一般口演（Oral）

Oral Sleep and stress 一般口演睡眠とストレス

7月28日（日）9:45～10:00　第8会場（朱鷺メッセ　3F　303+304） 4O-08m2-1 NREM sleep is modulated by a subset of zinc-activated neurons in mice Yoan Cherasse（Cherasse Yoan）¹，Olusolape Ladenika（Ladenika Olusolape）¹，Gabrielle Boix（Boix Gabrielle）¹，Yoshihiro Urade（Urade Yoshihiro）²，Takeshi Sakurai（Sakurai Takeshi）¹ ¹University of Tsukuba ²The University of Tokyo Hospital Zinc is an essential mineral that plays an important role in the body by acting as a cofactor for more than 300 enzymes and 1000 transcription factors. While zinc is naturally present in food, particularly in meat and oyster, inappropriate or insufficient feeding behavior put more than 25% of the world population at risk of zinc deficiency. The purpose of our study was to examine the effect of zinc on sleep. We examined the sleep-promoting activity of zinc by monitoring locomotor activity and electroencephalogram after oral administration to mice at the onset of dark time. Compared with the placebo group, zinc-containing yeast extract (80mg/kg zinc) as well as zinc gluconate (40mg/kg zinc) dose dependently reduced locomotor activity and increased the total amount of non-rapid eye movement (NREM) sleep in mice. To better understand the molecular mechanisms, we then performed a c-fos mapping 1 hour after zinc administration. We discovered a subset of neurons activated into a specific area of the extended amygdala. After expressing DREADD receptors specifically in these neurons, chemogenetic activation (DREADD hM3Dq) increased the amount of NREM sleep, while their chemogenetic inhibition (DREADD hM4Di) could partially block the zinc-induced sleep. These results demonstrate the ability for these neurons to modulate the amount of NREM sleep. We are now performing experiments to better comprehend the role and function of these neurons as well as their connections with other brain regions.		7月28日（日）10:00～10:15　第8会場（朱鷺メッセ　3F　303+304） 4O-08m2-2 睡眠制御回路のコネクトミクス Masayoshi Ito（伊藤正芳）¹，Kei Ito（伊藤啓）^1,2，Gelard M Rubin（Rubin M Gelard）¹ ¹HHMI Janelia Research Campus ²Inst for Zoology, Univ of Cologne, Cologne, Germany Hypothalamus is a major center of neuroendocrine system that is involved in the regulation of various functions such as circadian rhythm, feeding and courtship behavior. These functions are important for survival of not only human being but also the flies. The superior medial protocerebrum (SMP) is a brain region located in the most dorsal part of the Drosophila brain, and it receives projections of neurons that are known to be associated with circadian rhythm, feeding and courtship behavior. The SMP also have connections with the mushroom body (MB) and central complex (CX), which are the center of olfactory learning, motor control, and also sleep regulation. However, the whole aspect of neuronal circuits and functional relationship in the SMP is not well known. In Drosophila Melanogaster, the whole brain connectomes based on the electron microscopy is ongoing in Janelia research campus. The whole brain connectomes gives us the details of neuroanatomy and synaptic connections among clock cells, MB and CX neurons which related to the sleep regulation. However, not only whole brain connectomes, but also the functions of each neuron are important to understand the neural network in the brain. An excellent genetic tool called the split-Gal4 system enables us to induce gene expressions only in a very specific cell type. The DNA-binding domain (DBD) and transcription activation domain (AD) of the Gal4 protein, each expressed under the control of a different promoter, form functional GAL4 proteins to drive transgene expression only at the intersection of the expression patterns of the two promoters. We tried more than 12,000 combinations of AD and DBD lines and identified 250 types of cell specific split-Gal4 lines which are the upstream or downstream circuits of clock cells and MB and CX neurons which revealed by the EM connectomes. We performed activation screening with the combination of thermo-sensitive dTRPA1 and found that there might be specific time windows to control sleep or awaking circuits. These neurons are supposed to be the convergence interneurons among the known sleep regulating circuits in SMP.

