Establishment of an in vitro cell line experimental system for the study of inhalational anesthetic mechanisms

Seiji Nagamoto
Norio Iijima
Hirotaka Ishii
Ken Takumi
Shimpei Higo
Satoko Aikawa
Megumi Anzai
Izumi Matsuo
Shinji Nakagawa
Naoyuki Takashima
Yasufumi Shigeyoshi
Atsuhiro Sakamoto
Hitoshi Ozawa

General anesthesia affects the expression of clock genes in various organs. Expression of Per2
 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 an in vitro experimental 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 expressing mPer2 promoter-dLuc. GT1-7 cells
 derived from the mouse hypothalamus
 responded to sevoflurane by reversibly decreasing mPer2-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. (C) 2016 Elsevier Ireland Ltd. All rights reserved.

NEUROSCIENCE LETTERS

ELSEVIER IRELAND LTD

620

10.1016/j.neulet.2016.04.005

https://doi.org/10.1016/j.neulet.2016.04.005
https://www.ncbi.nlm.nih.gov/pubmed/27057734
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