1,800 MHz Radiofrequency Electromagnetic Irradiation Impairs Neurite Outgrowth With a Decrease in Rap1-GTP in Primary Mouse Hippocampal Neurons and Neuro2a Cells.
1,800 MHz Radiofrequency Electromagnetic Irradiation Impairs Neurite Outgrowth With a Decrease in Rap1-GTP in Primary Mouse Hippocampal Neurons and Neuro2a Cells.
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1,800 MHz 射频电磁辐射会损害原代小鼠海马神经元和 Neuro2a 细胞中 Rap1-GTP 的减少，从而损害神经突生长
DOI：
10.3389/fpubh.2021.771508
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发表时间：
2021
影响因子：
5.2
通讯作者：
Zhang L
中科院分区：
文献类型：
--
作者：
Li Y;Deng P;Chen C;Ma Q;Pi H;He M;Lu Y;Gao P;Zhou C;He Z;Zhang Y;Yu Z;Zhang L
背景：随着移动电话等通信设备在全球的普及，人们越来越关注射频电磁辐射(RF-EMR)对大脑的影响，大脑是对1800 MHz射频电磁辐射最敏感的器官之一。然而，RF-EMR暴露对神经细胞的影响尚不清楚。轴突生长在脑发育中起着至关重要的作用，因此，确定1800 MHz射频电磁辐射对轴突生长的影响对于探索其对脑发育的影响具有重要意义。目的：研究1800 MHz射频电磁辐射暴露48h对神经细胞突起生长的影响，并探讨RAP1信号通路的相关作用。材料和方法：将C57BL/6小鼠原代海马神经元和Neuro2a细胞以4W/kg的比吸收率(SAR)1800 MHz RF-EMR照射48h，用CCK-8比色法测定照射后24、48、72h的细胞存活率。用20倍光学显微镜观察原代海马神经元(DIV-2)和Neuro2a细胞的突起生长，并用ImageJ软件进行识别。实时荧光定量聚合酶链式反应检测Rap1a和Rap1b基因的表达。免疫印迹法检测RAP1、RAP1a、RAP1b、RAP1GAP和p-MEK1/2蛋白表达。免疫沉淀法检测RAP1-GTP的表达。通过将RAP1-Gly_Val-GFP基因导入Neuro2a细胞，评价了RAP1-GTP的作用。结果：4W/kg 1800MHzRF-EMR作用24、48、72h对细胞活力无明显影响。48h RF-EMR暴露可显著损伤原代培养小鼠海马神经元的轴突长度、初级和次级突起数目以及分支点。48h的RF-EMR对Neuro2a细胞的轴突细胞百分率和轴突长度也有抑制作用。48h RF-EMR可抑制RAP1的活性，但对RAP1的基因和蛋白表达均无明显影响。RF-EMR暴露48h后，Rap1GAP蛋白表达增加，而p-MEK1/2蛋白表达降低。RAP1的过表达逆转了1800 MHz射频电磁辐射48h诱导的神经细胞RAP1-GTP的降低和突起生长损伤。结论：RAP1活性及其相关信号通路参与了1800 MHz RF-EMR暴露48h对神经细胞突起生长的影响。RF-EMR暴露对婴儿和儿童神经元发育的影响值得更多关注。
Background: With the global popularity of communication devices such as mobile phones, there are increasing concerns regarding the effect of radiofrequency electromagnetic radiation (RF-EMR) on the brain, one of the most important organs sensitive to RF-EMR exposure at 1,800 MHz. However, the effects of RF-EMR exposure on neuronal cells are unclear. Neurite outgrowth plays a critical role in brain development, therefore, determining the effects of 1,800 MHz RF-EMR exposure on neurite outgrowth is important for exploring its effects on brain development. Objectives: We aimed to investigate the effects of 1,800 MHz RF-EMR exposure for 48 h on neurite outgrowth in neuronal cells and to explore the associated role of the Rap1 signaling pathway. Material and Methods: Primary hippocampal neurons from C57BL/6 mice and Neuro2a cells were exposed to 1,800 MHz RF-EMR at a specific absorption rate (SAR) value of 4 W/kg for 48 h. CCK-8 assays were used to determine the cell viability after 24, 48, and 72 h of irradiation. Neurite outgrowth of primary hippocampal neurons (DIV 2) and Neuro2a cells was observed with a 20 × optical microscope and recognized by ImageJ software. Rap1a and Rap1b gene expressions were detected by real-time quantitative PCR. Rap1, Rap1a, Rap1b, Rap1GAP, and p-MEK1/2 protein expressions were detected by western blot. Rap1-GTP expression was detected by immunoprecipitation. The role of Rap1-GTP was assessed by transfecting a constitutively active mutant plasmid (Rap1-Gly_Val-GFP) into Neuro2a cells. Results: Exposure to 1,800 MHz RF-EMR for 24, 48, and 72 h at 4 W/kg did not influence cell viability. The neurite length, primary and secondary neurite numbers, and branch points of primary mouse hippocampal neurons were significantly impaired by 48-h RF-EMR exposure. The neurite-bearing cell percentage and neurite length of Neuro2a cells were also inhibited by 48-h RF-EMR exposure. Rap1 activity was inhibited by 48-h RF-EMR with no detectable alteration in either gene or protein expression of Rap1. The protein expression of Rap1GAP increased after 48-h RF-EMR exposure, while the expression of p-MEK1/2 protein decreased. Overexpression of constitutively active Rap1 reversed the decrease in Rap1-GTP and the neurite outgrowth impairment in Neuro2a cells induced by 1,800 MHz RF-EMR exposure for 48 h. Conclusion: Rap1 activity and related signaling pathways are involved in the disturbance of neurite outgrowth induced by 48-h 1,800 MHz RF-EMR exposure. The effects of RF-EMR exposure on neuronal development in infants and children deserve greater focus.
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Rap1 GTPase的许多面孔。
影响因子：
5.6
作者：
Jaśkiewicz A;Pająk B;Orzechowski A
通讯作者：
Orzechowski A
DOI：
10.3390/ijerph17249179
发表时间：
2020-12-08
影响因子：
--
作者：
Ishihara T;Yamazaki K;Araki A;Teraoka Y;Tamura N;Hikage T;Omiya M;Mizuta M;Kishi R
通讯作者：
Kishi R
影响因子：
4.7
作者：
Baron J;Blex C;Rohrbeck A;Rachakonda SK;Birnbaumer L;Ahnert-Hilger G;Brunk I
通讯作者：
Brunk I
影响因子：
2.5
作者：
Inoue, Shigeki;Motoda, Hirotoshi;Kano, Yoshio
通讯作者：
Kano, Yoshio
影响因子：
7.7
作者：
Araki, Yoichi;Hong, Ingie;Huganir, Richard L.
通讯作者：
Huganir, Richard L.