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- All Subjects: Biology
- Genre: Doctoral Dissertation
- Creators: Arizona State University
- Resource Type: Text
Description
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.
ContributorsPalade, Joanna (Author) / Wilson-Rawls, Norma (Thesis advisor) / Rawls, Jeffrey (Committee member) / Kusumi, Kenro (Committee member) / Newbern, Jason (Committee member) / Stabenfeldt, Sarah (Committee member) / Arizona State University (Publisher)
Created2020
Description
Ant colonies provide numerous opportunities to study communication systems that maintain the cohesion of eusocial groups. In many ant species, workers have retained their ovaries and the ability to produce male offspring; however, they generally refrain from producing their own sons when a fertile queen is present in the colony. Although mechanisms that facilitate the communication of the presence of a fertile queen to all members of the colony have been highly studied, those studies have often overlooked the added challenge faced by polydomous species, which divide their nests across as many as one hundred satellite nests resulting in workers potentially having infrequent contact with the queen. In these polydomous contexts, regulatory phenotypes must extend beyond the immediate spatial influence of the queen.
This work investigates mechanisms that can extend the spatial reach of fertility signaling and reproductive regulation in three polydomous ant species. In Novomessor cockerelli, the presence of larvae but not eggs is shown to inhibit worker reproduction. Then, in Camponotus floridanus, 3-methylheptacosane found on the queen cuticle and queen-laid eggs is verified as a releaser pheromone sufficient to disrupt normally occurring aggressive behavior toward foreign workers. Finally, the volatile and cuticular hydrocarbon pheromones present on the cuticle of Oecophylla smaragdina queens are shown to release strong attraction response by workers; when coupled with previous work, this result suggests that these chemicals may underly both the formation of a worker retinue around the queen as well as egg-located mechanisms of reproductive regulation in distant satellite nests. Whereas most previous studies have focused on the short-range role of hydrocarbons on the cuticle of the queen, these studies demonstrate that eusocial insects may employ longer range regulatory mechanisms. Both queen volatiles and distributed brood can extend the range of queen fertility signaling, and the use of larvae for fertility signaling suggest that feeding itself may be a non-chemical mechanism for reproductive regulation. Although trail laying in mass-recruiting ants is often used as an example of complex communication, reproductive regulation in ants may be a similarly complex example of insect communication, especially in the case of large, polydomous ant colonies.
This work investigates mechanisms that can extend the spatial reach of fertility signaling and reproductive regulation in three polydomous ant species. In Novomessor cockerelli, the presence of larvae but not eggs is shown to inhibit worker reproduction. Then, in Camponotus floridanus, 3-methylheptacosane found on the queen cuticle and queen-laid eggs is verified as a releaser pheromone sufficient to disrupt normally occurring aggressive behavior toward foreign workers. Finally, the volatile and cuticular hydrocarbon pheromones present on the cuticle of Oecophylla smaragdina queens are shown to release strong attraction response by workers; when coupled with previous work, this result suggests that these chemicals may underly both the formation of a worker retinue around the queen as well as egg-located mechanisms of reproductive regulation in distant satellite nests. Whereas most previous studies have focused on the short-range role of hydrocarbons on the cuticle of the queen, these studies demonstrate that eusocial insects may employ longer range regulatory mechanisms. Both queen volatiles and distributed brood can extend the range of queen fertility signaling, and the use of larvae for fertility signaling suggest that feeding itself may be a non-chemical mechanism for reproductive regulation. Although trail laying in mass-recruiting ants is often used as an example of complex communication, reproductive regulation in ants may be a similarly complex example of insect communication, especially in the case of large, polydomous ant colonies.
