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Description
Induced pluripotent stem cells (iPSCs) are an intriguing approach for neurological disease modeling, because neural lineage-specific cell types that retain the donors' complex genetics can be established in vitro. The statistical power of these iPSC-based models, however, is dependent on accurate diagnoses of the somatic cell donors; unfortunately, many neurodegenerative

Induced pluripotent stem cells (iPSCs) are an intriguing approach for neurological disease modeling, because neural lineage-specific cell types that retain the donors' complex genetics can be established in vitro. The statistical power of these iPSC-based models, however, is dependent on accurate diagnoses of the somatic cell donors; unfortunately, many neurodegenerative diseases are commonly misdiagnosed in live human subjects. Postmortem histopathological examination of a donor's brain, combined with premortem clinical criteria, is often the most robust approach to correctly classify an individual as a disease-specific case or unaffected control. We describe the establishment of primary dermal fibroblasts cells lines from 28 autopsy donors. These fibroblasts were used to examine the proliferative effects of establishment protocol, tissue amount, biopsy site, and donor age. As proof-of-principle, iPSCs were generated from fibroblasts from a 75-year-old male, whole body donor, defined as an unaffected neurological control by both clinical and histopathological criteria. To our knowledge, this is the first study describing autopsy donor-derived somatic cells being used for iPSC generation and subsequent neural differentiation. This unique approach also enables us to compare iPSC-derived cell cultures to endogenous tissues from the same donor. We utilized RNA sequencing (RNA-Seq) to evaluate the transcriptional progression of in vitro-differentiated neural cells (over a timecourse of 0, 35, 70, 105 and 140 days), and compared this with donor-identical temporal lobe tissue. We observed in vitro progression towards the reference brain tissue, supported by (i) a significant increasing monotonic correlation between the days of our timecourse and the number of actively transcribed protein-coding genes and long intergenic non-coding RNAs (lincRNAs) (P < 0.05), consistent with the transcriptional complexity of the brain, (ii) an increase in CpG methylation after neural differentiation that resembled the epigenomic signature of the endogenous tissue, and (iii) a significant decreasing monotonic correlation between the days of our timecourse and the percent of in vitro to brain-tissue differences (P < 0.05) for tissue-specific protein-coding genes and all putative lincRNAs. These studies support the utility of autopsy donors' somatic cells for iPSC-based neurological disease models, and provide evidence that in vitro neural differentiation can result in physiologically progression.
ContributorsHjelm, Brooke E (Author) / Craig, David W. (Thesis advisor) / Wilson-Rawls, Norma J. (Thesis advisor) / Huentelman, Matthew J. (Committee member) / Mason, Hugh S. (Committee member) / Kusumi, Kenro (Committee member) / Arizona State University (Publisher)
Created2013
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Description
The lack of substantive, multi-dimensional perspectives on civic space planning and design has undermined the potential role of these valuable social and ecological amenities in advancing urban sustainability goals. Responding to these deficiencies, this dissertation utilized mixed quantitative and qualitative methods and synthesized multiple social and natural science perspectives to

The lack of substantive, multi-dimensional perspectives on civic space planning and design has undermined the potential role of these valuable social and ecological amenities in advancing urban sustainability goals. Responding to these deficiencies, this dissertation utilized mixed quantitative and qualitative methods and synthesized multiple social and natural science perspectives to inform the development of progressive civic space planning and design, theory, and public policy aimed at improving the social, economic, and environmental health of cities. Using Phoenix, Arizona as a case study, the analysis was tailored to arid cities, yet the products and findings are flexible enough to be geographically customized to the social, environmental, built, and public policy goals of other urbanized regions. Organized into three articles, the first paper applies geospatial and statistical methods to analyze and classify urban parks in Phoenix based on multiple social, ecological, and built criteria, including landuse-land cover, `greenness,' and site amenities, as well as the socio- economic and built characteristics of park neighborhoods. The second article uses spatial empirical analysis to rezone the City of Phoenix following transect form-based code. The current park system was then assessed within this framework and recommendations are presented to inform the planning and design of civic spaces sensitive to their social and built context. The final paper culminates in the development of a planning tool and site design guidelines for civic space planning and design across the urban-to-natural gradient augmented with multiple ecosystem service considerations and tailored to desert cities.
ContributorsIbes, Dorothy (Author) / Talen, Emily (Thesis advisor) / Boone, Christopher (Committee member) / Crewe, Katherine (Committee member) / Arizona State University (Publisher)
Created2013
Description
Well-established model systems exist in four out of the seven major classes of vertebrates. These include the mouse, chicken, frog and zebrafish. Noticeably missing from this list is a reptilian model organism for comparative studies between the vertebrates and for studies of biological processes unique to reptiles. To help fill

