摘要
目的研究lncRNA SNHG20在神经元活化过程中的作用。方法通过对KCl激活的神经元数据库进行分析,筛选出变化明显的lncRNA;使用实时荧光定量PCR(RT-qPCR)对在体外培养的KCl活化的神经元以及学习记忆模式刺激的实验动物的海马脑区进行结果验证;RT-qPCR以及Western blot检测过表达SNHG20的神经元中相关基因的表达;使用4-Thiouridine(4sU)标记新生成的RNA并通过RT-qPCR检测新生成的c-Fos mRNA水平;海马脑区AAV病毒脑注射后,对小鼠进行相关的行为学检测。结果SNHG20的表达水平在神经元活化后快速并且明显下降。SNHG20过表达后,抑制了早期响应基因c-Fos的RNA以及蛋白质水平,且新生成的c-Fos的mRNA水平较对照组明显下降。SNHG20也抑制了c-Fos通路下游靶基因,特别是Annexin A2(ANXA2)的水平。在体的海马脑区SNHG20过表达的小鼠表现出明显的学习记忆功能障碍。结论SNHG20能够通过抑制c-Fos的表达参与调控学习记忆过程。
Aim To investigate the effect of lncRNA SNHG20 on activation of neurons.Methods The lncRNA with significant level changes were screened by analyzing the database of KCl-activated neurons;the results were verified by real-time quantitative PCR(RT-qPCR)in KCl-activated neurons cultured in vitro and in the hippocampus of mice that were subjected to learning and memory training paradigm;the expression levels of related genes were examined by RT-qPCR and Western blot in SNHG20-administered neurons;nascent c-Fos mRNA levels were determined by RT-qPCR in 4-Thiouridine(4sU)labeled nascent RNA;behavioral tests were conducted on wild-type mice with AAV virus injected into the hippocampus of mouse brains.Results The levels of SNHG20 decreased rapidly and significantly after neuronal activation.After overexpression of SNHG20,both RNA and protein levels of the immediate early gene c-Fos were suppressed,and the levels of newly synthesized c-Fos mRNA were significantly lower than those of controls.SNHG20 was also involved in the regulation of c-Fos downstream genes,especially Annexin A2(ANXA2).Administration of SNHG20 in the hippocampus of mouse brain led to significant learning and memory deficits.Conclusion SNHG20 modulates learning and memory by inhibiting the expression of c-Fos.
作者
梅颖
李哓会
李定丰
张娟
刘强
MEI Ying;LI Xiao-hui;LI Ding-feng;ZHANG Juan;LIU Qiang(The First Affiliated Hospital of USTC,School of Life Sciences,Division of Life Sciences and Medicine,University of Science and Technology of China,Hefei 230001,China;Institute on Aging and Brain Disorders,the First Affiliated Hospital of USTC,University of Science and Technology of China,Hefei 230001,China;Neurodegenerative Disease Research Center,University of Science and Technology of China,Hefei 230026,China;CAS Key Lab of Brain Function and Disease,University of Science and Technology of China,Hefei 230026,China;National Synchrotron Radiation Lab,University of Science and Technology of China,Hefei 230026,China;CAS Center for Excellence in Animal Evolution and Genetics,Chinese Academy of Sciences,Kunming 650223,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2020年第11期1600-1606,共7页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 31871082)。
