中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (12): 2147-2155.doi: 10.4103/1673-5374.262593

• 原著:退行性病与再生 • 上一篇    下一篇

光感基因神经调控技术激活谷氨酸受体改善阿尔茨海默病小鼠的记忆功能

  

  • 出版日期:2019-12-15 发布日期:2019-12-15
  • 基金资助:

    中国国家自然科学基金项目（81171191），深圳市战略性新兴产业发展专项基金项目（JCYJ20160422170522075），深圳医疗健康研究项目（201601015）

Optogenetics-induced activation of glutamate receptors improves memory function in mice with Alzheimer’s disease

Ke-Wei Wang 1 , Xiao-Lin Ye 1 , Ting Huang 1 , Xi-Fei Yang 2 , Liang-Yu Zou 1   

  1. 1 Department of Neurology, Shenzhen People’s Hospital (First Affiliated Hospital of Southern University of Science and Technology), Second Clinical College, Jinan University, Shenzhen, Guangdong Province, China
    2 Key Laboratory of Modern Toxicology of Shenzhen, Shenzhen Center for Disease Control and Prevention, Shenzhen, Guangdong Province, China
  • Online:2019-12-15 Published:2019-12-15
  • Contact: Liang-Yu Zou, PhD, MD, zouliangyu@yahoo.com; Xi-Fei Yang, PhD, xifeiyang@gmail.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81171191 (to LYZ); the Shenzhen Special Fund Project on Strategic Emerging Industry Development of China, No. JCYJ20160422170522075 (to LYZ); the Shenzhen Healthcare Research Project of China, No. 201601015 (to LYZ).

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摘要：

光感基因神经调控技术是一种结合光学和遗传学的技术，可激活或抑制活组织以及活体动物中特定的细胞，已被用干预帕金森病、癫痫以及神经疾病，但是光感基因神经调控技术治疗阿尔茨海默病的研究。(1)实验将携带光敏感通道蛋白2的腺相关病毒被注射到C57BL/6小鼠双侧海马齿状回中，同时反复在海马内注射可溶性低分子量Aβ1-42同时以473nm激光（1-3ms，10Hz，5min）进行刺激；(2)以新奇物识别实验测试小鼠的记忆功能，以免疫组化分析小鼠海马组织中神经元神经胶质细胞NeuN和突触素Ia/b的免疫阳性细胞的数量，以Western blot分析海马组织中神经胶质细胞特异性蛋白胶质纤维酸性蛋白、NeuN、突触素Ia/b、谷氨酸受体相关蛋白代谢型谷氨酸受体1a、代谢型谷氨酸受体5、代谢型谷氨酸受体亚基NR1和谷氨酸受体2以及神经炎性因子白细胞介素1β、白细胞介素6以及白细胞介素10的表达水平；(3)结果显示，应用光感基因神经调控技术可改善阿尔茨海默病小鼠的工作记忆和短期记忆，增加海马谷氨酸受体代谢型谷氨酸受体亚基NR1、谷氨酸受体2与代谢型谷氨酸受体5表达增加，抑制胶质纤维酸性蛋白和白细胞介素6的表达，而其他蛋白表达或免疫阳性反应无明显变化；(4)实验结果提示，光感基因神经调控技术可精准和特异性激活谷氨酸能神经元，调节谷氨酸受体，抑制炎症反应，从而调节神经元-神经胶质细胞网络，改善阿尔茨海默病小鼠的工作记忆和短期记忆功能。实验于2011年2月28日获得暨南大学动物资源委员会的批准（批准号LL-KT-2011134）。

orcid: 0000-0001-8066-8892 (Liang-Yu Zou)

关键词: 阿尔茨海默病, A&beta, 1-42, 海马齿状回, 光敏感通道蛋白2, 谷氨酸受体, 记忆, 神经炎症, 新物体识别, 免疫组化, Western blot分析, 神经再生

Abstract:

Optogenetics is a combination of optics and genetics technology that can be used to activate or inhibit specific cells in tissues. It has been used to treat Parkinson’s disease, epilepsy and neurological diseases, but rarely Alzheimer’s disease. Adeno-associated virus carrying the CaMK promoter driving the optogenetic channelrhodopsin-2 (CHR2) gene (or without the CHR2 gene, as control) was injected into the bilateral dentate gyri, followed by repeated intrahippocampal injections of soluble low-molecular-weight amyloid-β1–42 peptide (Aβ1–42). Subsequently, the region was stimulated with a 473 nm laser (1–3 ms, 10 Hz, 5 minutes). The novel object recognition test was conducted to test memory function in mice. Immunohistochemical staining was performed to analyze the numbers of NeuN and synapsin Ia/b-positive cells in the hippocampus. Western blot assay was carried out to analyze the expression levels of glial fibrillary acidic protein, NeuN, synapsin Ia/b, metabotropic glutamate receptor-1a (mGluR-1a), mGluR-5, N-methyl-D-aspartate receptor subunit NR1, glutamate receptor 2, interleukin-1β, interleukin-6 and interleukin-10. Optogenetic stimulation improved working and short-term memory in mice with Alzheimer’s disease. This neuroprotective effect was associated with increased expression of NR1, glutamate receptor 2 and mGluR-5 in the hippocampus, and decreased expression of glial fibrillary acidic protein and interleukin-6. Our results show that optogenetics can be used to regulate the neuronal-glial network to ameliorate memory functions in mice with Alzheimer’s disease. The study was approved by the Animal Resources Committee of Jinan University, China (approval No. LL-KT-2011134) on February 28, 2011.

Key words: nerve regeneration, Alzheimer’s disease, amyloid-β1–42, dentate gyrus, channelrhodopsin-2, glutamate receptors, memory, neuroinflammation, novel object recognition, immunohistochemistry, western blot assay, neural regeneration