Mechanical Stretch Promotes Neurite Outgrowth of Primary Cultured Dorsal Root Ganglion Neurons via Suppression of Semaphorin 3A-Neuropilin-1/Plexin-A1 Signaling
Mechanical Stretch Promotes Neurite Outgrowth of Primary Cultured Dorsal Root Ganglion Neurons via Suppression of Semaphorin 3A-Neuropilin-1/Plexin-A1 Signaling
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机械拉伸通过抑制 Semaphorin 3A-Neuropilin-1/Plexin-A1 信号传导促进原代培养的背根神经节神经元的神经突生长
DOI：
10.1021/acschemneuro.2c00432
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发表时间：
2022
影响因子：
5
通讯作者：
Yubo Fan
中科院分区：
文献类型：
--
作者：
Meili Liu;Zitong An;Yu Zhang;Yuchen Xiao;Junwei Xu;Zhijun Zhao;Chongquan Huang;Anqing Wang;Gang Zhou;Ping Li;Yubo Fan
在神经发育和再生中，已经进行了大量的尝试来促进神经元的延伸和迁移。虽然机械拉伸诱导轴突的持续伸长，但其潜在的分子机制尚不清楚。生长锥内分泌的一些轴突导向因子影响轴突的生长，在轴突连接中对轴突有吸引或排斥作用。Semaphorin 3A（Sema 3A）是一种重要的排斥导向分子，抑制Sema 3A的表达可以促进神经元的发育。在这项研究中，机械拉伸对背根神经节神经元生长的影响和潜在的机制进行了研究，通过评估的延伸方向，轴突长度，细胞体的大小，线粒体膜电位，和Sema 3A及其受体的表达。我们的研究结果表明，细胞活力显着增加，在拉伸应变为2.5%，5%和10%的4小时，最突出的效果在5%的拉伸应变。在5%的拉伸应变下（0-12 h），神经元向拉伸方向迁移，而对照组神经元的迁移方向是无序的。此外，Sema 3A-神经纤毛蛋白-1/丛蛋白-A1信号通路被抑制后，机械拉伸在5%的拉伸应变4小时，通过免疫荧光染色，免疫沉淀，和蛋白质印迹分析。最后，Sema 3A-SiRNA（SiRNA =小干扰RNA）处理导致拉伸生长神经元的显著引导生长。更重要的是，机械拉伸处理后，排斥因子Sema 3A的表达显著降低，吸引分子Netrin-1的表达显著增加，这共同促进了神经突的生长。本研究为机械牵张疗法或导向因子相关药物在损伤神经元再生中的应用提供了新的思路。
Significant attempts have been made to promote neuronal extension and migration in nerve development and regeneration. Although mechanical stretch induces persistent elongation of the axon, the underlying molecular mechanisms are not yet clear. Some axonal guidance cues secreted in the growth cone that affect the axonal growth could attract or repel axons in neurite connection. As semaphorin 3A (Sema3A) is an important repulsion guidance molecule, inhibition of Sema3A has been postulated to promote neuronal development. In this study, the effects of mechanical stretch on dorsal root ganglion neuronal growth and the underlying mechanisms were investigated by assessing the extension direction, neurite length, cell body size, mitochondrial membrane potential, and the expression of Sema3A and its receptors. Our results showed that cell viability significantly increased at tensile strains of 2.5, 5, and 10% for 4 h, with the most prominent effect at 5% tensile strain. Moreover, neurons migrated closer to the stretching direction at 5% tensile strain (0–12 h), while the neurons of the control group moved in a disorderly manner. Furthermore, Sema3A-Neuropilin-1/Plexin-A1 signaling pathway was found to be suppressed after mechanical stretch at 5% tensile strain for 4 h by immunofluorescence staining, immunoprecipitation, and western blot assay. Finally, a Sema3A-SiRNA (SiRNA = small interfering RNA) treatment led to remarkable guidance growth in the stretch-grown neurons. Importantly, there was significant decrease of repulsive cue Sema3A expression and remarkable increase of attractive molecule Netrin-1 expression after mechanical stretching treatment, which jointly promoted neurite outgrowth. This study provides a promising new approach for the development of mechanical stretching therapy or guidance factor-related drugs in injured neuronal regeneration.