Phase-locked signals elucidate circuit architecture of an oscillatory pathway.
Phase-locked signals elucidate circuit architecture of an oscillatory pathway.
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DOI：
10.1371/journal.pcbi.1001040
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发表时间：
2010-12-23
影响因子：
4.3
通讯作者：
Takayama S
中科院分区：
文献类型：
--
作者：
Jovic A;Howell B;Cote M;Wade SM;Mehta K;Miyawaki A;Neubig RR;Linderman JJ;Takayama S
本文介绍了振荡细胞信号系统的锁相分析的概念，阐明生化电路结构。锁相是一种物理现象，指的是系统输出与周期性刺激同步的响应模式;在某些情况下，响应的数量可能少于输入的数量，指示跳过的节拍。虽然单独观察锁相在很大程度上是独立的详细机制，我们发现，锁相的属性是有用的区分电路架构，因为它们不仅反映了激活，但也生化电路的恢复特性。在这里，这一原则被证明为分析的G蛋白偶联受体系统，M3毒蕈碱受体-钙信号通路，使用微流体介导的周期性化学刺激的M3受体与卡巴胆碱和实时成像所产生的钙瞬变。使用这种方法，我们发现了基础IP 3生产的潜在重要性，这一发现对钙响应保真度周期性刺激具有重要意义。基于我们的分析，我们还否定了Gq-PLC相互作用是开关样的概念，这对细胞外信号如何被过滤和下游解释有很大影响。锁相分析是一种新的和有用的工具，模型的修订和机制阐明，该方法补充了传统的遗传和化学工具的细胞信号通路的分析，并应广泛适用于其他振荡途径。对单元电路架构进行鲁棒识别的关键是具有尽可能多的不同响应特征以用于比较和评估。振荡电路的一个未被充分认识的特性是，在周期性刺激下，这些系统将表现出与该刺激输入同步的响应，这种现象称为锁相。我们表明，锁相响应特性的变化显着取决于电路的激活和恢复性能，这些响应特性，从而提供了一套独特的标准振荡电路架构分析。这一概念通过哺乳动物细胞中振荡钙通道的实验得到验证;实验装置使我们能够首次探索细胞内信号化学诱导锁相的特性。然后，对这种现象的观察被用来测试几个现有的钙信号数学模型的预测。我们评估的大多数模型都无法匹配我们所有的实验观察结果，这表明目前的模型在测试的细胞类型和受体/兴奋剂的钙信号传导背景下缺少机械元素。锁相的观察进一步引导我们确定了一个简单的机制修改，可以解释所有的实验观察。所提出的技术和方法应广泛适用于各种生物振荡器。
This paper introduces the concept of phase-locking analysis of oscillatory cellular signaling systems to elucidate biochemical circuit architecture. Phase-locking is a physical phenomenon that refers to a response mode in which system output is synchronized to a periodic stimulus; in some instances, the number of responses can be fewer than the number of inputs, indicative of skipped beats. While the observation of phase-locking alone is largely independent of detailed mechanism, we find that the properties of phase-locking are useful for discriminating circuit architectures because they reflect not only the activation but also the recovery characteristics of biochemical circuits. Here, this principle is demonstrated for analysis of a G-protein coupled receptor system, the M3 muscarinic receptor-calcium signaling pathway, using microfluidic-mediated periodic chemical stimulation of the M3 receptor with carbachol and real-time imaging of resulting calcium transients. Using this approach we uncovered the potential importance of basal IP3 production, a finding that has important implications on calcium response fidelity to periodic stimulation. Based upon our analysis, we also negated the notion that the Gq-PLC interaction is switch-like, which has a strong influence upon how extracellular signals are filtered and interpreted downstream. Phase-locking analysis is a new and useful tool for model revision and mechanism elucidation; the method complements conventional genetic and chemical tools for analysis of cellular signaling circuitry and should be broadly applicable to other oscillatory pathways. Key to robust discernment of cell circuit architecture is to have as many distinct response features as possible for comparison and evaluation. One under-appreciated characteristic of oscillatory circuits is that under periodic stimulation, these systems will exhibit responses synchronized to this stimulatory input, a phenomenon termed phase-locking. We demonstrate that phase-locked response characteristics vary noticeably depending on circuit activation and recovery properties; these response characteristics thereby provide a unique set of criteria for oscillatory circuit architecture analysis. The concept is validated through experiments on an oscillatory calcium pathway in mammalian cells; the experimental setup allowed us to explore, for the first time, the properties of chemically induced phase-locking of intracellular signals. Observations of this phenomenon were then used to test the predictions of several existing mathematical models of calcium signaling. Most of the models we evaluated were unable to match all our experimental observations, suggesting that current models are missing mechanistic elements in the context of calcium signaling for the cell type and receptor/stimulant tested. The observations of phase-locking further led us to identify one simple mechanistic modification that would account for all the experimental observations. The techniques and methodology presented should be broadly applicable to a variety of biological oscillators.
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DOI：
10.1098/rspb.1995.0153
发表时间：
1995-09-22
影响因子：
4.7
作者：
GOLDBETER, A
通讯作者：
GOLDBETER, A
影响因子：
7.4
作者：
Heo, Yun Seok;Cabrera, Lourdes M.;Takayama, Shuichi
通讯作者：
Takayama, Shuichi
影响因子：
4
作者：
CUTHBERTSON, KSR;CHAY, TR
通讯作者：
CHAY, TR
DOI：
10.1073/pnas.0404353101
发表时间：
2004-11-09
影响因子：
11.1
作者：
Gu, W;Zhu, XY;Takayama, S
通讯作者：
Takayama, S
影响因子：
4.3
作者：
Kinzer-Ursem TL;Linderman JJ
通讯作者：
Linderman JJ