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Nature文献jie读-mRNAm6A甲ji化调控对学xi记忆功能机謕in挠跋
点击次数:2428 发布日期:2021-1-4  来源:本站 仅供参考,谢绝转载,否则责任自fu
学xi和记忆是大脑的ji本功能,长shi记忆的形成bei认weixu要神经元活动诱导xia的蛋白质合成。以往的研jiu表明,m6A参与调节神经功能,包括小shu中脑多巴胺能信hao传导、飞虫飞xingxingwei、成nian小shu神经生成和轴tuzai生。在大脑发育、xingwei体验和记忆形成过cheng中观察daom6A祅a系鳎獗砻鱩6A的积累和大脑活动zhi间cun在相guan性。而YTHDFjiazu蛋白ke以与mRNA上的m6A结合,从而调控mRNA的翻译,同shi前期实验已证实YTDHF1在小shu海马体中高表da。因此也提示了学xi和记忆过chengke能hui受daom6A及qi结合蛋白Ythdf1翻译效ying的影响。
 
研jiu团dui利用CRISPR/Cas9技术,培育出了Ythdf1 敲chu小shu (Ythdf1-KO)。Morris水迷宫实验显示与ye生型小shu对bi,Ythdf1-KO小shu的空间记忆能力明显jiang低。
 
另外通过经dian的条件恐惧测试实验,在经过电击刺激的训lianzhihou,Ythdf1-KO小shu在实验间xie期(ITI)freezing的表现相对较少,而ting觉相guan的恐惧反ying(与杏仁核功能相guan)(即freezing)的表现没有改bian,由此也提示了 Ythdf1的缺失扰乱了小shu海马正chang的空间学xi记忆能力。具体实验数据如xia图所示。
 
 
Figure 1.Impaired spatial learning and memory in Ythdf1-KO mice.
ab, Representative images of Ythdf1 immunostaining (a) and Hoechst (b) in the control and Ythdf1-KO hippocampus. DG, dentate gyrus; P30/P120, postnatal day 30/120. cd, Learning curves of control (blue) and Ythdf1-KO (red) mice in Morris water maze (MWM) tests in visible (c) and hidden (d) platform training. e, Quadrant time (%) (left) and representative swimming paths (right) of control and Ythdf1-KO mice in the MWM probe test. The red dash line represents the chance level (25%). f, Learning curves of control (blue) and Ythdf1-KO mice (red) for contextual fear conditioning (FC) in moderate (left) or strong (right) training sessions. Base, baseline; ITI, inter-trial interval. gh, Contextual fear memory assessed 24 hours (g) or 2 hours (h) after the indicated FC. P values, two-way ANOVA with two tailed t-test (relative to “Target” or between genotypes) (e), two-way repeated measures ANOVA with post hoc test (cdf), and two-tailed t-test (gh). Numbers in bars, numbers of mice. Error bars, mean ± s.e.m.
 
wei了tanjiuYthdf1的作用机制,通过电生理实验,研jiu团dui发现相较于对照组,Ythdf1-KO小shu CA1神经元的mEPSCs的振fu和pinlv显zhujiang低 (Fig. 2a-b)。配对mai冲bi(PPRs)分析也显示Ythdf1-KO CA1神经元tuchu前shi放概lvjiang低((Extended Data Fig. 4a-b),),由此也证实了Ythdf1缺失造成了ji础tuchu传递功能的缺陷。同shi形态学染色结果显示,Ythdf1-KO CA1神经元jiang低了树tuji密度,danbing未改bianji的大小 (Extended Data Fig. 4c-d)。
 
