Monitoring the regulation of gene expression in a growing organ using a fluid mechanics formalism.
Monitoring the regulation of gene expression in a growing organ using a fluid mechanics formalism.
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DOI：
10.1186/1741-7007-8-18
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发表时间：
2010-03-04
期刊：
影响因子：
5.4
通讯作者：
Bogeat-Triboulot MB
中科院分区：
文献类型：
--
作者：
Merret R;Moulia B;Hummel I;Cohen D;Dreyer E;Bogeat-Triboulot MB
技术进步使得基因表达的准确定量成为可能，即使是在单细胞类型中。虽然常规进行转录组分析，但大多数实验设计仅提供基因表达的快照。通过这些不连续的数据集，揭示了细胞命运或位置信号的分子机制。然而，在发展多细胞结构时，已知直接影响转录网络的时间和空间线索会随着细胞的移位和扩张而纠缠在一起。获得一个公正的观点的时空调控发生在发展过程中的基因表达需要一个特定的框架，适当量化的变化率的属性在一个移动和扩展的元素，如细胞或器官节段。我们展示了如何通过结合运动学和实时聚合酶链反应数据在一个机械模型，认为任何器官作为一个连续体的基因表达的变化率可以量化。应用该框架研究了Actin 11和ElongationFactor 1-β两个参比基因对白杨根尖发育的调控作用。通过延时摄影确定生长场，并以高空间分辨率获得转录物密度。这两个基因的转录本的净积累率被发现显示出高度对比的发展概况。Actin 11在生长带的加速区和减速区表现出脉冲式的上下调节，而EF 1 β的动态变化则慢得多。该框架提供了关于发育器官中基因调控的关键信息，例如基因诱导/抑制的位置、持续时间和强度。我们证明，基因表达模式可以监测使用的连续性方程，而不使用突变体或报告结构。鉴于成像技术的兴起，我们认为这一框架为剖析生长调控的分子基础开辟了一条新途径，即使是在非模型物种或复杂结构中。
Technological advances have enabled the accurate quantification of gene expression, even within single cell types. While transcriptome analyses are routinely performed, most experimental designs only provide snapshots of gene expression. Molecular mechanisms underlying cell fate or positional signalling have been revealed through these discontinuous datasets. However, in developing multicellular structures, temporal and spatial cues, known to directly influence transcriptional networks, get entangled as the cells are displaced and expand. Access to an unbiased view of the spatiotemporal regulation of gene expression occurring during development requires a specific framework that properly quantifies the rate of change of a property in a moving and expanding element, such as a cell or an organ segment. We show how the rate of change in gene expression can be quantified by combining kinematics and real-time polymerase chain reaction data in a mechanistic model which considers any organ as a continuum. This framework was applied in order to assess the developmental regulation of the two reference genes Actin11 and Elongation Factor 1-β in the apex of poplar root. The growth field was determined by time-lapse photography and transcript density was obtained at high spatial resolution. The net accumulation rates of the transcripts of the two genes were found to display highly contrasted developmental profiles. Actin11 showed pulses of up and down regulation in the accelerating and decelerating parts of the growth zone while the dynamic of EF1β were much slower. This framework provides key information about gene regulation in a developing organ, such as the location, the duration and the intensity of gene induction/repression. We demonstrated that gene expression patterns can be monitored using the continuity equation without using mutants or reporter constructions. Given the rise of imaging technologies, this framework in our view opens a new way to dissect the molecular basis of growth regulation, even in non-model species or complex structures.
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影响因子：
5.1
作者：
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