Circadian profiling of the transcriptome in NIH/3T3 fibroblasts: comparison with rhythmic gene expression in SCN2.2 cells and the rat SCN
Circadian profiling of the transcriptome in NIH/3T3 fibroblasts: comparison with rhythmic gene expression in SCN2.2 cells and the rat SCN
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DOI：
10.1152/physiolgenomics.00199.2006
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发表时间：
2007-05-11
影响因子：
4.6
通讯作者：
Earnest, David J.
中科院分区：
文献类型：
--
作者：
Menger, Gus J.;Allen, Gregg C.;Earnest, David J.
为了筛选可以区分哺乳动物视交叉上核（SCN）中的起搏器与外周型振荡器（其中典型的时钟装置以昼夜节律的方式类似地调节）的输出信号，分析了毛喉素刺激的NIH/3 T3成纤维细胞中转录组的节律行为，并相对于体外SCN 2.2细胞和大鼠SCN进行了比较。与SCN 2.2和大鼠SCN转录组的昼夜节律分析相似，NIH/3 T3成纤维细胞在核心时钟基因Per 2、Cry 1和Bmal 1以及323种功能多样的转录物的表达中表现出昼夜节律波动，其中许多转录物调节细胞通讯。NIH/3 T3成纤维细胞、SCN 2.2细胞和大鼠SCN之间节律性转录物的重叠仅限于这些时钟基因和其他四个介导脂肪酸和脂质代谢或作为核因子发挥功能的基因。与NIH/3 T3细胞相比，SCN振荡器中调节葡萄糖代谢和神经传递的基因的昼夜节律表达更为普遍。再加上SCN 2.2细胞和大鼠SCN中诱导型一氧化氮合酶（iNos）的节律调节证据，而不是在成纤维细胞中，研究NOS抑制剂对SCN 2.2细胞和未处理的NIH/3 T3细胞共培养物中代谢节律的影响表明，气态神经递质一氧化氮可能在SCN起搏功能中发挥关键作用。SCN和非SCN细胞中的昼夜节律基因表达的这种比较分析可能在选择性分析参与SCN振荡器的耦合和下游细胞中的节律性调节的昼夜节律信号中具有重要意义。
To screen for output signals that may distinguish the pacemaker in the mammalian suprachiasmatic nucleus (SCN) from peripheral-type oscillators in which the canonical clockworks are similarly regulated in a circadian manner, the rhythmic behavior of the transcriptome in forskolin-stimulated NIH/3T3 fibroblasts was analyzed and compared relative to SCN2.2 cells in vitro and the rat SCN. Similar to the circadian profiling of the SCN2.2 and rat SCN transcriptomes, NIH/3T3 fibroblasts exhibited circadian fluctuations in the expression of the core clock genes, Per2, Cry1, and Bmal1, and 323 functionally diverse transcripts, many of which regulate cellular communication. Overlap in rhythmic transcripts among NIH/3T3 fibroblasts, SCN2.2 cells, and the rat SCN was limited to these clock genes and four other genes that mediate fatty acid and lipid metabolism or function as nuclear factors. Compared with NIH/3T3 cells, circadian gene expression in SCN oscillators was more prevalent among genes mediating glucose metabolism and neurotransmission. Coupled with evidence for the rhythmic regulation of the inducible isoform of nitric oxide synthase (iNos) in SCN2.2 cells and the rat SCN but not in fibroblasts, studies examining the effects of a NOS inhibitor on metabolic rhythms in cocultures containing SCN2.2 cells and untreated NIH/3T3 cells suggest that the gaseous neurotransmitter nitric oxide may play a key role in SCN pacemaker function. This comparative analysis of circadian gene expression in SCN and non-SCN cells may have important implications in the selective analysis of circadian signals involved in the coupling of SCN oscillators and regulation of rhythmicity in downstream cells.