ContributorsEbie, Jessica (Author) / Liebig, Jürgen (Thesis advisor) / Hölldobler, Bert (Thesis advisor) / Pratt, Stephen (Committee member) / Smith, Brian (Committee member) / Rutowski, Ronald (Committee member) / Arizona State University (Publisher)
Created2020
Description
Social insect groups, such as bees, termites, and ants, epitomize the emergence of group-level behaviors from the aggregated actions and interactions of individuals. Ants have the unique advantage that whole colonies can be observed in artificial, laboratory nests, and each individual's behavior can be continuously tracked using imaging software. In this dissertation, I study two group behaviors: (1) the spread of alarm signals from three agitated ants to a group of 61 quiescent nestmates, and (2) the reduction in per-capita energy use as colonies scale in size from tens of ants to thousands. For my first experiment, I track the motion of Pogonomyrmex californicus ants using an overhead camera, and I analyze how propagation of an initial alarm stimulus affects their walking speeds. I then build an agent-based model that simulates two-dimensional ant motion and the spread of the alarmed state. I find that implementing a simple set of rules for motion and alarm signal transmission reproduces the empirically observed speed dynamics. For the second experiment, I simulate social insect colony workers that collectively complete a set of tasks. By assuming that task switching is energetically costly, my model recovers a metabolic rate scaling pattern, known as hypometric metabolic scaling. This relationship, which predicts an organism's metabolic rate from its mass, is observed across a diverse set of social insect groups and animal species. The results suggest an explicit link between the degree of workers' task specialization and whole-colony energy use.
ContributorsLin, Michael Robert (Author) / Milner, Fabio A (Thesis advisor, Committee member) / Fewell, Jennifer H (Thesis advisor, Committee member) / Lampert, Adam (Committee member) / Arizona State University (Publisher)
Created2021
Description
The splicing of precursor messenger RNAs (pre-mRNAs) plays an essential role in dictating the mature mRNA profiles of eukaryotic cells. Mis-regulation of splicing, due to mutations in pre-mRNAs or in components of the splicing machinery, is associated with many diseases. Therefore, knowledge of pre-mRNA splicing mechanisms is required to understand gene expression regulation during states of homeostasis and disease, and for the development of therapeutic interventions.Splicing is catalyzed by the spliceosome, a dynamic and protein-rich ribozyme composed of five small nuclear ribonucleoproteins (snRNPs) and ~170 auxiliary factors. Early interactions that occur in prespliceosomal complexes formed by the 5′- and 3′-splice-site bound U1 and U2 snRNPs are responsible for committing introns for removal. However, the mechanisms underlying these early interactions remain to be fully characterized for understanding the influence of alternative splicing factors and the impact of recurrent disease-associated mutations in prespliceosomal proteins. The goal of my dissertation research was to delineate the role of the U1 small nuclear RNA (snRNA) during prespliceosome assembly. By applying a cellular minigene reporter assay and a variety of in vitro techniques including cell-free protein expression, UV-crosslinking, electrophoretic mobility shift assays, surface plasmon resonance, and RNA affinity purification, my work establishes critical roles for the U1 snRNA stem-loops 3 (SL3) and 4 (SL4) in formation of intron definition interactions during prespliceosome assembly. Previously, the SL4 of the U1 snRNA was shown to form a molecular bridge across introns by contacting the U2-specific splicing factor 3A1 (SF3A1). I identified the Ubiquitin-like domain of SF3A1 as a non-canonical RNA binding domain responsible for U1-SL4 binding. I also determined a role for the SL3 region of the U1 snRNA in splicing and characterized the spliceosomal RNA helicase UAP56 as an SL3 interacting protein. By knocking-down the SL3- and SL4-interacting proteins, I confirmed that U1 splicing activity in vivo relies on UAP56 and SF3A1 and that their functions are interdependent. These findings, in addition to the observations made using in vitro splicing assays, support a model whereby UAP56, through its interaction with U1-SL3, enhances the cross-intron interaction between U1-SL4 and SF3A1 to promote prespliceosome formation.
