de Carvalho Magalhães Moraes Maria Nathália, de Oliveira Poletini Maristela, Ramos Bruno Cesar Ribeiro, de Lima Leonardo Henrique Ribeiro Graciani, de Lauro Castrucci Ana Maria
Department of Physiology, Institute of Biosciences, University of São Paulo, São Paulo, Brazil.
Photochem Photobiol. 2013 Dec 26. doi: 10.1111/j.1751-1097.2013.12230.x.
Light-dark cycles are considered important cues to entrain biological clocks. A feedback loop of clock gene transcription and translation is the molecular basis underlying the mechanism of both central and peripheral clocks. Xenopus laevis embryonic melanophores respond to light with melanin granule dispersion, response possibly mediated by the photopigment melanopsin. In order to test whether light modulates clock gene expression in Xenopus melanophores, we used qPCR to evaluate the relative mRNA levels of Per1, Per2, Clock and Bmal1 in cultured melanophores exposed to light-dark (LD) cycle or constant darkness (DD). LD cycles elicited temporal changes in the expression of Per1, Per2 and Bmal1. A 10-min pulse of blue light was able to increase the expression of Per1 and Per2. Red light had no effect on the expression of these clock genes. These data suggest the participation of a blue-wavelength sensitive pigment in the light-dark cycle-mediated oscillation of the endogenous clock. Our results add an important contribution to the emerging field of peripheral clocks, which in non-mammalian vertebrates have been mostly studied in Drosophila and Danio rerio. Within this context, we show that Xenopus laevis melanophores, which have already led to melanopsin discovery, represent an ideal model to understanding circadian rhythms. This article is protected by copyright. All rights reserved.
明暗周期被认为是调节生物钟的重要线索。生物钟基因转录和翻译的反馈环是中枢和外周生物钟机制的分子基础。非洲爪蟾胚胎黑素细胞通过黑色素颗粒分散对光作出反应,这种反应可能由光色素黑素视蛋白介导。为了测试光是否调节非洲爪蟾黑素细胞中的生物钟基因表达,我们使用定量聚合酶链反应(qPCR)来评估暴露于明暗(LD)周期或持续黑暗(DD)的培养黑素细胞中Per1、Per2、Clock和Bmal1的相对mRNA水平。LD周期引起Per1、Per2和Bmal1表达的时间变化。10分钟的蓝光脉冲能够增加Per1和Per2的表达。红光对这些生物钟基因的表达没有影响。这些数据表明,一种对蓝光波长敏感的色素参与了明暗周期介导的内源性生物钟振荡。我们的结果为外周生物钟这一新兴领域做出了重要贡献,在非哺乳动物脊椎动物中,外周生物钟主要在果蝇和斑马鱼中进行了研究。在此背景下,我们表明,已经导致黑素视蛋白发现的非洲爪蟾黑素细胞是理解昼夜节律的理想模型。本文受版权保护。保留所有权利。