Establishment of an in vitro cell line experimental system for the studablishment of an in vitro cell line experimental system for the study oy of inhalational anesthetic mechanisms.
Establishment of an in vitro cell line experimental system for the studablishment of an in vitro cell line experimental system for the study oy of inhalational anesthetic mechanisms.
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体外细胞系实验体系的建立用于建立用于吸入麻醉机制研究的体外细胞系实验体系。
DOI：
10.1016/j.neulet.2016.04.005
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Ozawa H.
中科院分区：
文献类型：
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作者：
Nagamoto S;Iijima N;Ishii H;Takumi K;Higo S;Aikawa S;Anzai M;Matsuo I;Nakagawa S;Takashima N;Shigeyoshi Y;Sakamoto A;Ozawa H.
全身麻醉影响各器官中时钟基因的表达。Per 2（生物钟的核心成分）的表达在视交叉上核（SCN）中受到七氟烷显著且可逆的抑制，被认为是大脑中麻醉作用的生化标志物。SCN包含各种类型的神经元，这种复杂性使得研究麻醉的分子机制变得困难。在这里，我们使用细胞系建立了一个体外实验系统来研究麻醉作用的机制。该系统的开发包括两个步骤：首先，我们开发了一个系统的应用吸入麻醉剂和孵育;接下来，我们建立了麻醉反应细胞表达mPer 2 promoter-dLuc的培养物。源自小鼠下丘脑的GT 1 -7细胞通过可逆地降低mPer 2启动子驱动的生物发光来对七氟烷做出反应。有趣的是，仅在血清饥饿的GT 1 -7细胞中发现生物发光的抑制，其显示出神经元样形态，但在生长的细胞中没有发现，这表明神经元样特征是GT 1 -7细胞中的麻醉作用所需的。
General anesthesia affects the expression of clock genes in various organs. Expression ofPer2, a core component of the circadian clock, is markedly and reversibly suppressed by sevoflurane in the suprachiasmatic nucleus (SCN), and is considered to be a biochemical marker of anesthetic effect in the brain. The SCN contains various types of neurons, and this complexity makes it difficult to investigate the molecular mechanisms of anesthesia. Here, we established anin vitroexperimental system using a cell line to investigate the mechanisms underlying anesthetic action. Development of the system comprised two steps: first, we developed a system for application of inhalational anesthetics and incubation; next, we established cultures of anesthetic-responsive cells expressingmPer2promoter-dLuc. GT1-7 cells, derived from the mouse hypothalamus, responded to sevoflurane by reversibly decreasingmPer2-promoter-driven bioluminescence. Interestingly, the suppression of bioluminescence was found only in the serum-starved GT1-7 cells, which showed neuron-like morphology, but not in growing cells, suggesting that neuron-like characteristics are required for anesthetic effects in GT1-7 cells.