Germline transgenic methods for tracking cells and testing gene function during regeneration in the axolotl.
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 cell-cycle 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.
Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany ; Technische Universität Dresden, DFG Center for Regenerative Therapies, 01307 Dresden, Germany.
Shahryar
S
Khattak
Maritta
M
Schuez
Tobias
T
Richter
Dunja
D
Knapp
Saori L
SL
Haigo
Tatiana
T
Sandoval-Guzmán
Kristyna
K
Hradlikova
Annett
A
Duemmler
Ryan
R
Kerney
Elly M
EM
Tanaka
D000558:Ambystoma mexicanum; D000818:Animals; D058948:Cell Tracking / Q000379:methods*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods*; D005854:Germ Cells / Q000378:metabolism*; D018095:Germ-Line Mutation; D009416:Nerve Regeneration; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D011995:Recombination, Genetic
D019941:Cyclin-Dependent Kinase Inhibitor p16