The metabolism of phospholipids oscillates rhythmically in cultures of fibroblasts and is regulated by the clock protein PERIOD 1
The metabolism of phospholipids oscillates rhythmically in cultures of fibroblasts and is regulated by the clock protein PERIOD 1
复制标题
DOI：
10.1096/fj.03-0417fje
复制
发表时间：
2004-01-01
期刊：
影响因子：
4.8
通讯作者：
Guido, ME
中科院分区：
文献类型：
--
作者：
Marquez, S;Crespo, P;Guido, ME
哺乳动物的昼夜节律计时系统是由分布在全身的无数细胞振荡器和调节时间生理和行为的中枢起搏器组成的。在体内和培养物中，外周时钟显示基因表达的昼夜节律。我们研究了磷脂的生物合成以及时钟基因周期1（Per1）的表达，并在培养的静止NIH 3T3细胞同步的2小时血清休克的磷脂代谢的调节，其潜在的参与。在血清处理后0.5至62 h的阶段给予放射性标记前体30 min脉冲。我们观察到磷脂标记的每日节律至少持续两个周期，治疗后29和58小时水平显著降低。Per1的表达表现出快速和短暂的诱导和每日节律性的反相脂质标记。在Per1表达敲低后，磷脂标记的节律丧失。此外，在培养的时钟突变成纤维细胞与时钟机制的损伤，PERI是平等地表达在所有时间检查和磷脂标记没有振荡。结果表明，磷脂的生物合成振荡每天在培养的成纤维细胞的内源性时钟机制，涉及Per1的表达。
The mammalian circadian timing system is composed of countless cell oscillators distributed throughout the body and central pacemakers regulating temporal physiology and behavior. Peripheral clocks display circadian rhythms in gene expression both in vivo and in culture. We examined the biosynthesis of phospholipids as well as the expression of the clock gene period 1 (Per1) and its potential involvement in the regulation of the phospholipid metabolism in cultured quiescent NIH 3T3 cells synchronized by a 2 h serum shock. A 30 min pulse of radiolabeled precursor was given at phases ranging from 0.5 to 62 h after serum treatment. We observed a daily rhythm in the phospholipid labeling that persisted at least for two cycles, with levels significantly decreasing 29 and 58 h after treatment. Per1 expression exhibited a rapid and transient induction and a daily rhythmicity in antiphase to the lipid labeling. After Per1 expression knockdown, the rhythm of phospholipid labeling was lost. Furthermore, in cultures of CLOCK mutant fibroblasts-cells with a clock mechanism impairment-PERI was equally expressed at all times examined and the phospholipid labeling did not oscillate. The results demonstrate that the biosynthesis of phospholipids oscillates daily in cultured fibroblasts by an endogenous clock mechanism involving Per1 expression.