Using Live-Cell Imaging and Synthetic Biology to Probe Directed Migration in Dictyostelium.

For decades, the social amoeba Dictyostelium discoideum has been an invaluable tool for dissecting the biology of eukaryotic cells. Its short growth cycle and genetic tractability make it ideal for a variety of biochemical, cell biological, and biophysical assays. Dictyostelium have been widely used as a model of eukaryotic cell motility because the signaling and mechanical networks which they use to steer and produce forward motion are highly conserved. Because these migration networks consist of hundreds of interconnected proteins, perturbing individual molecules can have subtle effects or alter cell morphology and signaling in major unpredictable ways. Therefore, to fully understand this network, we must be able to quantitatively assess the consequences of abrupt modifications. This ability will allow us better control cell migration, which is critical for development and disease, in vivo. Here, we review recent advances in imaging, synthetic biology, and computational analysis which enable researchers to tune the activity of individual molecules in single living cells and precisely measure the effects on cellular motility and signaling. We also provide practical advice and resources to assist in applying these approaches in Dictyostelium.

几十年来，社会变形虫盘基网柄菌一直是剖析真核细胞生物学的一种极有价值的工具。它较短的生长周期和易于遗传操作的特性使其非常适合各种生物化学、细胞生物学和生物物理分析。盘基网柄菌被广泛用作真核细胞运动的模型，因为它们用于引导和产生向前运动的信号传导和机械网络是高度保守的。由于这些迁移网络由数百种相互连接的蛋白质组成，干扰单个分子可能会产生微妙的影响，或者以难以预测的主要方式改变细胞形态和信号传导。因此，为了充分理解这个网络，我们必须能够定量评估突然改变的后果。这种能力将使我们能够更好地控制体内对发育和疾病至关重要的细胞迁移。在此，我们回顾了成像、合成生物学和计算分析方面的最新进展，这些进展使研究人员能够调节单个活细胞中单个分子的活性，并精确测量对细胞运动和信号传导的影响。我们还提供了实用的建议和资源，以帮助在盘基网柄菌中应用这些方法。