Graduate School of Innovative Life Science, University of Toyama, 3190 Gofuku, Toyama 930-8555, Japan.
Nat Commun. 2012 Jun 19;3:909. doi: 10.1038/ncomms1922.
Rhythmic expression of period (per) and timeless (tim) genes in central circadian pacemaker neurons and prothoracic gland cells, part of the peripheral circadian oscillators in flies, may synergistically control eclosion rhythms, but their oscillatory profiles remain unclear. Here we show differences and interactions between peripheral and central oscillators using per-luciferase and cytosolic Ca(2+) reporter (yellow cameleon) imaging in organotypic prothoracic gland cultures with or without the associated central nervous system. Isolated prothoracic gland cells exhibit light-insensitive synchronous per-transcriptional rhythms. In prothoracic gland cells associated with the central nervous system, however, per transcription is markedly amplified following 12-h light exposure, resulting in the manifestation of day-night rhythms in nuclear PER immunostaining levels and spontaneous Ca(2+) spiking. Unlike PER expression, nuclear TIM expression is associated with day-night cycles that are independent of the central nervous system. These results demonstrate that photoreception and synaptic signal transduction in/from the central nervous system coordinate molecular 'gears' in endocrine oscillators to generate physiological rhythms.
节律性表达周期(per)和无时间(tim)基因在中央生物钟神经元和前胸腺细胞中,它们是果蝇外周生物钟振荡器的一部分,可能协同控制蜕皮节律,但它们的振荡模式仍不清楚。在这里,我们使用前胸腺组织培养物中的 per-荧光素酶和胞质 Ca(2+)报告(黄色 cameleon)成像,显示了外周和中央振荡器之间的差异和相互作用,这些培养物有无相关的中枢神经系统。分离的前胸腺细胞表现出对光不敏感的同步 per-转录节律。然而,与中枢神经系统相关的前胸腺细胞中,per 转录在 12 小时光照暴露后显著放大,导致核 PER 免疫染色水平和自发 Ca(2+)爆发出现昼夜节律。与 PER 表达不同,核 TIM 表达与昼夜周期相关,与中枢神经系统无关。这些结果表明,中枢神经系统内/外的光感受和突触信号转导协调内分泌振荡器中的分子“齿轮”以产生生理节律。