Abstract
Regeneration of fish fins and urodele limbs occurs via formation of the blastema, which is a mass of mesenchymal cells formed at the amputated site and is essential for regeneration. The blastema transplantation, a novel technique developed in our previous studies (Shibata et al., 2016; Yoshinari et al., 2012) is a useful approach for tracking and manipulating the blastema cells during fish fin regeneration.
Keywords: Zebrafish, Fin regeneration, Blastema, Transplantation, Cell lineage
Background
Cell transplantation studies are routinely performed during the early embryonic stage in animal models such as fish, amphibians and mammals, but targeting the transplanted cells to specific tissues has been difficult. Blastema transplantation developed in our studies is an efficient method for introducing mesenchymal donor cells into host fin ray. It enables us to track cell fate and/or manipulate cell signaling such as fibroblast growth factor (Fgf) during fish fin regeneration. Actually, in our recently published work, we transplanted blastema cells, which carried the hsp70l:dominant-negative fgf receptor and the β-actin:dsRed2 transgenes, into a wild-type blastema region and mosaically inhibited Fgf signaling in a subset of fin ray mesenchymal cells (Shibata et al., 2016). This method is applicable for analyzing other cell signals and for tracking cell fate by live cell imaging.
Materials and Reagents
Equipment
Procedure
Recipes
Acknowledgments
This protocol was adapted from our previous works (Yoshinari et al., 2012; Shibata et al., 2016). The works were supported by the Grant-in-Aid for Scientific Research from Japan Society for the Promotion of Science (JSPS).
References
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