The rat ponto-medullary network responsible for paradoxical sleep onset and maintenance: a combined microinjection and functional neuroanatomical study
The rat ponto-medullary network responsible for paradoxical sleep onset and maintenance: a combined microinjection and functional neuroanatomical study
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DOI：
10.1046/j.1460-9568.2002.02257.x
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发表时间：
2002-11-01
影响因子：
3.4
通讯作者：
Luppi, PH
中科院分区：
文献类型：
--
作者：
Boissard, R;Gervasoni, D;Luppi, PH
与猫不同的是，之前还没有在老鼠身上发现负责矛盾睡眠（PS）发生和维持的神经网络。为了填补这一空白，本研究开发了一种新技术，包括记录未麻醉的头部受限大鼠的睡眠-觉醒状态，同时通过玻璃多管微离子导入微离子导入局部药物，进入脑桥背盖，并将其与功能神经解剖学相结合。离子电泳的药理药物包括卡巴醇、kainic酸、双柳碱和gabazine。然后通过相邻的微管装配桶注射顺行（phaseolus vulgaris白细胞凝集素）或逆行（霍乱毒素B亚基）示踪剂并通过C-Fos免疫染色来鉴定注射部位及其输出。双库克林、gabazine和kainic酸特异性地射入脑桥嗅觉下核（SLD），在几分钟内诱导出一种ps样状态，其特征是持续的肌肉弛缓、低电压脑电图和对刺激缺乏反应。相反，向SLD中喷射乙醇会引起清醒。在PHA-L、甘氨酸和C-Fos多重双标记实验中，可以看到来自SLD的顺行标记纤维与来自腹侧巨细胞和细胞旁网状核的甘氨酸和C-Fos阳性神经元（在药物诱导的ps样状态90分钟后标记）相对应。总之，这些数据表明，SLD核中包含的神经元群在PS的发生和维持中起着至关重要的作用。此外，他们认为这些神经元的gaba能去抑制和谷氨酸兴奋也可能在PS的发病中起关键作用。
The neuronal network responsible for paradoxical sleep (PS) onset and maintenance has not previously been identified in the rat, unlike the cat. To fill this gap, this study has developed a new technique involving the recording of sleep-wake states in unanaesthetized head-restrained rats whilst locally administering pharmacological agents by microiontophoresis from glass multibarrel micropipettes, into the dorsal pontine tegmentum and combining this with functional neuroanatomy. Pharmacological agents used for iontophoretic administration included carbachol, kainic acid, bicuculline and gabazine. The injection sites and their efferents were then identified by injections of anterograde (phaseolus vulgaris leucoagglutinin) or retrograde (cholera toxin B subunit) tracers through an adjacent barrel of the micropipette assembly and by C-Fos immunostaining. Bicuculline, gabazine and kainic acid ejections specifically into the pontine sublaterodorsal nucleus (SLD) induced within a few minutes a PS-like state characterized by a continuous muscle atonia, low voltage EEG and a lack of reaction to stimuli. In contrast, carbachol ejections into the SLD induced wakefulness. In PHA-L, glycine and C-Fos multiple double-labelling experiments, anterogradely labelled fibres originating from the SLD were seen apposed on glycine and C-Fos positive neurons (labelled after 90 min of pharmacologically induced PS-like state) from the ventral gigantocellular and parvicellular reticular nuclei. Altogether, these data indicate that the SLD nuclei contain a population of neurons playing a crucial role in PS onset and maintenance. Furthermore, they suggest that GABAergic disinhibition and glutamate excitation of these neurons might also play a crucial role in the onset of PS.