MOLECULAR GENETIC ANALYSIS OF WOUND HEALING--DROSOPHILA

导致组织或组织完整性丧失的创伤构成伤口，并启动伤口愈合或修复过程。该过程的主要特征包括实现组织的即时稳态，排斥微生物，并最终恢复正常的组织生理机能。人类的创伤和伤口愈合具有重要的生物学和社会学意义。伤口是意外造成的，可能是在愤怒中造成的，也可能是在手术中由于有益的医学原因造成的。我们提出的工作的最终目标是了解上皮细胞对伤口反应的基本生物学，并为设计旨在加速愈合或防止过度瘢痕形成的干预策略做出贡献。许多脊椎动物，特别是哺乳动物，动物模型系统已经发展到研究伤口和伤口愈合。因此，负责适当伤口愈合的分子角色已经开始被确定。近年来，基因敲除和转基因小鼠技术已被应用于伤口愈合的研究。然而，哺乳动物模型系统只能在经过仔细的伦理考虑后才能用于伤害研究，这自然是耗时且非常昂贵的。此外，前向基因筛选基本上是不切实际的。在这里，我们建议追求分子/遗传策略，这是在果蝇中唯一可用的，以研究损伤和伤口愈合的生物学。最近，我们使用机械和激光方法消融果蝇胚胎的组织，并发现这种消融之后是快速和强大的愈合。此外，我们还发现，由于肌球蛋白重链结构基因的缺陷，胚胎的愈合完全受到抑制。我们建议对果蝇的损伤和伤口愈合进行表征，测试候选基因座在损伤和伤口愈合过程中的作用，并进行前向遗传筛选以恢复无法正常愈合的基因座。这些研究将直接解决上皮伤口愈合的那些方面，这些方面保存了5到10亿年的进化，将昆虫与哺乳动物区分开来。他们承诺将利用果蝇遗传学和分子遗传学的强大工具来解决这一重要的生物学问题。

Trauma that results in loss of tissue or tissue integrity constitutes a wound, and initiates a wound healing or repair process. Key features of that process include achieving immediate tissue homeostasis, repelling microorganisms and ultimately, the restoration of normal tissue physiology. Wounding and wound healing in humans is of considerable biological and social significance. Wounds occur by accident, can be inflicted in anger, or occur during surgery, for beneficial medical reasons. The ultimate goal of the work we propose is to understand the basic biology of epithelial response to wounds with the long range goal of contributing to the design of intervention strategies designed to speed healing or prevent excessive scarring. A number of vertebrate, and especially mammalian, animal model systems have been developed to study wounding and wound healing. As a consequence, the molecular players responsible for proper wound healing have begun to be identified. Recently, knockout and transgenic mouse technologies have been applied to the study of wound healing. Nevertheless, mammalian model systems can be used in wounding studies only after careful ethical consideration, are naturally time consuming and are very expensive. Moreover, forward genetic screens are essentially impractical. Here, we propose to pursue molecular/genetic strategies that are uniquely available in Drosophila to investigate the biology of wounding and wound healing. Recently we used mechanical and laser methods to ablate tissue in Drosophila embryos and found that such ablations are followed by rapid and robust healing. Moreover, we found that healing was completely inhibited in embryos homozygous for defects in the structural gene for myosin heavy chain. We propose to characterize wounding and wound healing in Drosophila, test the role of candidate loci in the process of wounding and wound healing, and perform forward genetic screens to recover loci that fail to heal properly. These studies will address directly those aspects of epithelial wound healing that are conserved over the 0.5 to 1 billion years of evolution that separate insects from mammals. Together, they promise to bring the powerful tools of Drosophila genetics and molecular genetics to bear on this important biological problem.