Department of Applied Sciences, Northumbria University, Ellison Building, City Campus, Newcastle upon Tyne, Tyne and Wear, UK.
J Mol Endocrinol. 2020 Nov;65(4):R91-R100. doi: 10.1530/JME-20-0169.
All living cells are sensors of their environment: they sense signals, hormones, cytokines, and growth factors, among others. Binding of these signals to cell surface receptors initiates the transmission of messages along intracellular signalling pathways through protein-protein interactions, enzymatic modifications and conformational changes. Typically, the activation of signalling pathways are monitored in whole populations of cells, giving population average measures, often using experimental methods that destroy and homogenise the cell population. High content imaging is an automated, high-throughput fluorescence microscopy method that enables measurements of signal transduction pathways to be taken from live cells. It can be used to measure signalling dynamics, how the abundance of particular proteins of interest change over time, or to record how particular proteins move and change their localisation in response to a signal from their environment. Using this, and other single cell methods, it is becoming increasingly clear that cells are in fact very variable in their response to a given stimulus and in the quantities of cellular components they express, even in clonal (isogenic) cell lines. This review will discuss how high content imaging has contributed to our growing understanding of cellular heterogeneity. It will discuss how data generated has been combined with information theoretic approaches to quantify the amount of information transferred through noisy signalling pathways. Lastly, the relevance of heterogeneity to our understanding and treatment of disease will be considered, highlighting the importance of single cell measurements.
它们可以感知信号、激素、细胞因子和生长因子等。这些信号与细胞表面受体结合，通过蛋白质-蛋白质相互作用、酶修饰和构象变化，启动细胞内信号通路的信息传递。通常，信号通路的激活是在细胞群体中监测的，给出群体平均测量值，通常使用破坏和均质化细胞群体的实验方法。高内涵成像（High content imaging）是一种自动化、高通量荧光显微镜方法，可从活细胞中测量信号转导通路。它可用于测量信号转导动力学，特定感兴趣的蛋白质的丰度随时间如何变化，或记录特定蛋白质如何响应来自环境的信号而移动并改变其定位。使用这种方法和其他单细胞方法，越来越明显的是，即使在克隆（同基因）细胞系中，细胞对给定刺激的反应以及它们表达的细胞成分的数量实际上是非常多样化的。本文将讨论高内涵成像如何帮助我们加深对细胞异质性的理解。它将讨论如何将生成的数据与信息论方法相结合，以量化通过噪声信号通路传递的信息量。最后，将考虑异质性对我们理解和治疗疾病的相关性，突出单细胞测量的重要性。