ContributorsMartelly, William (Author) / Sharma, Shalini (Thesis advisor) / Mangone, Marco (Thesis advisor) / Gustin, Kurt (Committee member) / Chen, Julian (Committee member) / Arizona State University (Publisher)
Created2021
Description
The flexibility and robustness of social insect colonies, when they cope with challenges as integrated units, raise many questions, such as how hundreds and thousands of individual local responses are coordinated without a central controlling process. Answering such questions requires: 1. Quantifiable collective responses of colonies under specific scenarios; 2. Decomposability of the collective colony-level response into individual responses; and 3. Mechanisms to integrate the colony- and individual-level responses. In the first part of my dissertation, I explore coordinated collective responses of colonies in during the alarm response to an alarmed nestmate (chapter 2&3). I develop a machine-learning approach to quantitatively estimate the collective and individual alarm response (chapter 2). Using this methodology, I demonstrate that colony alarm responses to the introduction of alarmed nestmates can be decomposed into immediately cascading, followed by variable dampening processes. Each of those processes are found to be modulated by variation in individual alarm responsiveness, as measured by alarm response threshold and persistence of alarm behavior. This variation is modulated in turn by environmental context, in particular with task-related social context (chapter 3). In the second part of my dissertation, I examine the mechanisms responsible for colonial changes in metabolic rate during ontogeny. Prior studies have found that larger ant colonies (as for larger organisms) have lower mass-specific metabolic rates, but the mechanisms remain unclear. In a 3.5-year study on 25 colonies, metabolic rates of colonies and colony components were measured during ontogeny (chapter 4). The scaling of metabolic rate during ontogeny was fit better by segmented regression or quadratic regression models than simple linear regression models, showing that colonies do not follow a universal power-law of metabolism during the ontogenetic development. Furthermore, I showed that the scaling of colonial metabolic rates can be primarily explained by changes in the ratio of brood to adult workers, which nonlinearly affects colonial metabolic rates. At high ratios of brood to workers, colony metabolic rates are low because the metabolic rate of larvae and pupae are much lower than adult workers. However, the high colony metabolic rates were observed in colonies with moderate brood: adult ratios, because higher ratios cause adult workers to be more active and have higher metabolic rates, presumably due to the extra work required to feed more brood.
ContributorsGuo, Xiaohui (Author) / Fewell, Jennifer H (Thesis advisor) / Kang, Yun (Thesis advisor) / Harrison, Jon F (Committee member) / Liebig, Juergen (Committee member) / Pratt, Stephen C (Committee member) / Pavlic, Theodore P (Committee member) / Arizona State University (Publisher)
Created2021
Description
The control, function, and evolution of sleep in animals has received little attention compared to many other fitness-relevant animal behaviors. Though natural selection has largely been thought of as the driving evolutionary force shaping sleep biology, sexual and social selection may also have transformative effects on sleep quantity and quality in animals. An overarching hypothesis is that increased levels of investment into inter-sexual choice and intra-sexual competition will reduce sleep. An alternative hypothesis is that sexual ornamentation (e.g. avian plumage coloration and song) may have evolved to communicate sleep health and may therefore be positively related to sleep investment. In this dissertation, I studied how sleep is related to components of sexual and social selection in animals (mostly in birds). I first reviewed the literature for empirical examples of how social and sexual selection drive animal sleep patterns and found support for this relationship in some common types of inter-individual interactions (e.g. mating, intra-sexual competition, parent-offspring interactions, group interactions); I also provided new ideas and hypotheses for future research. I then tested associations between sleep behavior with expression of ornaments (song and plumage coloration), using the house finch (Haemorhous mexicanus) as a model system. For both color and song, I found support for the hypothesis that individuals with exaggerated ornaments slept deeper and longer, suggesting that sleep is a critical resource for ornament elaboration and/or may be communicated by both types of sexual signal. Following this, I tested the phylogenetic association between sleep and social/sexual selection as well as other life-history traits across birds. I found that more territorial bird species sleep less, that polygynous birds sleep more than monogamous and polygynandrous birds, and that birds migrating longer distances sleep less and have less REM sleep. Finally, in the interest of applying basic knowledge about sleep biology to current global problems, I found support for the hypothesis that house finches from city environments have developed resilience to artificial light pollution at night. Altogether, I found that social, sexual, and life-history traits are indeed important and overlooked drivers of sleep behavior from multiple levels of analysis.
