Author ORCID Identifier

https://orcid.org/0000-0003-1496-2561

Year of Publication

2019

Degree Name

Doctor of Philosophy (PhD)

Document Type

Doctoral Dissertation

College

Arts and Sciences

Department

Biology

First Advisor

Dr. S. Randal Voss

Abstract

Salamanders have an extraordinary ability to regenerate appendages after loss or amputation, irrespective of age. My dissertation research explored the possibility that regenerative ability is associated with the evolution of novel, salamander-specific genes. I utilized transcriptional and genomic databases for the axolotl to discover previously unidentified genes, to the exclusion of other vertebrate taxa. Among the genes identified were multiple mmps (Matrix metalloproteases) and a jnk1/mapk8 (c-jun-N-terminal kinase) paralog. MMPs function in extracellular matrix remodeling (ECM) and tissue histolysis, processes that are essential for successful regeneration. Jjnk1/mapk8 plays a pivotal role in regulating transcription in response to cellular stress stimuli, including ROS (reactive oxygen species). Discovery of these novel genes motivated further bioinformatic studies of mmps and wet-lab experiments to characterize JNK and ROS signaling. The paralogy of the newly discovered mmps and orthology of 15 additional mmps was established by analyses of predicted, protein secondary structures and gene phylogeny. A microarray-analysis identified target genes downstream of JNK signaling that are predicted to function in cell proliferation, cellular stress response, and ROS production. These inferences were validated by additional experiments that showed a requirement for NOX (NADPH oxidase) activity, and thus presumably ROS production for successful tail regeneration. In summary, my dissertation identified novel, salamander-specific genes. The functions of these genes suggest that regenerative ability is associated with a diverse extracellular matrix remodeling and/or tissue histolysis response, and also stress-associated signaling pathways. The bioinformatic findings and functional assays that were developed to quantify ROS, cell proliferation, and mitosis will greatly empower the axolotl embryo model for tail regeneration research.

Digital Object Identifier (DOI)

https://doi.org/10.13023/etd.2019.470

Funding Information

This research was supported by National Institutes of Health (NIH): R24OD021479 and P40OD01979. Through 2017-2019.

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