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为探讨中枢5羟色胺(5-HT)的缺失对正常睡眠和快速眼动睡眠剥夺(REM sleep deprivation)情况下orexin阳性神经元活动的影响,本研究利用中枢5-HT神经元缺失的条件性基因敲除小鼠(Pet1-Cre/Lmx1b flox/flox CKO小鼠),采用小平台水环境法建立小鼠快速眼动睡眠剥夺模型,免疫组化方法观察野生型小鼠和中枢5-HT神经元缺失小鼠在正常睡眠状态及8 h快速眼动睡眠剥夺后下丘脑内orexin阳性神经元的数量,免疫组化双标法观察orexin/c-fos双标神经元占orexin阳性神经元的比例。结果显示:CKO小鼠睡眠剥夺前后orexin阳性神经元的数量未见明显差别,与野生型小鼠相比亦未见统计学差别;在正常睡眠状态下(对照组),CKO小鼠orexin/c-fos双标神经元的数量与野生型小鼠相当,但睡眠剥夺后明显低于野生型小鼠睡眠剥夺组。本研究结果提示,作为维持觉醒的重要神经递质5-HT的缺失可能降低了中枢神经系统的觉醒水平,致使睡眠剥夺不能提高促发和维持觉醒的orexin阳性神经元的活性。
To investigate the effect of deletion of 5-hydroxytryptamine (5-HT) on the activity of orexin-positive neurons in normal sleep and REM sleep deprivation, we used the conditional gene deletion of central 5-HT neurons (Pet1-Cre / Lmx1b flox / flox CKO mice) were knocked out and the mouse model of rapid eye movement sleep deprivation was established by using the small platform water environment method. The expression of wild type mouse and central 5-HT neurons were observed by immunohistochemistry The number of orexin-positive neurons in the hypothalamus of normal mice and mice after 8-h REM sleep deprivation were observed. The ratio of orexin / c-fos double-labeled neurons to orexin-positive neurons was observed by immunohistochemistry. The results showed that there was no significant difference in the number of orexin positive neurons before and after sleep deprivation in CKO mice, and there was no significant difference between wild type mice and CKO mice. Under normal sleep conditions (control group), orexin / c The number of -fos double-labeled neurons was comparable to that of wild-type mice but significantly lower than that of wild-type mice after sleep deprivation. Our results suggest that the absence of 5-HT, an important neurotransmitter that maintains wakefulness, may reduce central nervous system arousal, rendering sleep deprivation incapable of increasing the activity of orexin-positive neurons that trigger and sustain arousal.