ContributorsHutton, Pierce (Author) / McGraw, Kevin J (Thesis advisor) / Rutowski, Ronald L (Committee member) / Deviche, Pierre J (Committee member) / Sweazea, Karen L (Committee member) / Lesku, John A (Committee member) / Arizona State University (Publisher)
Created2021
Description
The living world is replete with easily observed structural adaptations (e.g. teeth, claws, and stingers), but behavioral adaptations are no less impressive. Conspecific aggression can be defined as any harmful action directed by one animal at another of the same species. Because it is a potentially risky and costly behavior, aggression should be elicited only under optimal conditions. In honeybees, nestmate recognition is considered the driving factor determining whether colony guards will aggress against other honeybees attempting to gain entry to the colony. Models and empirical research support the conclusion that nestmate recognition should be favored over direct kin recognition. Thus, bees tend to use environmentally mediated cues associated with their colonies (e.g. colony odors) to recognize nestmates. The framework of nestmate recognition suggests that non-nestmates should always be aggressed against while nestmates should always be accepted. However, aggression towards nestmates and acceptance of non-nestmates are seen in a wide variety of eusocial insects, including honeybees. These are typically classified as rejection errors and acceptance errors, respectively. As such, they can be explained using signal detection theory and optimal acceptance threshold models, which postulate that recognition errors are inevitable if there is overlap in the cues used to distinguish “desirables” (fitness-enhancing) from “undesirables” (fitness-decrementing) conspecifics. In the context of social insects desirables are presumed to be nestmates and undesirables are presumed to be non-nestmates. I propose that honeybees may make more refined decisions concerning what conspecifics are desirable and undesirable, accounting for at least some of the phenomena previously reported as recognition errors. Some “errors” may be the result of guard bees responding to cues associated with threats and benefits beyond nestmate identity. I show that less threatening neighbors receive less aggression than highly threatening strangers. I show that well-fed colonies exhibit less aggression and that bees from well-fed colonies receive less aggression. I provide evidence that honeybees may decrease aggression towards nestmates and non-nestmate not involved in robbing while increasing aggression towards non-nestmate from a robber colony. Lastly, I show that pollen bearing foragers, regardless of nestmate identity, receive little to no aggression compared to non-pollen bearing foragers.
ContributorsJackson, Jonathan Cole (Author) / Pratt, Stephen (Thesis advisor) / Rutowski, Ronald (Committee member) / Fewell, Jennifer (Committee member) / Amazeen, Nia (Committee member) / Kaftanoglu, Osman (Committee member) / Arizona State University (Publisher)
Created2021
Description
Lipolysis or hydrolysis of triglyceride (TG) stored within intracellular lipid droplets (LD), is vital to maintaining metabolic homeostasis in mammals. Regulation of lipolysis and subsequent utilization of liberated fatty acids impacts cellular and organismal functions including body fat accumulation and thermogenesis. Adipose triglyceride lipase (ATGL) is the intracellular rate-limiting enzyme responsible for catalyzing hydrolysis of TG to diacylglycerol (DAG), the initial step of the lipolytic reaction. G0/G1 switch gene-2 (G0S2) and hypoxia-inducible gene-2 (HIG2) are selective inhibitors of ATGL. G0S2 facilitates accumulation of TG in the liver and adipose tissue, while HIG2 functions under hypoxic conditions. Sequence analysis and mutagenesis were used to confirm the presence of conserved domains between these proteins, and that these domains are required for efficient binding and inhibition of ATGL. Further analysis revealed a Positive sequence (Pos-Seq)-LD binding motif in G0S2 but not HIG2. The Pos-Seq mediated ATGL-independent localization to LD and was required for achieving maximal inhibition of ATGL activity by G0S2. Identification and mutational analysis of this motif revealed distinct mechanisms for HIG2 and G0S2 LD association. In addition to molecular characterization of known protein inhibitors of lipolysis, an intracellular member of the apolipoprotein L (ApoL) family, ApoL6, was also identified as a LD and mitochondria associated protein expressed in adipose tissue. Brown adipose tissue uses fatty acids as fuel for increasing its energy output as heat during acute responses to cold exposure. A Comprehensive Lab Animal Monitoring System was used to compare heat production at room temperature (RT) and 4oC in transgenic animals overexpressing ApoL6 in brown adipose tissue. Overexpression of ApoL6 delayed utilization of long-chain fatty acids (LCFAs) as a fuel source while promoting an enhanced thermogenic response during initial cold exposure. ApoL6 mediated inhibition of LCFA utilization results from binding of ApoL6 to Mitochondrial Trifunctional Protein (MTP/TFP), which catalyzes mitochondrial β-oxidation. Indirect calorimetry and fasting acute cold exposure experiments suggest the augmented thermogenic profile of ApoL6 transgenic animals is a result of enhanced utilization of medium-chain fatty acids (MCFAs), glucose, and amino acids as fuel sources. Cumulatively these results indicate multiple mechanisms for regulation lipolysis and fatty acid utilization.
