My postdoctoral research focuses on investigation of genome scale transcriptional changes during axolotl limb regenration. The Mexican axolotl salamander (Ambystoma mexicanum) is unique amongst vertebrates for its remarkable regenerative capabilities. Following injury, these animals can regenerate body parts ranging from regions of the brain and heart to entire limbs and tails. While this phenomenon was first observed 250 years ago, the biological mechanism by which they accomplish such incredible feats is still relatively poorly understood. Following limb amputation, a cluster of cells known as a blastema forms at the wound site. This blastema then goes on to fully regenerate the missing limb. Previous work in our lab has identified coordinated gene expression programs that likely govern various stages of this regeneration via blastema growth. However, these experiments were performed on whole blastema samples, leaving open the critical question of just how individual cells complete such elaborately patterned tasks. Are blastemas made up of de-differentiated multipotent progenitor cells? Or, are they populated by fate-restricted migratory tissue-specific adult stem cells from stump muscle, cartilage, and blood? Is it some combination of the two? Luckily, recent advances in microfluidic technology has allowed us to address these questions. Using single cell capture platforms we have isolated and extracted total RNA of individual blastema cells from regenerating axolotl limbs. We then performed deep sequencing on these samples, yielding axolotl transcriptomes at a single cell resolution. With bioinformaticist collaborators, we are currently developing methods by which we may identify gene expression patterns that represent sub-populations of cells participating in the regeneration process over time. Ultimately, we hope that the knowledge gained through this work will someday be applied in a biomedical setting to encourage regeneration after injury or disease.
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