Well-established model systems exist in four out of the seven major classes of vertebrates. These include the mouse, chicken, frog and zebrafish. Noticeably missing from this list is a reptilian model organism for comparative studies between the vertebrates and for studies of biological processes unique to reptiles. To help fill in this gap the green anole lizard, Anolis carolinensis, is being adapted as a model organism. Despite the recent release of the complete genomic sequence of the A. carolinensis, the lizard lacks some resources to aid researchers in their studies. Particularly, the lack of transcriptomic resources for lizard has made it difficult to identify genes complete with alternative splice forms and untranslated regions (UTRs). As part of this work the genome annotation for A. carolinensis was improved through next generation sequencing and assembly of the transcriptomes from 14 different adult and embryonic tissues. This revised annotation of the lizard will improve comparative studies between vertebrates, as well as studies within A. carolinensis itself, by providing more accurate gene models, which provide the bases for molecular studies. To demonstrate the utility of the improved annotations and reptilian model organism, the developmental process of somitogenesis in the lizard was analyzed and compared with other vertebrates. This study identified several key features both divergent and convergent between the vertebrates, which was not previously known before analysis of a reptilian model organism. The improved genome annotations have also allowed for molecular studies of tail regeneration in the lizard. With the annotation of 3' UTR sequences and next generation sequencing, it is now possible to do expressional studies of miRNA and predict their mRNA target transcripts at genomic scale. Through next generation small RNA sequencing and subsequent analysis, several differentially expressed miRNAs were identified in the regenerating tail, suggesting miRNA may play a key role in regulating this process in lizards. Through miRNA target prediction several key biological pathways were identified as potentially under the regulation of miRNAs during tail regeneration. In total, this work has both helped advance A. carolinensis as model system and displayed the utility of a reptilian model system.
ContributorsEckalbar, Walter L (Author) / Kusumi, Kenro (Thesis advisor) / Huentelman, Matthew (Committee member) / Rawls, Jeffery (Committee member) / Wilson-Rawls, Norma (Committee member) / Arizona State University (Publisher)
Created2012
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Description
The explicit role of soil organisms in shaping soil health, rates of pedogenesis, and resistance to erosion has only just recently begun to be explored in the last century. However, much of the research regarding soil biota and soil processes is centered on maintaining soil fertility (e.g., plant nutrient availability)

The explicit role of soil organisms in shaping soil health, rates of pedogenesis, and resistance to erosion has only just recently begun to be explored in the last century. However, much of the research regarding soil biota and soil processes is centered on maintaining soil fertility (e.g., plant nutrient availability) and soil structure in mesic- and agro- ecosystems. Despite the empirical and theoretical strides made in soil ecology over the last few decades, questions regarding ecosystem function and soil processes remain, especially for arid areas. Arid areas have unique ecosystem biogeochemistry, decomposition processes, and soil microbial responses to moisture inputs that deviate from predictions derived using data generated in more mesic systems. For example, current paradigm predicts that soil microbes will respond positively to increasing moisture inputs in a water-limited environment, yet data collected in arid regions are not congruent with this hypothesis. The influence of abiotic factors on litter decomposition rates (e.g., photodegradation), litter quality and availability, soil moisture pulse size, and resulting feedbacks on detrital food web structure must be explicitly considered for advancing our understanding of arid land ecology. However, empirical data coupling arid belowground food webs and ecosystem processes are lacking. My dissertation explores the resource controls (soil organic matter and soil moisture) on food web network structure, size, and presence/absence of expected belowground trophic groups across a variety of sites in Arizona.
ContributorsWyant, Karl Arthur (Author) / Sabo, John L (Thesis advisor) / Elser, James J (Committee member) / Childers, Daniel L. (Committee member) / Hall, Sharon J (Committee member) / Stromberg, Juliet C. (Committee member) / Arizona State University (Publisher)
Created2014
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Description
Like individual organisms, complex social groups are able to maintain predictable trajectories of growth, from initial colony foundation to mature reproductively capable units. They do so while simultaneously responding flexibly to variation in nutrient availability and intake. Leafcutter ant colonies function as tri-trophic systems, in which the ants harvest vegetation

