摘要
目的:评价海马齿状回(DG)区钠离子渗漏通道蛋白(NALCN)与小鼠认知功能的关系。方法:雄性野生型C57BL/6小鼠39只,6~8周龄,体重18~22 g。取3只小鼠采用免疫荧光染色法检测海马DG区NALCN和神经元核抗原(NeuN)共定位表达。余36只小鼠采用随机数字表法分为2组(n=18):对照组(C组)和NALCN基因敲除组(KO组)。KO组于海马DG区注射NALCN腺相关病毒,C组注射对照病毒。病毒注射后3周,进行行为学测试(Y迷宫实验和旷场实验),随后处死,取海马组织,分别采用Western blot法和RT-PCR法检测海马DG区NALCN及其mRNA表达。结果:小鼠海马DG区同一神经元NALCN和NeuN有大量共定位,NALCN在海马DG区广泛表达。与C组比较,KO组海马DG区NALCN及其mRNA表达下调,进入新臂次数减少,新臂停留时间缩短(P<0.05),旷场实验各指标差异无统计学意义(P>0.05)。结论:海马DG区NALCN参与小鼠认知功能的调控,其表达下调可导致认知功能减退。
Objective To evaluate the relationship between Na+leak current channel(NALCN)in hippocampal dentate gyrus(DG)region and cognitive function in mice.Methods Thirty-nine male wild type C57BL/6 mice,aged 6-8 weeks,weighing 18-22 g,were studied.Three mice were sacrificed to verify the expression of NALCN co-localized with neuronal nuclear antigen(NeuN)in the hippocampal DG region through immunofluorescence technique.The remaining 36 mice were divided into 2 groups(n=18 each)by the random number table method:control group(group C)and NALCN gene knockout group(group KO).NALCN-shRNA virus was injected in hippocampal DG region in group KO,and scrambled-shRNA virus was injected in group C.Three weeks after virus injection,behavioral tests(Y maze test and open field test)were performed,then the animals were sacrificed,and the hippocampal tissues were removed for determination of the expression of NALCN protein and mRNA using Western blot and real-time polymerase chain reaction.Results NALCN and NeuN colocalized a lot on the same neuron in the hippocampal DG region of mice,and NALCN was widely expressed in the hippocampal DG region.Compared with group C,the expression of NALCN protein and mRNA was significantly down-regulated,the times of entering the new arm were reduced,the duration of staying at the new arm was shortened(P<0.05),and no significant change was found in the parameters mentioned above in the open field test in group KO(P>0.05).Conclusions NALCN in the hippocampal DG region is involved in the regulation of cognitive function in mice,and the down-regulation of NALCN may lead to cognitive decline.
作者
赵军博
耿晓媛
王丽娟
张晓艳
姜丽华
Zhao Junbo;Geng Xiaoyuan;Wang Lijuan;Zhang Xiaoyan;Jiang Lihua(Department of Anesthesiology,The Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2022年第12期1461-1464,共4页
Chinese Journal of Anesthesiology
基金
河南省医学科技攻关计划联合共建项目(2018020195)。
关键词
齿状回
钠通道
认知功能障碍
Dentate gyrus
Sodium channels
Cognitive dysfunction
