Xu Suhong, Chisholm Andrew D
Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, San Diego.
Division of Biological Sciences, Section of Cell and Developmental Biology, University of California, San Diego;
J Vis Exp. 2014 Dec 3(94):51959. doi: 10.3791/51959.
The C. elegans epidermis and cuticle form a simple yet sophisticated skin layer that can repair localized damage resulting from wounding. Studies of wound responses and repair in this model have illuminated our understanding of the cytoskeletal and genomic responses to tissue damage. The two most commonly used methods to wound the C. elegans adult skin are pricks with microinjection needles, and local laser irradiation. Needle wounding locally disrupts the cuticle, epidermis, and associated extracellular matrix, and may also damage internal tissues. Laser irradiation results in more localized damage. Wounding triggers a succession of readily assayed responses including elevated epidermal Ca(2+) (seconds-minutes), formation and closure of an actin-containing ring at the wound site (1-2 hr), elevated transcription of antimicrobial peptide genes (2-24 hr), and scar formation. Essentially all wild type adult animals survive wounding, whereas mutants defective in wound repair or other responses show decreased survival. Detailed protocols for needle and laser wounding, and assays for quantitation and visualization of wound responses and repair processes (Ca dynamics, actin dynamics, antimicrobial peptide induction, and survival) are presented.
秀丽隐杆线虫的表皮和角质层形成了一个简单却复杂的皮肤层，能够修复因创伤导致的局部损伤。对该模型中伤口反应和修复的研究，加深了我们对细胞骨架和基因组对组织损伤反应的理解。两种最常用于损伤秀丽隐杆线虫成年皮肤的方法是用显微注射针穿刺和局部激光照射。针刺会局部破坏角质层、表皮及相关的细胞外基质，还可能损伤内部组织。激光照射造成的损伤更具局部性。创伤会引发一系列易于检测的反应，包括表皮钙离子浓度升高（数秒至数分钟）、伤口部位含肌动蛋白环的形成与闭合（1至2小时）、抗菌肽基因转录升高（2至24小时）以及疤痕形成。基本上所有野生型成年动物在受伤后都能存活，而在伤口修复或其他反应方面存在缺陷的突变体存活率则会降低。本文介绍了针刺和激光损伤的详细方案，以及用于定量和可视化伤口反应及修复过程（钙离子动态变化、肌动蛋白动态变化、抗菌肽诱导和存活率）的检测方法。
