Abstract：
In avascular wound repair, calcium signaling events are the predominant mechanism cells use to transduce information about stressors in the environment into an effective and coordinated migratory response. Live cell imaging and computational analysis of corneal epithelial wound healing revealed that signal initiation and propagation at the wound edge are highly ordered, with groups of cells engaging in cyclical patterns of initiation and propagation. The cells in these groups exhibit a diverse range of signaling behavior and dominant \"conductor cells\" drive activity in groups of lower-signaling neighbors. Ex vivo model systems reveal that conductor cells are present in wing cell layers of the corneal epithelium, and that signaling propagates both within and between wing and basal layers. There are significant aberrations in conductor phenotype and inter-layer propagation in Type II diabetic murine models, indicating that signal hierarchy breakdown is an early indicator of disease. In vitro models reveal that signaling profile diversity and conductor cell phenotype is eliminated with P2X7 inhibition and is altered in Pannexin-1 or P2Y2 but not Connexin-43 inhibition. Conductor cells express significantly less P2X7 than their lower-signaling neighbors and exhibit significantly less migratory behavior after injury. Together, our results show that the post-injury calcium signaling cascade exhibits significantly more ordered and hierarchical behavior than previously thought, that proteins previously shown to be essential for regulating motility are also essential for determining signaling phenotype, and that loss of signal hierarchy integrity is an early indicator of disease state.
摘要：
在无血管伤口修复中，钙信号事件是细胞用来将环境中应激源的信息转化为有效和协调的迁移反应的主要机制。角膜上皮伤口愈合的活细胞成像和计算分析表明，伤口边缘的信号起始和传播是高度有序的，细胞群参与周期性的起始和繁殖模式。这些组中的细胞表现出不同范围的信号传导行为，并且在低信号邻居组中具有优势的“导体细胞”驱动活动。离体模型系统显示，导体细胞存在于角膜上皮的翼细胞层中，信号在机翼和基底层内部和之间传播。在II型糖尿病小鼠模型中，导体表型和层间传播存在显着异常，表明信号等级分解是疾病的早期指标。体外模型显示，P2X7抑制消除了信号传导谱多样性和导体细胞表型，并在Pannexin-1或P2Y2中改变，而在连接蛋白-43抑制中没有改变。导体细胞表达的P2X7明显少于其信号传导较低的邻居，并且在损伤后表现出明显较少的迁移行为。一起,我们的结果表明，损伤后钙信号级联表现出明显更有序和分层的行为比以前认为，以前被证明是调节运动性所必需的蛋白质也是确定信号表型所必需的，信号层次完整性的丧失是疾病状态的早期指标。
