Oliveira Catarina R, Lemaitre Regis, Murawala Prayag, Tazaki Akira, Drechsel David N, Tanaka Elly M
DFG Research Center for Regenerative Therapies, Technische Universität Dresden, Fetscherstraße 105, 01307 Dresden, Germany; Graduate Program in Areas of Basic and Applied Biology, Abel Salazar Biomedical Sciences Institute, University of Porto, 4099-003 Porto, Portugal.
Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstraße 108, 01307 Dresden, Germany.
Dev Biol. 2018 Jan 15;433(2):262-275. doi: 10.1016/j.ydbio.2017.10.008. Epub 2017 Nov 30.
Axolotls can regenerate complex structures through recruitment and remodeling of cells within mature tissues. Accessing the underlying mechanisms at a molecular resolution is crucial to understand how injury triggers regeneration and how it proceeds. However, gene transformation in adult tissues can be challenging. Here we characterize the use of pseudotyped baculovirus (BV) as an effective gene transfer method both for cells within mature limb tissue and within the blastema. These cells remain competent to participate in regeneration after transduction. We further characterize the effectiveness of BV for gene overexpression studies by overexpressing Shh in the blastema, which yields a high penetrance of classic polydactyly phenotypes. Overall, our work establishes BV as a powerful tool to access gene function in axolotl limb regeneration.
美西螈可以通过募集和重塑成熟组织中的细胞来再生复杂结构。在分子水平上了解其潜在机制对于理解损伤如何触发再生以及再生过程如何进行至关重要。然而，在成体组织中进行基因转化可能具有挑战性。在这里，我们描述了使用假型杆状病毒（BV）作为一种有效的基因转移方法，用于成熟肢体组织和芽基内的细胞。这些细胞在转导后仍有能力参与再生。我们通过在芽基中过表达Shh进一步表征了BV在基因过表达研究中的有效性，这产生了高比例的经典多指表型。总体而言，我们的工作将BV确立为研究美西螈肢体再生中基因功能的有力工具。
