Satellite cells are adult muscle stem cells that activate, proliferate, and differentiate into myofibers upon muscle damage. Satellite cells can be cultured and manipulated in vitro, and thus represent an accessible model for studying skeletal muscle biology, and a potential source of autologous stem cells for regenerative medicine. This work summarizes efforts to further understanding of satellite cell biology, using novel model organisms, bioengineering, and molecular and cellular approaches. Lizards are evolutionarily the closest vertebrates to humans that regenerate entire appendages. An analysis of lizard myoprogenitor cell transcriptome determined they were most transcriptionally similar to mammalian satellite cells. Further examination showed that among genes with the highest level of expression in lizard satellite cells were an increased number of regulators of chondrogenesis. In micromass culture, lizard satellite cells formed nodules that expressed chondrogenic regulatory genes, thus demonstrating increased musculoskeletal plasticity. However, to exploit satellite cells for therapeutics, development of an ex vivo culture is necessary. This work investigates whether substrates composed of extracellular matrix (ECM) proteins, as either coatings or hydrogels, can support expansion of this population whilst maintaining their myogenic potency. Stiffer substrates are necessary for in vitro proliferation and differentiation of satellite cells, while the ECM composition was not significantly important. Additionally, satellite cells on hydrogels entered a quiescent state that could be reversed when the cells were subsequently cultured on Matrigel. Proliferation and gene expression data further indicated that C2C12 cells are not a good proxy for satellite cells. To further understand how different signaling pathways control satellite cell behavior, an investigation of the Notch inhibitor protein Numb was carried out. Numb deficient satellite cells fail to activate, proliferate and participate in muscle repair. Examination of Numb isoform expression in satellite cells and embryonic tissues revealed that while developing limb bud, neural tube, and heart express the long and short isoforms of NUMB, satellite cells predominantly express the short isoforms. A preliminary immunoprecipitation- proteomics experiment suggested that the roles of NUMB in satellite cells are related to cell cycle modulation, cytoskeleton dynamics, and regulation of transcription factors necessary for satellite cell function.

Traumatic injury to the central nervous or musculoskeletal system in traditional amniote models, such as mouse and chicken, is permanent with long-term physiological and functional effects. However, among amniotes, the ability to regrow complex, multi-tissue structures is unique to non-avian reptiles. Structural regeneration is extensively studied in lizards, with most species able to regrow a functional tail. The lizard regenerated tail includes the spinal cord, cartilage, de novo muscle, vasculature, and skin, and unlike mammals, these tissues can be replaced in lizards as adults. These studies focus on the events that occur before and after the tail regrowth phase, identifying conserved mechanisms that enable functional tail regeneration in the green anole lizard, Anolis carolinensis. An examination of coordinated interactions between peripheral nerves, Schwann cells, and skeletal muscle reveal that reformation of the lizard neuromuscular system is dependent upon developmental programs as well as those unique to the adult during late stages of regeneration. On the other hand, transcriptomic analysis of the early injury response identified many immunoregulatory genes that may be essential for inhibiting fibrosis and initiating regenerative programs. Lastly, an anatomical and histological study of regrown alligator tails reveal that regenerative capacity varies between different reptile groups, providing comparative opportunities within amniotes and across vertebrates. In order to identify mechanisms that limit regeneration, these cross-species analyses will be critical. Taken together, these studies serve as a foundation for future experimental work that will reveal the interplay between reparative and regenerative mechanisms in adult amniotes with translational implications for medical therapies.

New genomic resources allow for the investigation of gene family diversity in genome-enabled reptiles. The Toll-like Receptor (TLR) gene family recognizes pathogen-associated molecular patterns (PAMPs) and coevolves with environmental pathogens which makes it a strong candidate for looking at the interplay between gene family diversification and host-pathogen coevolution. Using a new orthology curation pipeline and phylogenetic reconstruction, a novel gene expansion event of TLR8 was identified to be exclusive to crocodilians and chelonians with species-specific pseudogenization events. A new gene, TLR21-like, was identified as a part of the TLR11 subfamily. These findings uncover reptile-specific gene family evolution and provide indications of the role of habitat in this process.

