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- W3188656584 abstract "Ambystoma mexicanum, or the Mexican axolotl is one of the few tetrapod vertebrates which exhibits the ability to perform complex tissue regeneration throughout its life. While humans and other mammals do not share this amazing regenerative capacity, it is the hope that through studying this process in regeneratively competent vertebrates that we can inform translational studies and future therapies. Our work details the implementation and optimization of new methodologies for the investigation of gene expression during the regenerative process of the axolotl. We have detailed the use of a new in-situ staining methodology, HCR FISH, for visualizing multiple RNA species simultaneously in both spinal cord and limb tissue. This method has shown promise in not only detailing the expression patterns within tissue, but also the characterization of cell populations based on similarities in gene expression through relative quantification of observed expression. Additionally, we describe the role of Wnt signaling, a key developmental pathway, in the regeneration on the axolotl limb. We observed Wnt5a, 5b, and 3a expression within the blastema and AEC of the regenerating limb. To functionally test the role of Wnt we pharmacologically inhibited Wnt ligand secretion, and in response saw a decrease in regenerative ability. We also observed a decrease in expression of fgf8 and shh, 2 genes integral for promoting cell proliferation and patterning, in Wnt inhibited limbs when compared to untreated. We attempted to rescue the inhibition of Wnt secretion through activation of b-catenin signaling and observed that this rescue was no different than the limbs treated with only the inhibiter. These findings suggest that Wnt expression during regeneration play key role in promoting cell proliferation and outgrowth, and hints at the functionality of Wnt signaling in regeneration is not limited to a single Wnt pathway. --Author's abstract" @default.
- W3188656584 created "2021-08-16" @default.
- W3188656584 creator A5026676778 @default.
- W3188656584 date "2021-05-10" @default.
- W3188656584 modified "2023-09-25" @default.
- W3188656584 title "Implementation of multiplexed fluorescence in situ hybridization for studying regeneration in the axolotl salamander" @default.
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