Fibroblast circadian rhythms of PER2 expression depend on membrane potential and intracellular calcium.
Fibroblast circadian rhythms of PER2 expression depend on membrane potential and intracellular calcium.
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DOI：
10.3109/07420528.2012.679330
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发表时间：
2012-07
影响因子：
2.8
通讯作者：
Welsh DK
中科院分区：
文献类型：
--
作者：
Noguchi T;Wang CW;Pan H;Welsh DK
下丘脑的视交叉上核（SCN）同步全身细胞和组织的昼夜节律。在SCN神经元中，时钟基因表达的节律被操纵抑制，使质膜超极化或降低细胞内Ca2+。然而，其他细胞中的时钟是否也依赖于膜电位和钙是未知的。在这项研究中，我们研究了膜电位和细胞内钙对小鼠原代成纤维细胞昼夜节律的影响。使用PER2::LUC敲入报告基因监测时钟基因表达的节律。我们发现在较低（超极化）K+浓度下，节律丢失或延迟。生物发光成像显示，培养中节律性的丧失是由于单个细胞的节律性丧失，而不是细胞之间的不同步。在Ca2+浓度较低的情况下，节律提前或周期缩短。通过钙螯合剂1,2-双（2-氨基苯氧基）乙烷-N，N,N ',N ' -四乙酸四乙酰氧基甲酯（BAPTA-AM）缓冲细胞内Ca2+或通过thapsigargin延迟节律操纵ip3敏感的细胞内钙储存。这些结果表明，成纤维细胞的生物钟，如在SCN神经元中，是由膜电位和Ca2+调节的。我们观察到细胞内Ca2+的变化可能介导膜电位的影响。
The suprachiasmatic nucleus (SCN) of the hypothalamus synchronizes circadian rhythms of cells and tissues throughout the body. In SCN neurons, rhythms of clock gene expression are suppressed by manipulations that hyperpolarize the plasma membrane or lower intracellular Ca2+. However, whether clocks in other cells also depend on membrane potential and calcium is unknown. In this study, we investigate the effects of membrane potential and intracellular calcium on circadian rhythms in mouse primary fibroblasts. Rhythms of clock gene expression were monitored using a PER2::LUC knockin reporter. We found that rhythms were lost or delayed at lower (hyperpolarizing) K+ concentrations. Bioluminescence imaging revealed that this loss of rhythmicity in cultures was due to loss of rhythmicity of single cells rather than desynchrony among cells. In lower Ca2+ concentrations, rhythms were advanced or had shorter periods. Buffering intracellular Ca2+ by the calcium chelator 1,2-Bis(2-aminophenoxy) ethane-N,N,N′,N′-tetraacetic acid tetrakis acetoxymethyl ester (BAPTA-AM) or manipulation of IP3-sensitive intracellular calcium stores by thapsigargin delayed rhythms. These results suggest that the circadian clock in fibroblasts, as in SCN neurons, is regulated by membrane potential and Ca2+. Changes in intracellular Ca2+ may mediate the effects of membrane potential that we observed.
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DOI：
10.1126/science.1152506
发表时间：
2008-05-16
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
O'Neill JS;Maywood ES;Chesham JE;Takahashi JS;Hastings MH
通讯作者：
Hastings MH
影响因子：
64.5
作者：
Nitabach, MN;Blau, J;Holmes, TC
通讯作者：
Holmes, TC
影响因子：
5.3
作者：
Harrisingh, Marie C.;Wu, Ying;Nitabach, Michael N.
通讯作者：
Nitabach, Michael N.
影响因子：
--
作者：
Baxter, DF;Kirk, M;Xie, Y
通讯作者：
Xie, Y
影响因子：
64.5
作者：
Nagoshi, E;Saini, C;Schibler, U
通讯作者：
Schibler, U