Effects of knockout of MT<sub>3</sub>R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats

Xin LI; Shiyin LI; Fenlan LUO; Shuancheng REN; Zhi'an HU; Chao HE

doi:10.16016/j.2097-0927.202212125

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Effects of knockout of MT₃R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats

Neuroscience | 更新时间：2026-01-09

- Effects of knockout of MT₃R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats
- Journal of Army Medical University Vol. 45, Issue 9, Pages: 936-946(2023)
- 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：
- 基金信息：
  
  the National Natural Science Foundation of China(31970945);Chongqing Science Funds for Distinguished Young Scholars(CSTB2022NSCQ-JQX0019);Science and Technology Innovation 2030 - Major Program from the Ministry of Science and Technology of China(2021ZD0203200-01)
- DOI：10.16016/j.2097-0927.202212125
  CLC： R-332;R338.63;R338.8
- Received：20 December 2022，
  
  Revised：2023-02-03，
  
  Published：15 May 2023
- 稿件说明：
移动端阅览
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：

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

利用CRISPR/Cas9基因编辑工具构建褪黑素3型受体(MT

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

利用非同源重组修复引入突变的方式

造成MT

R/Nqo2基因蛋白读码框移码突变

使该基因功能缺失

建立MT

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

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(

0.05)。另外

在该时期

敲除MT

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(

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(

0.05)。

结论

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

To observe the sleep/wake behaviors and EEG power spectrum in melatonin type 3 receptor (MT

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

SD rats with MT

R/Nqo2 knockout were created by introducing a mutation in the MT

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

After MT

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 (

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

R/Nqo2 knockout during this period (

0.05).EEG power spectrum analysis revealed that NREM sleep θ band energy was elevated in the knockout rats than the control rats (

0.05).

Conclusion

R/Nqo2 knockout results in abnormal power spectrum of EEG especially high-frequency energy during NREM sleep

disrupts sleep homeostasi

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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Effects of knockout of MT3R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats

DOI：10.16016/j.2097-0927.202212125

摘要

Abstract

关键词

Keywords

references

Effects of knockout of MT₃R/Nqo2 gene on sleep/wake behaviors and EEG power spectrum in rats