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Central and peripheral clocks; Mechanisms of temporal integration

中央和外围时钟；

基本信息

批准号：
17390059
负责人：
HONMA Sato
金额：
$ 9.41万
依托单位：
Hokkaido University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2005
资助国家：
日本
起止时间：
2005 至 2006
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390059/
关键词：
Physiology Neuroscience Signal transduction Biological Clock Bioluminescence

项目摘要

We examined the mechanisms how the multiple oscillator cells in the suprashicasmatic nucleus (SCN) synchronize to each other and entrain to the external light cue to orchestrate peripheral clocks. We also studied the synchronization mechanisms of the peripheral clocks by the central clock in the SCN. We established two types of luciferase reporter transgenic mice for this study.1.Establishment of transgenic mice: We established a transgenic mouse expressing firefly luciferase under the control of Per1 promoter (Per1-luc) and a dual reporter transgenic mouse bearing secreting luciferase (VL) under Per1 promoter and non-secreting luciferase (FL) under Bmal1 promoter (PVB1). Behavioral rhythms and responses to lights were not different in these transgenic mice from wild type controls.2.Multioscillatory structure in the central clock: Per1-luc mice entrained to LD12:12 were exposed to either LD18:6 or LD6:18. After monitoring behavioral rhythms, we made two serial coronal slices and monitored bioluminescence for 5 consecutive days. Robust Pen expression rhythms were detected with a peak around the subjective noon in all SCN slices examined but anterior SCN under LD18:6 in which bimodal Pen peaks were detected. Irrespective of photoperiods, Pen peaks in the anterior SCN correlated with activity onset, while that in the posterior SCN, with the activity end, suggesting the localization of E and M oscillator in the anterior and posterior SCN, respectively.3.Autonomous oscillation and phase-adjustment of the peripheral clocks: Various peripheral tissues of PVBF mice and Bmal1 expression rhythms were measured. In all peripheral tissues examined, robust Bmal1 rhythms were detected with the periods as well as the peak phases were specific to tissues. Long-term culture of liver slices showed autonomous rhythms continuation and phase-dependent phase set by medium exchange, suggesting coupling of single cell oscillators.

我们研究了下丘脑上核（SCN）中的多个振荡细胞如何相互同步并与外部光线索一起协调外周时钟的机制。研究了SCN中中心时钟对外围时钟的同步机制。本研究建立了两种荧光素酶报告基因转基因小鼠。1.转基因小鼠的建立：建立了Per 1启动子调控下的萤火虫荧光素酶转基因小鼠（Per 1-luc）和Per 1启动子调控下的分泌型荧光素酶（VL）和Bmal 1启动子调控下的非分泌型荧光素酶（FL）双报告基因转基因小鼠（PVB 1）。行为节律和对光的反应在这些转基因小鼠中与野生型对照没有差异。2.中央时钟的多振荡结构：将携带LD 12：12的Per 1-luc小鼠暴露于LD 18：6或LD 6：18。在监测行为节律后，我们制作了两个连续的冠状切片，并连续5天监测生物发光。在所有检查的SCN切片中检测到稳健的Pen表达节律，其中峰在主观中午附近，但在LD 18：6下的前SCN中检测到双峰Pen峰。不考虑光周期，前SCN的Pen峰与活动开始相关，后SCN的Pen峰与活动结束相关，表明E振荡器和M振荡器分别位于前SCN和后SCN。3.外周时钟的自主振荡和相位调节：检测PVBF小鼠不同外周组织和Bmal 1表达节律。在检查的所有外周组织中，检测到稳健的Bmal 1节律，其周期以及峰值相位是特定于组织的。长期培养的肝切片显示自主节律的持续和相依赖的阶段设置的介质交换，这表明耦合的单细胞振荡器。