindpaw.However,there was no c-Fos or p-ERK staining in spinal dorsal horn neurons,suggesting that the neu-rons were not activated.Conclusion Nav1.7 LOF rats exhibit a decreased proportion of IB4+non-peptideric neurons in DRG tissue and their central terminal projections.Noxious stimuli could activate DRG neurons,but could not activate dorsal horn neurons,suggesting the pain signal is interrupted between the central terminals of DRG neurons and the spinal dorsal horn.

Nav1.7Congenital insensitivity to painDorsal root gangliaNon-peptidergic neuron

赵廷玉、潘铭、施金叶、王爱忠、陈鲁滨、周全红

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200233 上海交通大学医学院附属第六人民医院麻醉科

201306 上海海洋大学水产与生命学院

Nav1.7 先天性无痛症 背根神经节 非肽能神经元

2024

医学研究杂志
中国医学科学院

医学研究杂志

CSTPCD
影响因子:0.702
ISSN:1673-548X
年,卷(期):2024.53(12)