Paradoxical (REM) sleep deprivation in mice using the small-platforms-over-water method: polysomnographic analyses and melanin-concentrating hormone and hypocretin/orexin neuronal activation before, during and after deprivation
Paradoxical (REM) sleep deprivation in mice using the small-platforms-over-water method: polysomnographic analyses and melanin-concentrating hormone and hypocretin/orexin neuronal activation before, during and after deprivation
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DOI：
10.1111/jsr.12269
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发表时间：
2015-06-01
影响因子：
4.4
通讯作者：
Peyron, Christelle
中科院分区：
文献类型：
--
作者：
Arthaud, Sebastien;Varin, Christophe;Peyron, Christelle
研究矛盾睡眠稳态需要具体有效地剥夺矛盾睡眠并评估随后的恢复期。为了实现这一目标，水上小平台技术已在大鼠中广泛使用，但仅在小鼠中进行了极少数研究，并且没有报告剥夺期间的睡眠数据。随着转基因小鼠和光遗传学等技术的出现，小鼠的使用越来越多，这就增加了对操纵矛盾睡眠的可靠方法的需求。为了满足这一需求，我们改进了这种剥夺方法，并在整个协议期间彻底分析了警惕状态。我们还使用 Fos 免疫组织化学研究了下丘脑分泌素/食欲素和黑色素浓缩激素神经元的激活，以验证小鼠调节矛盾睡眠的机制是否与大鼠相似。我们发现，48 小时的剥夺是非常有效的，残余反常睡眠量仅为 2.2%。慢波睡眠和清醒时间与基线相似，但在剥夺的前 4 小时内，慢波睡眠大幅减少。剥夺睡眠后，我们观察到在反弹的第一个小时内矛盾睡眠量增加了 124%。此外，34% 的下丘脑分泌素/食欲素神经元在睡眠剥夺期间被激活，而黑色素浓缩激素神经元仅在反常睡眠反弹期间被激活。皮质酮水平在剥夺后增加了一倍，并在恢复 4 小时后恢复到基线水平。总之，使用小平台水上方法可以在小鼠中获得相当选择性的剥夺和显着的反常睡眠反弹。与大鼠一样，反弹伴随着黑色素浓缩激素神经元的选择性激活。
Studying paradoxical sleep homeostasis requires the specific and efficient deprivation of paradoxical sleep and the evaluation of the subsequent recovery period. With this aim, the small-platforms-over-water technique has been used extensively in rats, but only rare studies were conducted in mice, with no sleep data reported during deprivation. Mice are used increasingly with the emergence of transgenic mice and technologies such as optogenetics, raising the need for a reliable method to manipulate paradoxical sleep. To fulfil this need, we refined this deprivation method and analysed vigilance states thoroughly during the entire protocol. We also studied activation of hypocretin/orexin and melanin-concentrating hormone neurones using Fos immunohistochemistry to verify whether mechanisms regulating paradoxical sleep in mice are similar to those in rats. We showed that 48h of deprivation was highly efficient, with a residual amount of paradoxical sleep of only 2.2%. Slow wave sleep and wake quantities were similar to baseline, except during the first 4h of deprivation, where slow wave sleep was strongly reduced. After deprivation, we observed a 124% increase in paradoxical sleep quantities during the first hour of rebound. In addition, 34% of hypocretin/orexin neurones were activated during deprivation, whereas melanin-concentrated hormone neurones were activated only during paradoxical sleep rebound. Corticosterone level showed a twofold increase after deprivation and returned to baseline level after 4h of recovery. In summary, a fairly selective deprivation and a significant rebound of paradoxical sleep can be obtained in mice using the small-platforms-over-water method. As in rats, rebound is accompanied by a selective activation of melanin-concentrating hormone neurones.