Katsuyama Tomonori, Paro Renato
Department of Genetics, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo, 113-0033, Japan.
Department of Biosystems Science and Engineering, Eidgenössische Technische Hochschule (ETH) Zürich, Mattenstrasse 26, 4058, Basel, Switzerland.
Methods Mol Biol. 2016;1480:301-10. doi: 10.1007/978-1-4939-6380-5_26.
Since Ephrussi and Beadle introduced imaginal disc transplantation to Drosophila research in 1936, the method played an important part towards a better understanding of disc patterning, tissue regeneration, and reprogramming phenomena like transdetermination. Despite increasing usage of high-throughput approaches towards solving biological problems this classical manual method is still in use for studying disc development in a semi-physiological context. Here we describe in detail a protocol and provide recommendations on the procedure in particular for analyzing the regenerative potential of imaginal disks. The steps consist of disc dissection and fragmentation, transplantation into the larval or adult abdomen, and the recovery of implants from the host abdomen. Additionally, we also describe how to make the special transplantation needle from a glass capillary.
自1936年埃弗鲁西和比德尔将成虫盘移植引入果蝇研究以来,该方法在更好地理解盘模式形成、组织再生以及诸如转决定等重编程现象方面发挥了重要作用。尽管解决生物学问题的高通量方法的使用日益增加,但这种经典的手工方法仍在用于在半生理背景下研究盘发育。在这里,我们详细描述了一种方案,并针对该程序提供了建议,特别是用于分析成虫盘的再生潜力。步骤包括盘解剖和破碎、移植到幼虫或成虫腹部以及从宿主腹部回收植入物。此外,我们还描述了如何用玻璃毛细管制作特殊的移植针。