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.