Like individual organisms, complex social groups are able to maintain predictable trajectories of growth, from initial colony foundation to mature reproductively capable units. They do so while simultaneously responding flexibly to variation in nutrient availability and intake. Leafcutter ant colonies function as tri-trophic systems, in which the ants harvest vegetation to grow a fungus that, in turn, serves as food for the colony. Fungal growth rates and colony worker production are interdependent, regulated by nutritional and behavioral feedbacks. Fungal growth and quality are directly affected by worker foraging decisions, while worker production is, in turn, dependent on the amount and condition of the fungus. In this dissertation, I first characterized the growth relationship between the workers and the fungus of the desert leafcutter ant Acromyrmex versicolor during early stages of colony development, from colony foundation by groups of queens through the beginnings of exponential growth. I found that this relationship undergoes a period of slow growth and instability when workers first emerge, and then becomes allometrically positive. I then evaluated how mass and element ratios of resources collected by the ants are translated into fungus and worker population growth, and refuse, finding that colony digestive efficiency is comparable to digestive efficiencies of other herbivorous insects and ruminants. To test how colonies behaviorally respond to perturbations of the fungus garden, I quantified activity levels and task performance of workers in colonies with either supplemented or diminished fungus gardens, and found that colonies adjusted activity and task allocation in response to the fungus garden size. Finally, to identify possible forms of nutrient limitation, I measured how colony performance was affected by changes in the relative amounts of carbohydrates, protein, and phosphorus available in the resources used to grow the fungus garden. From this experiment, I concluded that colony growth is primarily carbohydrate-limited.
ContributorsClark, Rebecca, 1981- (Author) / Fewell, Jennifer H (Thesis advisor) / Mueller, Ulrich (Committee member) / Liebig, Juergen (Committee member) / Elser, James (Committee member) / Harrison, Jon (Committee member) / Arizona State University (Publisher)
Created2011
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Description
Damage to the central nervous system due to spinal cord or traumatic brain injury, as well as degenerative musculoskeletal disorders such as arthritis, drastically impact the quality of life. Regeneration of complex structures is quite limited in mammals, though other vertebrates possess this ability. Lizards are the most closely related

Damage to the central nervous system due to spinal cord or traumatic brain injury, as well as degenerative musculoskeletal disorders such as arthritis, drastically impact the quality of life. Regeneration of complex structures is quite limited in mammals, though other vertebrates possess this ability. Lizards are the most closely related organism to humans that can regenerate de novo skeletal muscle, hyaline cartilage, spinal cord, vasculature, and skin. Progress in studying the cellular and molecular mechanisms of lizard regeneration has previously been limited by a lack of genomic resources. Building on the release of the genome of the green anole, Anolis carolinensis, we developed a second generation, robust RNA-Seq-based genome annotation, and performed the first transcriptomic analysis of tail regeneration in this species. In order to investigate gene expression in regenerating tissue, we performed whole transcriptome and microRNA transcriptome analysis of regenerating tail tip and base and associated tissues, identifying key genetic targets in the regenerative process. These studies have identified components of a genetic program for regeneration in the lizard that includes both developmental and adult repair mechanisms shared with mammals, indicating value in the translation of these findings to future regenerative therapies.
ContributorsHutchins, Elizabeth (Author) / Kusumi, Kenro (Thesis advisor) / Rawls, Jeffrey A. (Committee member) / Denardo, Dale F. (Committee member) / Huentelman, Matthew J. (Committee member) / Arizona State University (Publisher)
Created2015
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Description

River and riparian areas are important foraging habitat for insectivorous bats. Numerous studies have shown that aquatic insects provide an important trophic resource to terrestrial consumers, including bats, and are key in regulating population size and species interactions in terrestrial food webs. Yet these studies have generally ignored how structural

