目的 利用泛素羧基末端水解酶L5(UCHL5)脑部基因敲除小鼠模型,研究UCHL5对空间学习记忆功能的影响.方法 使用CRISPR/Cas9技术构建UCHL5脑部条件敲除小鼠.选择4月龄及以上的UCHL5脑敲除小鼠与野生型小鼠各16只.他莫昔芬腹腔注射连续5次,隔日1次,建模成功后采用Western blotting方法鉴定小鼠脑组织UCHL5表达水平.利用Morris水迷宫实验检测UCHL5敲除对小鼠空间学习记忆能力的影响.结果 Western blotting实验结果表明UCHL5蛋白在敲除小鼠脑组织中表达缺失.UCHL5敲除影响小鼠空间学习记忆能力,定位航行实验结果显示UCHL5脑敲除小鼠的平均潜伏期长于野生型小鼠,在训练第1、3日平均潜伏期延长,但差异无统计学意义(P=0.403,P=0.067),在训练第2、4、5日平均潜伏期均明显延长(均P＜0.05);空间探索实验结果显示UCHL5脑敲除小鼠在目标象限时间占比明显低于野生型小鼠[(7.71±0.81)％vs(9.60±0.51)％,P=0.042];UCHL5脑敲除小鼠的游泳总路程大于野生型小鼠[(7 500.92±14.35)cmvs(6 034.48±17.10)cm,P＜0.001].结论 UCHL5的表达水平影响小鼠空间学习记忆能力.
Abstract
Objective To investigate the effect of ubiquitin carboxyl-terminal hydrolyse L5(UCHL5)on spatial learning and memory function by UCHL5 knockout mouse models.Methods UCHL5 brain conditioned knockout mice were constructed using CRISPR/Cas9 technology.A total of 16 UCHL5 brain knockout mice and 16 wild-type mice aged 4 months and above were selected.Tamoxifen was injected intraperitoneally for 5 consecutive times,once every other day.After successful modeling,UCHL5 expression level in mouse brain tissue was identified by Western blotting method.Morris water maze experiment was used to detect the effect of UCHL5 knockout on the spatial learning and memory ability of mice.Results Western blotting experiments showed that the expression of UCHL5 protein was lost in the brain tissue of knockout mice.UCHL5 knockout affected the spatial learning and memory ability of mice.The results of place navigation test showed that the average latency of UCHL5 knockout mice was longer than that of wild-type mice,and the average latency was prolonged on the first and third day of training,but the difference was not statistically significant(P=0.403,P=0.067).The average latency was significantly prolonged on the 2nd,4th and 5th day of training(all P＜0.05).The results of space exploration experiments showed that the time ratio of UCHL5 knockout mice in the target quadrant was significantly lower than that of wild-type mice[(7.71±0.81)％vs(9.60±0.51)％,P=0.042].The total swimming distance of UCHL5 knockout mice was higher than that of wild-type mice[(7 500.92±14.35)cm vs(6 034.48±17.10)cm,P＜0.001].Conclusion UCHL5 expression level affects the spatial learning and memory ability of mice.
关键词
去泛素化酶/UCHL5/Morris水迷宫/空间学习记忆能力
Key words
deubiquitinase/UCHL5/Morris water maze/spatial learning and memory ability
维普
万方数据