Germline Transgenic Methods for Tracking Cells and Testing Gene Function during Regeneration in the Axolotl
Germline Transgenic Methods for Tracking Cells and Testing Gene Function during Regeneration in the Axolotl
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DOI：
10.1016/j.stemcr.2013.03.002
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发表时间：
2013-06-04
影响因子：
5.9
通讯作者：
Tanaka, Elly M.
中科院分区：
文献类型：
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作者：
Khattak, Shahryar;Schuez, Maritta;Tanaka, Elly M.
蝾螈是唯一一种可以终生再生复杂身体结构的四足动物。破解再生的潜在分子过程是再生医学和发育生物学的基础，但模式生物的分子分析工具有限。我们描述了实验室培育的蝾螈Ambystoma mexicanum （axolotl）的一套全面的种系转基因菌株，开辟了再生的细胞和分子遗传学解剖。我们证明了神经、雪旺细胞、少突胶质细胞、肌肉、表皮和软骨中基因表达的组织依赖性控制。此外，我们证明了使用他莫昔芬诱导的Cre/ loxp介导的重组来不可磨灭地标记不同的细胞类型。最后，我们诱导过表达细胞周期抑制剂p16(INK4a)，它负向调节脊髓再生。这些组织特异性的种系蝾螈系和紧密诱导的Cre驱动和LoxP报告系使这种经典的再生模型在分子上可实现。
The salamander is the only tetrapod that regenerates complex body structures throughout life. Deciphering the underlying molecular processes of regeneration is fundamental for regenerative medicine and developmental biology, but the model organism had limited tools for molecular analysis. We describe a comprehensive set of germline transgenic strains in the laboratory-bred salamander Ambystoma mexicanum (axolotl) that open up the cellular and molecular genetic dissection of regeneration. We demonstrate tissue-dependent control of gene expression in nerve, Schwann cells, oligodendrocytes, muscle, epidermis, and cartilage. Furthermore, we demonstrate the use of tamoxifen-induced Cre/loxP-mediated recombination to indelibly mark different cell types. Finally, we inducibly overexpress the cellcycle inhibitor p16(INK4a), which negatively regulates spinal cord regeneration. These tissue-specific germline axolotl lines and tightly inducible Cre drivers and LoxP reporter lines render this classical regeneration model molecularly accessible.