River and riparian areas are important foraging habitat for insectivorous bats. Numerous studies have shown that aquatic insects provide an important trophic resource to terrestrial consumers, including bats, and are key in regulating population size and species interactions in terrestrial food webs. Yet these studies have generally ignored how structural characteristics of the riverine landscape influence trophic resource availability or how terrestrial consumers respond to ensuing spatial and temporal patterns of trophic resources. Moreover, few studies have examined linkages between a stream's hydrologic regime and the timing and magnitude of aquatic insect availability. The main objective of my dissertation is to understand the causes of bat distributions in space and time. Specifically, I examine how trophic resource availability, structural components of riverine landscapes (channel confinement and riparian vegetation structure), and hydrologic regimes (flow permanence and timing of floods) mediate spatial and temporal patterns in bat activity. First, I show that river channel confinement determines bat activity along a river's longitudinal axis (directly above the river), while trophic resources appear to have stronger effects across a river's lateral (with distance from the river) axis. Second, I show that flow intermittency affects bat foraging activity indirectly via its effects on trophic resource availability. Seasonal river drying appears to have complex effects on bat foraging activity, initially causing imperfect tracking by consumers of localized concentrations of resources but later resulting in disappearance of both insects and bats after complete river drying. Third, I show that resource tracking by bats varies among streams with contrasting patterns of trophic resource availability and this variation appears to be in response to differences in the timing of aquatic insect emergence, duration and magnitude of emergence, and adult body size of emergent aquatic insects. Finally, I show that aquatic insects directly influence bat activity along a desert stream and that riparian vegetation composition affects bat activity, but only indirectly, via effects on aquatic insect availability. Overall, my results show river channel confinement, riparian vegetation structure, flow permanence, and the timing of floods influence spatial and temporal patterns in bat distributions; but these effects are indirect by influencing the ability of bats to track trophic resources in space and time.

ContributorsHagen, Elizabeth M (Author) / Sabo, John L (Thesis advisor) / Fisher, Stuart G. (Committee member) / Grimm, Nancy (Committee member) / Schmeeckle, Mark W (Committee member) / Stromberg, Juliet C. (Committee member) / Arizona State University (Publisher)
Created2010
Description

Agassiz’s desert tortoise (Gopherus agassizii) is a long-lived species native to the Mojave Desert and is listed as threatened under the US Endangered Species Act. To aid conservation efforts for preserving the genetic diversity of this species, we generated a whole genome reference sequence with an annotation based on dee

Agassiz’s desert tortoise (Gopherus agassizii) is a long-lived species native to the Mojave Desert and is listed as threatened under the US Endangered Species Act. To aid conservation efforts for preserving the genetic diversity of this species, we generated a whole genome reference sequence with an annotation based on deep transcriptome sequences of adult skeletal muscle, lung, brain, and blood. The draft genome assembly for G. agassizii has a scaffold N50 length of 252 kbp and a total length of 2.4 Gbp. Genome annotation reveals 20,172 protein-coding genes in the G. agassizii assembly, and that gene structure is more similar to chicken than other turtles. We provide a series of comparative analyses demonstrating (1) that turtles are among the slowest-evolving genome-enabled reptiles, (2) amino acid changes in genes controlling desert tortoise traits such as shell development, longevity and osmoregulation, and (3) fixed variants across the Gopherus species complex in genes related to desert adaptations, including circadian rhythm and innate immune response. This G. agassizii genome reference and annotation is the first such resource for any tortoise, and will serve as a foundation for future analysis of the genetic basis of adaptations to the desert environment, allow for investigation into genomic factors affecting tortoise health, disease and longevity, and serve as a valuable resource for additional studies in this species complex.

Data Availability: All genomic and transcriptomic sequence files are available from the NIH-NCBI BioProject database (accession numbers PRJNA352725, PRJNA352726, and PRJNA281763). All genome assembly, transcriptome assembly, predicted protein, transcript, genome annotation, repeatmasker, phylogenetic trees, .vcf and GO enrichment files are available on Harvard Dataverse (doi:10.7910/DVN/EH2S9K).

