Molecular generators at corneal wounds produce and regulate the wound electrical signals

描述（由申请人提供）：我们的目标是电愈合伤口。我们和其他实验室的实验表明，电场在角膜伤口处自然产生，并提供强大的信号来引导角膜上皮细胞的迁移。我们证明了电信号可以超越其他方向线索，如损伤刺激、自由边缘和机械力，来指导角膜上皮片和大群细胞的迁移。然而，目前尚不清楚角膜伤口是如何产生电信号的。我们建议在角膜伤口处发现分子“发生器”。发现这些发生器分子将给我们提供分子目标，我们可以刺激它们产生更强的电信号，从而促进愈合。在这项提议中，我们将1)利用电生理学和遗传模型表征角膜上皮细胞中的钙活化氯离子通道（CaCCs）；2)确定CaCCs对损伤的反应及其在产生伤口电流中的作用；3)纠正糖尿病大鼠和ANO1基因敲除小鼠有缺陷的创面电信号（和创面愈合）。我们期望发现的新的机制将为慢性角膜创伤的愈合提供基本的见解和实用的策略。

DESCRIPTION (provided by applicant): Our goal is to electrically heal wounds. Experiments from our own lab and others have demonstrated that electric fields occur naturally at corneal wounds and provide a powerful signal to guide migration of corneal epithelial cells. We demonstrated that the electrical signals could override other directional cues, such as injury stimulation, free edge and mechanical forces, in guiding migration of corneal epithelial sheets and large groups of cells. It is however not known how cornea wounds generate the electric "signal". We propose to discover molecular "generators" at corneal wounds. Uncovering these generator molecules will give us molecular targets which we can stimulate to produce stronger electrical signals, and thus facilitate healing. In this proposal, we will 1) characterize calcium-activated chloride channels (CaCCs) in corneal epithelial cells using electrophysiology and genetic models; 2) determine responses of CaCCs to injury and their roles in generating wound electric currents; and, 3) correct the defective wound electric signals (and wound healing) in diabetic rats and ANO1 knockout mice. We expect that the novel mechanisms discovered will provide fundamental insights as well as practical strategies for healing of chronic corneal wounds.