中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (7): 1535-1544.doi: 10.4103/1673-5374.330615

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

光遗传学技术刺激缺血性卒中小鼠对侧前外侧运动皮质区初级运动皮质神经的重组

  

  • 出版日期:2022-07-15 发布日期:2022-01-17

Optogenetics stimulates nerve reorganization in the contralesional anterolateral primary motor cortex in a mouse model of ischemic stroke

Bei-Yao Gao1, #, Yi-Xing Cao2, #, Peng-Fei Fu3, Ying Xing4, Dan Liang4, Shan Jiang1,Yu-Xiao Xie1, *, Min Li2, *#br#   

  1. 1Department of Rehabilitation Medicine, China-Japan Friendship Hospital, Beijing, China; 2Institute of Future Lighting, Academy for Engineering and Technology, Fudan University, Shanghai, China; 3Department of Neurosurgery, Huashan Hospital, Fudan University, Shanghai, China; 4Department of Rehabilitation, Huashan Hospital, Fudan University, Shanghai, China
  • Online:2022-07-15 Published:2022-01-17
  • Contact: Min Li, PhD, lm_ss@fudan.edu.cn; Yu-Xiao Xie, MD, 13501073965@163.com.

摘要:

啮齿类动物前外侧运动皮质是重要的运动辅助区之一,其功能与人类的运动前区相近。有研究认为前外侧运动皮质的激活和抑制对运动的行为有直接的影响,但其在脑卒中的治疗意义仍然不明确。实验观察了光抑制和光激活对侧前外侧运动皮质在脑卒中小鼠模型中的作用。发现21d的光遗传学刺激对侧前外侧运动皮质可改善模型小鼠神经功能,且只有光抑制有这种作用。进一步观察可见光抑制和光激活后树突状结构重组、树突状棘可塑性等存在显著差异:其中经光抑制后,缺血侧初级运动皮质的树突长度、树突棘数目和穿孔突触数量均增加,而经光激活后,则可见缺血侧初级运动皮质中多个突触钮扣数量和树突交叉点增加。且RNA测序结果显示,光抑制后即刻,生物过程的正调控被上调,其中cFOS、Erg1、Sema3f等基因的富集大于光激活,这一结果已经实时定量逆转录聚合酶链式反应验证。同时免疫荧光结果也显示,与光激活相比,光抑制后缺血侧初级运动皮质V层中c-FOS荧光阳性反应增强。结果表明,光遗传学刺激对侧前外侧运动皮质对脑卒中后缺血侧初级运动皮质中神经重组是有益的,且光激活和光抑制对神经可塑性的影响是不同的。实验于2018年3月3日经复旦大学动物伦理委员会批准(批准号201802173S)。

https://orcid.org/0000-0002-7850-3383 (Min Li); https://orcid.org/0000-0002-9335-9017 (Yu-Xiao Xie)

关键词:

脑卒中, 光遗传学, 前外侧运动皮质, 运动功能, 大脑中动脉闭塞, 神经可塑性, 运动活动, 突触

Abstract: The anterolateral motor cortex of rodents is an important motor auxiliary area, and its function is similar to that of the premotor area in humans. Activation and inhibition of the contralesional anterolateral motor cortex (cALM) have been shown to have direct effects on motor behavior. However, the significance of cALM activation and inhibition in the treatment of stroke remains unclear. This study investigated the role of optogenetic cALM stimulation in a mouse model of cerebral stroke. The results showed that 21-day optogenetic cALM inhibition, but not activation, improved neurological function. In addition, optogenetic cALM stimulation substantially altered dendritic structural reorganization and dendritic spine plasticity, as optogenetic cALM inhibition resulted in increased dendritic length, number of dendritic spines, and number of perforated synapses, whereas optogenetic activation led to an increase in the number of multiple synapse boutons and the number of dendritic intersections. Furthermore, RNA-seq analysis showed that multiple biological processes regulated by the cALM were upregulated immediately after optogenetic cALM inhibition, and that several immediate-early genes (including cFOS, Erg1, and Sema3f) were expressed at higher levels after optogenetic inhibition than after optogenetic activation. These results were confirmed by quantitative reverse transcription-polymerase chain reaction. Finally, immunofluorescence analysis showed that the c-FOS signal in layer V of the primary motor cortex in the ischemic hemisphere was higher after optogenetic cALM activation than it was after optogenetic cALM inhibition. Taken together, these findings suggest that optogenetic cALM stimulation promotes neural reorganization in the primary motor cortex of the ischemic hemisphere, and that optogenetic cALM inhibition and activation have different effects on neural plasticity. The study was approved by the Experimental Animal Ethics Committee of Fudan University (approval No. 201802173S) on March 3, 2018.

Key words: anterolateral motor cortex, locomotion activity, middle cerebral artery occlusion, motor function, neural plasticity, optogenetics, stroke, synapse