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THE MECHANISM OF EXPRESSION OF CIRCADIAN CLOCK RELATED PROTEINS IN AN INSECT OPTIC LOBE

昆虫视神经节律相关蛋白的表达机制

基本信息

批准号：
10640667
负责人：
TOMIOKA Kenji
金额：
$ 2.3万
依托单位：
YAMAGUCHI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10640667/
关键词：
cricket optic lobe protein synthesis circadian clock PERIOD 昆虫蛋白合成時計蛋白質転写概日リズム

项目摘要

We investigated the mechanism of expression of clock related proteins in the optic lobe of the cricket Gryllus bimaculatus. 1) The effects of a translation inhibitor, cycloheximide (CHX), on the circadian neuronal activity rhythm of the optic lamina-medulla compound eye complex cultured in vitro were examined. Continuous application of CHX abolished circadian rhythms of both the spontaneous neuronal activity and the visually evoked response. However, it abolished neither the spontaneous activity nor the visually evoked response. 2) When the complex was treated with 10^<-5> M CHX for 6 h, the rhythm exhibited a marked phase shift. The magnitude and direction of the phase shift were dependent on the phase at which the complex was treated with CHX ; phase delays occurred during the late subjective day to early subjective night, whereas phase advances occurred around the late subjective night. 3) The phase response curve for a reversible transcription inhibitor, 5,6-dichlorobenzimidazole riboside (DRB), shows both phase delay and advance 6hrs earlier than that for CHX.4) The abundance of 31kDa protein, which shows a circadian rhythm in its abundance, was reduced when the rhythm was shifted by CHX or DRB.5) PDF and PERIOD, which are involved in the circadian rhythm in Drosophila, also showed circadian rhythmic change in their abundance in the optic lobe. 6) Immunohistochemistry revealed that PERIOD protein is expressed in cells located in the outer chiasma between the lamina and the medulla. These results suggest that gene expression including transcription and translation is a required process in the cricket optic lobe circadian clock, and that PDF and PERIOD may be involved in the clock mechanism.

我们研究了时钟相关蛋白在蟋蟀视叶中的表达机制。1)观察了翻译抑制剂放线菌酮(CHX)对体外培养的视板-延髓复合体昼夜节律的影响。持续应用CHX可使自发神经元活动和视觉诱发反应的昼夜节律消失。然而，它既没有消除自发活动，也没有消除视觉诱发的反应。2)10^-lt；-5-mCHX作用6h后，节律发生明显的相移。相移的大小和方向取决于用CHX治疗复合体的相位；相位延迟发生在主观白天晚期到主观夜晚早些时候，而相位推进发生在主观夜间深夜。3)可逆转录抑制剂5，6-二氯苯并咪唑核苷(DRB)的时相反应曲线比CHX提前6小时。4)CHX或DRB改变节律后，31 kDa蛋白的丰度降低。5)参与果蝇昼夜节律的PDF和PERIOD在视叶中的丰度也表现出昼夜节律的变化。6)免疫组织化学结果显示，Period蛋白表达于板层与延髓之间的外交叉细胞。这些结果表明，包括转录和翻译在内的基因表达是蟋蟀视叶昼夜节律钟所必需的过程，而PDF和Period可能参与了时钟机制。