ContributorsCampbell, Latoya E (Author) / Lake, Douglas (Thesis advisor) / Liu, Jun (Committee member) / Folmes, Clifford (Committee member) / Sweazea, Karen (Committee member) / Baluch, Debra (Committee member) / Arizona State University (Publisher)
Created2021
Description
This research investigates how two potential sentinel species (the Galápagos Sea Lion (Zalophus wollebaeki) and the Guiana Dolphin (Sotalia guianensis)) respond to environmental factors, at both the large-scale and fine-scale levels. Sentinel species, defined as organisms able to respond to ecosystem variability and/or change in a timely and measurable way to nowcast or forecast otherwise unobserved environmental changes, can help mitigate or even avoid changes deleterious to both wildlife and human communities. Using two long-term datasets and a suite of respective social metrics and environmental factors, I analyzed potential external influences on these two species’ behavioral ecology. My overall findings suggest that apex marine mammals respond differently to their surroundings at large-scale vs. fine-scale, and highlight the importance of including a range of environmental factors that include anthropogenic effects. Galápagos Sea Lions specifically respond to thermoregulation-linked factors, such as substrate temperature, and anthropogenic factors such as human presence and activity type. Guiana Dolphin social metrics are significantly related with traits linked to environmental water quality, water transparency. I expand on the sentinel implications of these results and introduce sample methodology and results for sentinel species based on the Guiana Dolphin case study.
ContributorsAbalo, Iroko Akoua Enyo (Author) / Pratt, Stephen S (Thesis advisor) / Polidoro, Beth B (Thesis advisor) / Abbott, Joshua J (Committee member) / Ferry, Lara L (Committee member) / Arizona State University (Publisher)
Created2021
Description
This dissertation investigates how ideas of the right relationships among science, the public, and collective decision-making about science and technology come to be envisioned in constructions of public engagement. In particular, it explores how public engagement has come to be constructed in discourse around gene editing to better understand how it holds together with visions for good, democratic governance of those technologies and with what effects. Using a conceptual idiom of the co-production of science and the social order, I investigate the mutual formation of scientific expertise, responsibility, and democracy through constructions of public engagement. I begin by tracing dominant historical narratives of contemporary public engagement as a continuation of public understanding of science’s projects of social ordering for democratic society. I then analyze collections of prominent expert meetings, publications, discussions, and interventions about development, governance, and societal implications human heritable germline gene editing and gene drives that developed in tandem with commitments to public engagement around those technologies. Synthesizing the evidence from across gene editing discourse, I offer a constructive critique of constructions of public engagement as expressions and evidence of scientific responsibility as ultimately reasserting and reinforcing of scientific experts' authority in gene editing decision-making, despite intentions for public engagement to extend decision-making participation and power to publics. Such constructions of public engagement go unrecognized in gene editing discourse and thereby subtly reinforce broader visions of scientific expertise as essential to good governance by underwriting the legitimacy and authority of scientific experts to act on behalf of public interests. I further argue that the reinforcement of scientific expert authority in gene editing discourse through public engagement also centers scientific experts in a sociotechnical imaginary that I call “not for science alone.” This sociotechnical imaginary envisions scientific experts as guardians and guarantors of good, democratic governance. I then propose a possible alternatives to public engagement alone to improve gene editing governance by orienting discourse around notions of public accountability for potential shared benefits and collective harms of gene editing.
ContributorsRoss, Christian (Author) / Hurlbut, James B. (Thesis advisor) / Maienschein, Jane (Thesis advisor) / Collins, James P. (Committee member) / Crow, Michael M. (Committee member) / Sarewitz, Daniel R. (Committee member) / Arizona State University (Publisher)
Created2021