Vitellogenin (Vg) is an ancient and highly conserved multifunctional protein. It is primarily known for its role in egg-yolk formation but also serves functions pertaining to immunity, longevity, nutrient storage, and oxidative stress relief. In the honey bee (Apis mellifera), Vg has evolved still further to include important social functions that are critical to the maintenance and proliferation of colonies. Here, Vg is used to synthesize royal jelly, a glandular secretion produced by a subset of the worker caste that is fed to the queen and young larvae and which is essential for caste development and social immunity. Moreover, Vg in the worker caste sets the pace of their behavioral development as they transition between different tasks throughout their life. In this dissertation, I make several new discoveries about Vg functionality. First, I uncover a colony-level immune pathway in bees that uses royal jelly as a vehicle to transfer pathogen fragments between nestmates. Second, I show that Vg is localized and expressed in the honey bee digestive tract and suggest possible immunological functions it may be performing there. Finally, I show that Vg enters to nucleus and binds to deoxyribonucleic acid (DNA), acting as a potential transcription factor to regulate expression of many genes pertaining to behavior, metabolism, and signal transduction pathways. These findings represent a significant advance in the understanding of Vg functionality and honey bee biology, and set the stage for many future avenues of research.

We propose the Bio-HCI framework, that focuses on three major components: biological materials, intermediate platforms, and interaction with the user. In this context, "biological materials" is meant to broadly cover biological matter (DNA, RNA, enzyme), biological information (gene, epigenetic), biological process (mutation, reproduction, self assembling), and biological form. These biological materials serve as the design elements for designers to use in the same way as digital materials. Intermediate Platform focuses on methods of connecting biological materials to a user, or a digital platform that connect to users. In most current use-cases, biological materials need an intermediate platform to transfer the information to the user and transfer the user's response back to biological materials. Examples include a DNA sequencer, microscope, or petri dish. User interaction emphasizes the interactivity between a user and the biological machine (biological materials + intermediate platform). The interaction ranges from a basic human-computer interaction such as using a biological machine as a file storage to a unique interaction such as having a biological machine that evolves to solve user's task. To examine this framework further, we present four experiments which focus on the different aspect of the Bio-HCI framework.

The modern tetraploid species Gossypium barbadense L. (AD2) traces its origins to an allopolyploidy event between diploid progenitors G. raimondii (DT Genome, Americas) and G. herbaceum (AT Genome, Asia/Africa). In this study, nine fiber-related genes consisting of seven MYB transcription factors, a cellulose synthase homolog, and a tubulin homolog were resequenced across 54 G. barbadense lines spanning the wild-to-domesticated spectrum. Tests for nucleotide diversity (π), linkage disequilibrium (LD), and Tajima’s D were performed to examine the extent to which evolutionary forces have acted on these nine loci in G. barbadense. Results indicated that the AT-genome loci had significantly higher levels of diversity and lower levels of LD relative to homoelogous loci from the DT-genome. Additionally, all loci showed signatures of a population size expansion after a bottleneck or selective sweep and/or purifying selection. As previously shown for a sister tetraploid taxa (G. hirsutum), gene conversion resulting from a DT-genome allele invasion into the AT-genome likely explains the higher levels of diversity and lower levels of intragenic LD in the AT-genome. Given the relatively very low level of genetic diversity in elite lines, introduction of novel alleles from wild, land race, or obsolete lines into modern Pima cotton breeding programs is needed to expand the narrow gene pool of G. barbadense for continual yield improvements.