7月28日（日）10:15～10:30　第8会場（朱鷺メッセ　3F　303+304） 4O-08m2-3 オレキシン受容体拮抗薬スボレキサントが呼吸に与える影響 Isato Fukushi（福士勇人）¹，Shigefumi Yokota（横田茂文）²，Kotaro Takeda（武田湖太郎）^1,3，Jiro Terada（寺田二郎）^1,4，Yasumasa Okada（岡田泰昌）¹ ¹国立病院機構村山医療セ・臨床研究セ ²島根大・医解剖・神経科学 ³藤田医科大・医療科学・リハビリ ⁴千葉大・呼吸内科 Suvorexant (Belsomra (R)), a dual blocker of the orexin receptors (OX1R and OX2R), inhibits the arousal system in the central nervous system, induces sleep, and has been widely used for treating insomnia. Because many of sleep-inducing drugs suppress ventilation, concerns could be raised whether suvorexant affect breathing. However, the effects of suvorexant on breathing have not been well clarified. To investigate this issue, we first conducted immunohistological analysis of the orexin receptor expression in the brainstem; expression of OX2R with markers of putative respiratory centers (CGRP/FoxP2, Phox2b and somatostatin) was analyzed in the mouse brainstem regions that are crucially important in respiratory control, i.e., in the parabrachial nucleus (PBN), parafacial respiratory group/retrotrapezoid nucleus (pFRG/RTN) and preBotzinger complex (preBotC), respectively. Then, we analyzed ventilatory parameters as well as EEG in room air, hypercapnic (5% CO₂) and hypoxic (10% O₂) conditions before and after administration of low and high doses (10 mg/kg and 90 mg/kg) suvorexant. Experiments were performed in unanesthetized adult male mice. Surgical procedures to implant EEG electrodes were carried out under anesthesia. The mice were allowed to recover from surgery for at least 1 week until recordings were conducted. Respiratory flow was non-invasively measured by whole body plethysmography. Respiratory parameters (tidal volume, respiratory rate, and minute ventilation) were calculated from respiratory flow. The EEG data were subjected to frequency analysis by spectrography to calculate the power, which is a hallmark of the sleep/waking state. The oxygen concentration in the chamber was monitored with an oxygen concentration analyzer using a polarographic sensor. In results, expression of OX2R was confirmed in Phox2b- and somatostatin-immunoreactive neurons in the pFRG/RTN and preBotC, respectively, suggesting the involvement of orexin in respiratory control. However, either dose of suvorexant did not affect ventilation in room air or hypoxic/hypercapnic ventilatory responses. We conclude that suvorexant could be safely used without suppressing breathing.		7月28日（日）10:30～10:45　第8会場（朱鷺メッセ　3F　303+304） 4O-08m2-4 環境ストレスによる脳内インスリンニューロンの低温応答の変容 Yusuke Hara（原佑介）¹，Noriyuki Ojima（小島紀幸）²，Hiroki Ito（伊藤弘樹）²，Daisuke Yamamoto（山元大輔）¹ ¹情報通信研未来ICT行動神経 ²東北大院生命科学 Organisms are inevitably exposed to various environmental stresses. Particularly, low temperature and scarcity of food in winter are deadly to most wild animals. To cope with such stresses, insects have developed an adaptive strategy of dormancy in which they minimize energy consumption and increase stress resistance to overcome unfavorable environmental conditions. In Drosophila melanogaster, adult females enter reproductive dormancy when they are exposed to the combination of low temperature, starvation, and short-day photoperiod. We have shown that brain insulin-producing cells (IPCs) regulate ovarian development and have a role in decision making for entering reproductive dormancy (Ojima et al., 2018). As in mammalian pancreatic β cells, nutritional states impact production and release of insulin-like peptides in IPCs, yet it remains unknown how IPCs integrate inputs encoding environmental states. Here we performed in vivo whole-cell patch clamp recordings from IPCs while changing ambient temperature. Under normal conditions (12L/12D, 25°C, fed for 1 wk), an acute exposure to low temperature markedly depolarized the IPC membrane, leading to spike firing. This response was mediated by Gustatory receptor 28b and a K2P channel, dTASK-7. In contrast, under dormancy-inducing conditions (10L/14D, 11°C, starved for 1 wk), IPCs barely generated spikes in response to a similar cold exposure. IPCs in newly emerged flies neither showed large depolarization nor firing during an acute cold challenge. These observations suggest that IPCs acquire the ability to generate a large depolarizing potential in response to an acute cold stimulus only after being exposed to environmental conditions that stimulate breeding (e.g., 12L/12D, 25°C, fed for 1 wk). Thus, the present study unravels a cellular mechanism underlying acclimatization.