 
Figure 2. Deficient basal transmission and plasticity in Ythdf1-KO hippocampal synapses.
a,b, Representative traces (a) and quantification of amplitude (b, left) and frequency (b, right) of spontaneous miniature excitatory postsynaptic currents (mEPSCs) in control and Ythdf1-KO hippocampal CA1 neurons. c, d, Summary plots (c) and average amplitude (d) of long-term potentiation (LTP) induced by 2 × high frequency stimulation (HFS) in the CA1 region of control and Ythdf1-KO acute slices. fEPSP, field excitatory postsynaptic potential. e, f, Summary plots (e) and average amplitude (f) of late phase LTP induced by 4 × HFS. Top panels, sample traces taken at time points 1 and 2 indicated above the summary plots; scale bars, 10 ms (horizontal) and 0.2 mV (vertical) (c, e). g, h, Representative western blots (g) and quantification (h) of a number of LTP-related proteins in the control and Ythdf1-KO hippocampal postsynaptic density (PSD) fraction. P values, Kolmogorov-Smirnov test for cumulative distributions followed by comparisons with Mann–Whitney U test (b) and two-tailed t-test (d, f, h). Numbers in bars, numbers of neurons/mice (b), slices/mice (d, f), and mice (h). Error bars, mean ± s.e.m
 
wei了进一步tanjiuYthdf1是否能够调控长shicheng的tuchuke塑性,通过一种zhu流的阐shi学xi记忆的分zi机制——长shicheng增强效ying(Long Term Potentiation ,LTP),即通过给与海马CA1区高pin刺激,记录Ythdf1-KO和对照组小shu(ye生型)的LTP,结果显示,Ythdf1-KO未能显示正chang水ping的tuchuhou电位。同shiYthdf1的缺失显zhujiang低了海马神经元tuchuhouzhi密物中参与LTP的guan键蛋白质的水ping (Fig. 2g-h; Extended Data Fig. 4e)。以上实验结果显示,Ythdf1的缺失损害了海马区神经元的tuchu传导功能,由磗huo紌hi学xi和记忆能力的xiajiang。
 
 
Extended Data Figure 4 |. Paired-pulse ratios (PPR), spine morphology, and total protein levels of various LTP-related genes in Ythdf1-KO mouse hippocampus, related to Figure 2.
a, b, PPR with different inter-stimulus intervals in CA1 neurons from wild-type control and Ythdf1-KO mice. c, d, Representative images of Lucifer Yellow staining (c) and statistical analyses of spine density (d, left) and spine size (d, right) in CA1 neurons from adult control and Ythdf1-KO brain. e, Uncropped western blot images for Fig. 2g. f, Total protein levels of a set of LTP-related genes in control and Ythdf1-KO mouse hippocampus. For gel source data, see Supplementary Figure 1. P values, two-way repeated measures ANOVA with post hoc two-tailed t-test (a) and two tailed t-test (b, d, f). Numbers in bars, numbers of slices (b), neurons/mice (d, left), spines (d, right), or mice (f). Error bars, mean ± s.e.m.
 
wei了证实小shu学xi记忆功能的xiajiang是由海马区Ythdf1缺失导zhi的,研jiu团dui针对Ythdf1-KO成nian小shu重表da了海马区Ythdf1,Morris水迷宫实验和情景相guan条件恐惧实验证实,重表daYthdf1的Ythdf1-KO小shu学xi记忆能力大fu度提高。同shiKCl去极化刺激实验和电痉挛疗法实验显示, Ythdf1介导的蛋白质合成功能,ke在神经元的刺激xia趋于正chang化。
 
此项研jiu将m6A甲ji化这一生物体普biancun在的修饰机制研jiu引入daojie决神经科学领域guan键科学问ti中,对于kai启和深入阐明蛋白翻译调控在tuchuke塑性中的作用有zhou重要的意义。
 
就记忆功能而言,ke分weiduanshi(工作记忆)和长shi记忆。Anderson(1976)将长shi记忆分wei砽v鲂约且浠蚍浅lv鲂约且洹3lv鲂约且渖婕暗膠hu要脑结gou是海马以及qi他颞叶内ce结gou。非砽v鲂约且鋍hun成了yun动技能和xi惯的获得,bing通过新纹状体和小脑实现。此外,杏仁核介导情xu记忆,bingbei证明参与记忆的整合。
 
伴随zhou脑科学研jiu的buduan深入,xingwei学已经是必buke少的研jiushou段。
 
来源:深圳市瑞沃德生命科技有限公司
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