Anole lizards that inhabit the islands and mainland of the Caribbean basin have evolved morphological traits adapted to the microhabitat that they occupy. The anoles on these islands have been characterized as "ecomorphs" or morphologically and behaviorally-adapted groups, including: crown-giant, trunk-crown, trunk, grass-bush, twig, and trunk-ground. Ecomorphs display morphological features that are specifically adapted to the habitat that the anole occupies. One key morphological difference is tail length. While the anoles Anolis carolinensis and A. sagrei have similar ratios of tail length versus snout-to-vent length (SVL), they occupy different microhabitats. Specifically, A. carolinensis inhabits trunk-crown habitats while A. sagrei is found in trunk-ground regions. In this study, I focused on analysis of the caudal vertebrae of these two species, to determine if the structure of the osteological elements reflected differences in microhabitat adaptation. Skeletal preparations reveal that A. carolinensis have 40 \u2014 46 caudal vertebrae, and A. sagrei have 38 \u2014 49 caudal vertebrae. Transverse processes are present in Ca1-8 in A. carolinensis whereas transverse processes in A. sagrei span from Ca1-42 vertebrae. Ca6\u201440 have autotomy planes in A. sagrei, whereas only Ca8\u201417 have autotomy planes in A. carolinensis. These findings indicate that A. carolinensis are limited in the ability to autotomize their tail compared to A. sagrei. A. carolinensis, living higher in the trees than A. sagrei, might incur a greater impairment of locomotor function if autotomized. There appears to be no differences between males and females of both species in respect to vertebrae lengths. Differences between A. carolinensis and A. sagrei in terms of vertebral length are found in Ca12-15, 29-30, 34, and 37. The finding indicates that almost all caudal vertebrae between A. carolinensis and A. sagrei have similar relative lengths, but seven vertebrae have statistically significant differences. The biological significance of the findings is not clear, but functional and myological studies may help elucidate the reason of the observed differences.

The ringtail (Bassariscus astutus), a member of the Procyonidae, is capable of 180 degrees of hindlimb reversal during headfirst descent down a vertical substrate. The goal of this study was to determine the presence or absence of myological adaptations related to hindlimb reversal in the ringtail. Data for B. astutus are presented, including muscle weights and muscle maps ascertained from the dissection of four hindlimbs. Data from the current study were compared to published accounts of other species capable of hindlimb reversal, including procyonids (raccoon, coati, kinkajou, olingo), a mustelid (marten), palm civet, mongoose, tree squirrel, common tree shrew, and slow loris. Muscle mass data from this study demonstrate that the hip adductors of scansorial mammals are significantly more robust than those of terrestrial mammals, indicating a myological adaptation for climbing, but not necessarily hindlimb reversal. Among hindlimb reversers, the majority exhibit one belly of m. sartorius, the presence of m. extensor digiti I longus, and a fibular origin for m. fibularis longus. These characteristics indicate an emphasis on hip extension, ankle plantarflexion, and pes inversion. However, these characteristics are more likely due to phylogeny than hindlimb reversal because of their presence in closely-related non-reversers. Additional data on families outside of Carnivora may help determine if these myological traits are indeed due to phylogeny. Other myological data, such as moment arms and cross sectional areas, may provide evidence of adaptations for hindlimb reversal.

While a number of vertebrates, including fishes, salamanders, frogs, and lizards, display regenerative capacity, the process is not necessarily the same. It has been proposed that regeneration, while evolutionarily conserved, has diverged during evolution. However, the extent to which the mechanisms of regeneration have changed between taxa still remains elusive. In the salamander limb, cells dedifferentiate to a more plastic state and aggregate in the distal portion of the appendage to form a blastema, which is responsible for outgrowth and tissue development. In contrast, no such mechanism has been identified in lizards, and it is unclear to what extent evolutionary divergence between amniotes and anamniotes has altered this mechanism. Anolis carolinensis lizards are capable of regenerating their tails after stress-induced autotomy or self-amputation. In this investigation, the distribution of proliferating cells in early A. carolinensis tail regeneration was visualized by immunohistochemistry to examine the location and quantity of proliferating cells. An aggregate of proliferating cells at the distal region of the regenerate is considered indicative of blastema formation. Proliferating cell nuclear antigen (PCNA) and minichromosome maintenance complex component 2 (MCM2) were utilized as proliferation markers. Positive cells were counted for each tail (n=9, n=8 respectively). The percent of proliferating cells at the tip and base of the regenerating tail were compared with a one-way ANOVA statistical test. Both markers showed no significant difference (P=0.585, P=0.603 respectively) indicating absence of a blastema-like structure. These results suggest an alternative mechanism of regeneration in lizards and potentially other amniotes.

A literature review summarizing the current status of conservation efforts of the Mojave Desert tortoise (Gopherus agassizii) including a brief overview of the Endangered Species Act (ESA) and its applicability to this species' conservation. A genetic and physiological comparison of the morphologically similar Mojave species with the Sonoran (Gopherus morafkai) species proceeded by an analysis of if and how the ESA should apply to the Sonoran population. Analysis of current plans and interagency cooperations followed by a multi-step proposal on how best to conserve the Sonoran population of Desert tortoise.