ContributorsTollis, Marc (Author) / DeNardo, Dale F (Author) / Cornelius, John A (Author) / Dolby, Greer A (Author) / Edwards, Taylor (Author) / Henen, Brian T. (Author) / Karl, Alice E. (Author) / Murphy, Robert W. (Author) / Kusumi, Kenro (Author)
Created2017-05-31
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Description
Embryonic and juvenile development consist of a series of complex and rapid changes driven by a suite of crucially timed developmental cues within the cell. The developmental process begins at the moment of zygote activation, “jump-started” by maternal factors such as mRNA and proteins until transcription can be zygotically-driven. Regulation

Embryonic and juvenile development consist of a series of complex and rapid changes driven by a suite of crucially timed developmental cues within the cell. The developmental process begins at the moment of zygote activation, “jump-started” by maternal factors such as mRNA and proteins until transcription can be zygotically-driven. Regulation of transcription initiation plays a crucial role in this process, as minute changes in the timing, density, and characteristics of gene expression can have drastic effects on the zygote’s development. Specific promoter elements can be linked to different patterns of transcription, driving both ubiquitous and sharply regulated gene expression, thus forming the basis for the time-sensitive developmental processes. In order to better understand the genes expressed during different stages of development and the impact of promoter elements on transcription patterns and transcript concentrations within the cell, I created a Gene Expression and Promoter Atlas in two species within the cryptic species complex, Daphnia pulex. I surveyed five embryonic and two juvenile developmental stages in both a North American and mitochondrially European Daphnia pulex utilizing developmental landmarks to visually stages embryos. A total of 17,993 genes were identified in the European species and 15,295 were identified in the North American species, with 11,551 orthologs identified between the two. I utilized the transcription start site (TSS) profiling method STRIPE-seq to identify promoter motifs and RNA-seq to survey mRNA concentration at each stage, generating a wealth of genetic data. The methodology for library construction and the dataset generated therein provide an informative basis for further comparative developmental studies and the elucidation of full gene functionality in an emerging model organism.
ContributorsWalls, Sarah (Author) / Lynch, Michael (Thesis advisor) / Raborn, R. Taylor (Committee member) / Mangoni, Marco (Committee member) / Harris, Robin (Committee member) / Arizona State University (Publisher)
Created2022
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Description
Understanding the effects of fire on the Sonoran Desert is of critical importance as rising temperatures and changing weather patterns increase the frequency and size of wildfires. Seed banks are an important component in post-fire landscape recovery as the seeds that remain in the soil are an indicator of a

Understanding the effects of fire on the Sonoran Desert is of critical importance as rising temperatures and changing weather patterns increase the frequency and size of wildfires. Seed banks are an important component in post-fire landscape recovery as the seeds that remain in the soil are an indicator of a landscape’s future trajectory. The purpose of this study is to determine the lasting impacts of fire on the soil seed bank of the Sonoran Desert and to identify potential concerns affecting post-fire recovery and restoration. The study site was located in the Arizona Upland division of the Sonoran Desert, Arizona, United States. Soil samples were collected from five burned sites with increasing time since fire, and five nearby unburned sites used as a control. A seedling emergence test was conducted to investigate the density and richness of the seed bank of burned and unburned sites. Seed densities and species richness for sites were calculated using germination results. Findings were analyzed using non-parametric analyses comparing changes in burned and unburned sites over time. Results found that burn status and time since fire had no significant impact on seed density. Graminoid and forb densities were statistically consistent across burn status and time since fire. While species richness was consistent across both plot types, burned samples typically had fewer species than unburned samples. Burned and unburned plots revealed a dominance of annual species with limited presence of woody perennials. While seed densities and species richness are relatively unchanged across burned and unburned sites over time, the lack of woody perennials in the seed bank raises concerns about landscape recovery trajectories in burned sites. These results suggest that restoration efforts focused on maintaining the presence of woody trees and shrubs in the landscape may have the most impact.
ContributorsCountryman, Kristen (Author) / Makings, Elizabeth (Thesis advisor) / Pigg, Kathleen (Thesis advisor) / Lata, Mary (Committee member) / Arizona State University (Publisher)
Created2023