The Role of Sustained Calcium Oscillations in the Wound Healing Response

项目摘要 角膜是眼睛最前面的结构，这使得它很容易被抓伤， 感染和营养不良。事实上，由于受伤和疾病导致的角膜失明是导致 全球范围内可预防的失明。这种情况的唯一根治方法是角膜移植或 移植，但可存活的眼睛组织并不是在全球范围内就能买到。因此，研究黄曲霉毒素的作用机制。 角膜伤口的正常愈合可以为我们提供潜在的新治疗方法的发展信息 目标。损伤后，角膜上皮细胞释放核苷酸(如三磷酸腺苷)激活嘌呤能受体 P2Y2是G蛋白偶联受体，而P2X7是配体门控离子通道。两个都是 这些受体诱导一过性钙波，这已被证实是伤口中的关键步骤。 治愈的过程。然而，人们对钙信号在伤口修复中的长期作用知之甚少。vt.给出 我们推测，在最初的钙波之后，P2X7和P2Y2的表达和定位发生了变化 这些受体也可能参与伤口愈合过程的后期。我们现在有了 确定了钙传播的长期作用，当被抑制时，会减弱愈合过程。这 次要反应发生在原发钙波诱导后10-30分钟，包括 在多个细胞簇内传播的一系列钙振荡，并被认为持续了几个月 几个小时。用胞外核糖核酸酶使胞外核苷酸失活会抑制二次反应， 这表明在这个过程中需要P2Y2和P2X7。与主要响应不同，次要响应 钙反应依赖于细胞间的接触，因此受亚汇流条件和缝隙的抑制。 连接抑制药。这些结果表明，创伤后的钙反应比创伤后的钙反应复杂。 两个受体中的任何一个单独。我这个项目的目标是研究嘌呤能受体P2X7如何 和P2Y2协调和产生次级钙反应以及这种反应是如何起作用的 对伤口愈合过程的影响。因此，我假设嘌呤能受体P2X7和P2Y2 以不同的方式调节二次钙反应，参与伤口修复的多个过程。

PROJECT ABSTRACT The cornea is the most anterior structure of the eye, which makes it susceptible to injury from scratching, infections, and dystrophy. In fact, corneal blindness from injury and disease is the fourth highest cause of preventable blindness globally. The only curative treatment for this condition is corneal transplantation or grafting, but viable eye tissues are not readily available worldwide. Therefore, studying the mechanism of normal wound healing in the cornea could give us information on development of potential novel therapeutic targets. Upon injury, corneal epithelial cells release nucleotides (e.g. ATP) that activate purinergic receptors P2Y2 and P2X7; P2Y2 is a G-protein coupled receptor, while P2X7 is a ligand-gated ion channel. Both of these receptors induce a transient calcium wave that has been established to be a critical step in the wound healing process. However, little is known about the long-term role of calcium signaling in wound repair. Given that the expression and localization of P2X7 and P2Y2 change after the initial calcium wave, we speculated that these receptors may also be involved in the later stages of the wound healing process. We have now identified a long-term role of calcium propagation that when inhibited attenuates the healing process. This secondary response, which occurs 10-30 minutes after the primary calcium wave is induced, consists of a series of calcium oscillations that propagate within multiple clusters of cells, and it was seen to last for several hours. Deactivating extracellular nucleotides with ectonucleotidases inhibited the secondary response, indicating that P2Y2 and P2X7 are required in this process. Unlike the primary response, the secondary calcium response depended on cell-cell contact and thus was inhibited by sub-confluent conditions and gap junction inhibitors. These results demonstrate that the calcium response after wounding is more complex than that of either receptor alone. The goal of my project is to investigate how the purinergic receptors P2X7 and P2Y2 coordinate and produce the secondary calcium response and how that response contributes to the wound healing process. Therefore, I hypothesize that purinergic receptors P2X7 and P2Y2 differentially mediate the secondary calcium response and contribute to multiple processes in wound repair.