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敲除MT3R/Nqo2基因对大鼠睡眠/觉醒行为和EEG能谱的影响
Effects of knockout of MT3R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats
- 陆军军医大学学报 2023年45卷第9期 页码：936-946
- 作者机构：
1.400038 重庆, 陆军军医大学(第三军医大学)基础医学院生理学教研室, 脑与智能研究院
2.857000 西藏 日喀则, 陆军第953医院急诊医学科
- 作者简介：
[何超，E-mail: hechaochongqing@163.comHE Chao, E-mail: hechaochongqing@163.com
- 基金信息：基金项目］ 国家自然科学基金面上项目(31970945);重庆市杰出青年科学基金项目(CSTB2022NSCQ-JQX0019);中国科技部科技创新2030-重大项目(2021ZD0203200-01)
DOI：10.16016/j.2097-0927.202212125
中图分类号： R-332;R338.63;R338.8收稿：2022-12-20，
修回：2023-02-03，
纸质出版：2023-05-15
- 稿件说明：
移动端阅览
李鑫, 李世音, 罗芬兰, 等. 敲除MT3R/Nqo2基因对大鼠睡眠/觉醒行为和EEG能谱的影响[J]. 陆军军医大学学报, 2023,45(9):936-946.
Xin LI, Shiyin LI, Fenlan LUO, et al. Effects of knockout of MT3R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats[J]. Journal of Army Medical University, 2023, 45(9): 936-946.
李鑫, 李世音, 罗芬兰, 等. 敲除MT3R/Nqo2基因对大鼠睡眠/觉醒行为和EEG能谱的影响[J]. 陆军军医大学学报, 2023,45(9):936-946. DOI： 10.16016/j.2097-0927.202212125.
Xin LI, Shiyin LI, Fenlan LUO, et al. Effects of knockout of MT3R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats[J]. Journal of Army Medical University, 2023, 45(9): 936-946. DOI： 10.16016/j.2097-0927.202212125.
摘要
目的
2
利用CRISPR/Cas9基因编辑工具构建褪黑素3型受体(MT
3
R/Nqo2)基因敲除的SD大鼠
观察基因敲除后大鼠的睡眠/觉醒行为变化和EEG能谱改变
以探究褪黑素调控睡眠/觉醒的受体机制。
方法
2
利用非同源重组修复引入突变的方式
造成MT
3
R/Nqo2基因蛋白读码框移码突变
使该基因功能缺失
建立MT
3
R/Nqo2基因敲除的SD大鼠
并通过PCR、RT-PCR和FISH进行基因水平、转录水平和形态学的敲除验证
然后对敲除组与对照组大鼠进行睡眠/觉醒行为分析和睡眠/觉醒各时相脑电能谱分析。
结果
2
MT
3
R/Nqo2基因敲除后
与对照组大鼠相比
敲除组大鼠在黑暗期(活跃期)开始后的前3 h (ZT12~15)出现觉醒时长减少、NREM和REM睡眠时长增加(
P
<
0.05)。另外
在该时期
敲除MT
3
R/Nqo2基因后大鼠还出现睡眠/觉醒
转化次数显著增加
觉醒片段长度缩短
觉醒片段化现象(
P
<
0.05)。脑电能谱分析显示
与对照组大鼠相比
敲除组大鼠NREM睡眠期θ波能量升高(
P
<
0.05)。
结论
2
MT
3
R/Nqo2基因敲除使大鼠NREM睡眠期EEG高频能量异常升高
NREM睡眠期稳态恢复过程受损
导致大鼠在黑暗期(活跃期)开始后的前3 h (ZT12-15)觉醒减少
睡眠增加
觉醒片段化。
Abstract
Objective
2
To observe the sleep/wake behaviors and EEG power spectrum in melatonin type 3 receptor (MT
3
R/Nqo2) knockout rats with CRISPR/Cas9 gene editing tool in order to explore the receptor mechanism of melatonin in regulation of sleep/wake behaviors.
Methods
2
SD rats with MT
3
R/Nqo2 knockout were created by introducing a mutation in the MT
3
R/Nqo2 gene frame shift by nonhomologous recombination repair causing the gene dysfunction
and the knockout was confirmed at the genetically
transcriptionally
and morphologically by PCR
RT-PCR
and FISH
respectively.Then the changes in sleep/wake behaviors and in EEG power spectrum were observed in the knockout rats and control rats.
Results
2
After MT
3
R/Nqo2 knockout
the knockout rats showed decreased arousal time and increased NREM and REM sleep time during the first 3 h (ZT12~15) after the onset of the dark phase (active phase) when compared with the control rats (
P
<
0.05).In addition
a significant increase in the number of sleep/wake transitions
a reduction in the duration of wakefulness
and appearance of wake fragmentation were also observed after MT
3
R/Nqo2 knockout during this period (
P
<
0.05).EEG power spectrum analysis revealed that NREM sleep θ band energy was elevated in the knockout rats than the control rats (
P
<
0.05).
Conclusion
2
MT
3
R/Nqo2 knockout results in abnormal power spectrum of EEG especially high-frequency energy during NREM sleep
disrupts sleep homeostasi
s
leads to decreased arousal
increased sleep
and fragmented arousals in the first 3 h (ZT12~15) after the onset of the dark phase (active phase) in rats.
关键词
Keywords
references
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