Molecular, cellular and in vivo analyses of photoresponses and circadian rhythms in animals

动物光反应和昼夜节律的分子、细胞和体内分析

• 批准号: 14104003
• 负责人: FUKADA Yoshitaka
• 金额: $ 71.64万
• 依托单位: University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (S)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14104003/
• 关键词: Pineal gland; Suprachiasmatic nucleus; Photoreceptive molecule; Exorhodopsin; G-protein; MAPK; Peripheral clock; E4BP4; 概日時計; CREB; ゼブラフィッシュ; Per; グルコース; TIEG1; Bmal1; サーカディアンリズム; クリプトクローム; リン酸化; GSK-3β; 視細胞; ファルネシル化; 明順応; MAPキナーゼ

(1)Activation of Gil induced a phase-shift of the chick pineal circadian clock in a manner similar to that induced by light. In the promoter region of pinopsin gene, we identified a light-responsive element responsible for its light-dependent transcriptional activation.(2)Farnesylation of retinal rod G-protein transducin plays an important role in not only the cellular light-signaling but also the light-adaptation. We found that GRK7-1 expressed in the zebrafish retinal cones shows opsin-phosphorylating activity dozens-fold higher than that of rod-specific GRK1A.(3)Chick pineal and frog retinal MAPK was circadian-activated showing the peak activity at night. Within the mouse SCN, the profiles of the temporal and light activation of MAPK are both different between the dorsomedial and central regions of the SCN. We identified amino acid residues in BMAL1, CRY1 and CRY2 to be phosphorylated by MAPK and their phosphorylation-dependent functional changes. We also found that SCOP expressed abundantly in the SCN is a negative regulator of K-Ras and downstream MAPK. It was shown that p38 MAPK regulates the molecular clock in the daytime to have the phase-advancing effect.(4)When the chicks entrained to 12L : 12D conditions were exposed to prolonged light extending into the early night, pineal E4BP4 protein was kept at a high level, which delayed the morning induction of Per2 and delayed the phase of the pineal clock. E4BP4 was phosphorylated by CKIε, leading to its proteasomal degradation.(5)We found that glucose-stimulation of rat-1 cells resets the clock by a mechanism to which Tieg 1 and Vdup 1 contribute. TIEG 1 protein was in fact up-regulated upon glucose administration and it directly acts on the promoter of Bmall to inhibit the transcription.(6)Within the promoter region of pineal opsin exorhodopsin gene, we identified a cis-acting element, PIPE, that drives pineal-specific gene expression in the zebrafish.

（1）Gil的激活以类似于光诱导的方式诱导鸡松果体昼夜节律钟的相移。在pinopsin基因的启动子区，我们发现了一个光响应元件，负责其光依赖性转录激活。（2）视网膜视杆细胞G蛋白转导蛋白的法尼基化不仅在细胞光信号转导中起重要作用，而且在光适应中也起重要作用。我们发现在斑马鱼视网膜视锥细胞中表达的GRK 7 -1显示出比视杆细胞特异性GRK 1A高几十倍的视蛋白磷酸化活性。（3）鸡松果体和蛙视网膜MAPK活性呈昼夜节律性变化，夜间活性最高。在小鼠SCN内，MAPK的时间和光激活的轮廓在SCN的背内侧和中央区域之间都是不同的。我们鉴定了BMAL 1、BMAL 1和BMAL 2中被MAPK磷酸化的氨基酸残基及其磷酸化依赖的功能变化。我们还发现SCOP在SCN中大量表达，是K-Ras和下游MAPK的负调节因子。结果表明，p38 MAPK在白天调节分子钟，具有时相推进效应。（4）当小鸡在12 L：12 D条件下长时间光照至深夜时，松果体E4 BP 4蛋白保持在较高水平，从而延迟了早晨Per 2的诱导，并延迟了松果体时钟的相位。E4 BP 4被CKIε磷酸化，导致其蛋白酶体降解。(5)We发现葡萄糖刺激大鼠-1细胞通过Tieg 1和Vdup 1贡献的机制重置时钟。TIEG 1蛋白事实上在葡萄糖给药后上调，并且其直接作用于Bmall的启动子以抑制转录。（6）在斑马鱼松果体视蛋白exorhodopsin基因的启动子区，我们发现了一个顺式作用元件PIPE，它驱动松果体特异性基因的表达。

项目成果

Suprachiasmatic nucleus circadian oscillatory protein, a novel binding partner of K-Ras in the membrane rafts, negatively regulates MAPK pathway

• DOI: 10.1074/jbc.m213214200
• 发表时间: 2003-04-25
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Shimizu, K;Okada, M;Fukada, Y
• 通讯作者: Fukada, Y

シリーズ21世紀の動物科学 : 第9巻『動物の感覚とリズム』の序章 : 感覚系と生物時計の対比と連関の項を分担執筆

21世纪动物科学丛书：第9卷《动物感官与节律》前言：与人合着感觉系统与生物钟的对比与关系部分。

• 发表时间: 2007
• 作者: 深田 吉孝(分担執筆)

『生物科学ニュース』の「動物の光受容と生物時計の分子メカニズムに関する研究(平成18年度 日本動物学会賞 受賞研究内容)」の項を執筆

为《生物科学新闻》撰写了“动物光感受和生物钟的分子机制研究（2006年日本动物学会获奖研究内容）”部分。

• 发表时间: 2007
• 作者: 深田 吉孝

実験医学増刊号『ダイナミックに新展開する脂質研究』の「Gタンパク質の脂質修飾を介したシグナル伝達の調節」の項を分担執筆

在实验医学特刊《脂质研究动态新进展》中共同撰写“通过G蛋白脂质修饰调节信号转导”章节

• 发表时间: 2005
• 作者: 深田 吉孝(分担執筆)

『図説 分子病態学』改訂3版 の「視覚異常症」の項を分担執筆

共同撰写《分子病理学图解》第三修订版中“视觉异常”部分

• 发表时间: 2003
• 作者: Ogura T;Kobayashi H;Ueoka Y;Okugawa K;Kato K;Hirakawa T;Hashimoto S;Taniguchi S;Hashimoto S;Wake N;Nakano H;深田 吉孝(分担執筆)
• 通讯作者: 深田 吉孝(分担執筆)