title: Establishment of an in vitro cell line experimental system for the study of inhalational anesthetic mechanisms.
abstract: 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.
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan; Department of Anesthesiology and Pain Medicine, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Seiji
authors.initials: S
authors.lastname: Nagamoto
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan. Electronic address: niijima@nms.ac.jp.
authors.forename: Norio
authors.initials: N
authors.lastname: Iijima
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Hirotaka
authors.initials: H
authors.lastname: Ishii
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Takumi
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Shimpei
authors.initials: S
authors.lastname: Higo
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Satoko
authors.initials: S
authors.lastname: Aikawa
authors.affiliation: Department of Anesthesiology and Pain Medicine, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Megumi
authors.initials: M
authors.lastname: Anzai
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan; Department of Anesthesiology and Pain Medicine, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Izumi
authors.initials: I
authors.lastname: Matsuo
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan; Department of Anesthesiology and Pain Medicine, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Nakagawa
authors.affiliation: Department of Anatomy and Neurobiology, Kinki University Faculty of Medicine, Osakasayama, Osaka 589-8511, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Takashima
authors.affiliation: Department of Anatomy and Neurobiology, Kinki University Faculty of Medicine, Osakasayama, Osaka 589-8511, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Shigeyoshi
authors.affiliation: Department of Anesthesiology and Pain Medicine, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Atsuhiro
authors.initials: A
authors.lastname: Sakamoto
authors.affiliation: Department of Anatomy and Neurobiology, Graduate School of Medicine, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Ozawa
mesh_terms: D000777:Anesthetics / Q000494:pharmacology*; D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line / Q000166:cytology* / Q000187:drug effects*; D002940:Circadian Rhythm; D008163:Luminescent Measurements; D008738:Methyl Ethers / Q000494:pharmacology*; D051379:Mice; D051381:Rats; D000077149:Sevoflurane; D019076:Transgenes
chemical_list: D000777:Anesthetics; D018685:Anesthetics, Inhalation; D008738:Methyl Ethers; D000077149:Sevoflurane
keywords: Circadian rhythm; GT1-7; Inhalational anesthetic; Per2; Sevoflurane