Analysis of localized cAMP perturbations within a tissue reveal the effects of a local, dynamic gap junction state on ERK signaling

Beyond natural stimuli such as growth factors and stresses, the ability to experimentally modulate at will the levels or activity of specific intracellular signaling molecule(s) in specified cells within a tissue can be a powerful tool for uncovering new regulation and tissue behaviors. Here we perturb the levels of cAMP within specific cells of an epithelial monolayer to probe the time-dynamic behavior of cell-cell communication protocols implemented by the cAMP/PKA pathway and its coupling to the ERK pathway. The time-dependent ERK responses we observe in the perturbed cells for spatially uniform cAMP perturbations (all cells) can be very different from those due to spatially localized perturbations (a few cells). Through a combination of pharmacological and genetic perturbations, signal analysis, and computational modeling, we infer how intracellular regulation and regulated cell-cell coupling each impact the intracellular ERK response in single cells. Our approach reveals how a dyna...

除生长因子、细胞应激等天然刺激因素外，能够在组织内的特定细胞中自由实验性调控特定细胞内信号分子（intracellular signaling molecule）的水平或活性，可成为揭示全新调控机制与组织行为的有力工具。本研究通过调控上皮单层（epithelial monolayer）中特定细胞内的环腺苷酸（cyclic adenosine monophosphate, cAMP）水平，旨在探究由cAMP/蛋白激酶A（protein kinase A, PKA）通路介导的细胞间通讯的时间动态特征，以及该通路与细胞外调节蛋白激酶（extracellular regulated protein kinases, ERK）通路的耦合机制。我们观察到，当对所有细胞施加空间均匀的cAMP扰动时，受扰细胞内的ERK响应随时间的变化特征，与仅对少量细胞施加空间局部扰动时的响应存在显著差异。本研究结合药理学扰动、遗传扰动、信号分析与计算建模手段，解析了细胞内调控机制与受调控的细胞间耦合分别如何影响单个细胞内的ERK响应。本研究方法揭示了动态（dynamic）[原文未完结]