Featherstone Karen, Hey Kirsty, Momiji Hiroshi, McNamara Anne V, Patist Amanda L, Woodburn Joanna, Spiller David G, Christian Helen C, McNeilly Alan S, Mullins John J, Finkenstädt Bärbel F, Rand David A, White Michael R H, Davis Julian R E
Centre for Endocrinology and Diabetes, University of Manchester, Manchester, United Kingdom.
Department of Statistics, University of Warwick, Coventry, United Kingdom.
Elife. 2016 Feb 1;5:e08494. doi: 10.7554/eLife.08494.
Transcription at individual genes in single cells is often pulsatile and stochastic. A key question emerges regarding how this behaviour contributes to tissue phenotype, but it has been a challenge to quantitatively analyse this in living cells over time, as opposed to studying snap-shots of gene expression state. We have used imaging of reporter gene expression to track transcription in living pituitary tissue. We integrated live-cell imaging data with statistical modelling for quantitative real-time estimation of the timing of switching between transcriptional states across a whole tissue. Multiple levels of transcription rate were identified, indicating that gene expression is not a simple binary 'on-off' process. Immature tissue displayed shorter durations of high-expressing states than the adult. In adult pituitary tissue, direct cell contacts involving gap junctions allowed local spatial coordination of prolactin gene expression. Our findings identify how heterogeneous transcriptional dynamics of single cells may contribute to overall tissue behaviour.
单细胞中单个基因的转录通常是脉动性和随机性的。一个关键问题出现了,即这种行为如何影响组织表型,但与研究基因表达状态的快照不同,随着时间的推移对活细胞进行定量分析一直是一项挑战。我们利用报告基因表达成像来追踪活垂体组织中的转录过程。我们将活细胞成像数据与统计模型相结合,以实时定量估计整个组织中转录状态之间切换的时间。我们确定了多个转录速率水平,这表明基因表达不是一个简单的二元“开-关”过程。未成熟组织中高表达状态的持续时间比成体组织短。在成体垂体组织中,涉及缝隙连接的直接细胞接触允许催乳素基因表达进行局部空间协调。我们的研究结果揭示了单细胞的异质转录动态如何可能影响整体组织行为。