Animal Experimentation and Research Commons^™

Importance Of The Microhabitat And Microclimate Conditions In The Northern Gray-Cheeked Salamander (Plethodon Montanus) Across An Elevation Gradient, Trevor Chapman

Electronic Theses and Dissertations

The southern Appalachian Mountains have among the highest salamander diversity in the world, largely due to local speciation in the family Plethodontidae. Plethodontid salamanders (i.e., lungless salamanders) are particularly sensitive to habitat climate conditions due to their reliance on cutaneous respiration, and their immediate environmental conditions (microhabitat) likely influence their dispersion and activity more than the large-scale atmospheric conditions. The Northern Gray-cheeked salamander (Plethodon montanus) is restricted to high elevations in the Appalachian Mountains. Our goal was to investigate the relationship between P. montanus and its microhabitat by examining behavioral preference for climatic conditions, characterizing the microclimate with …

Internal Vertebral Morphology Of Bony Fishes Matches The Mechanical Demands Of Different Environments, Dana Baxter, Karly E. Cohen, Cassandra M. Donatelli, Eric D. Tytell

Engineering Faculty Articles and Research

Fishes have repeatedly evolved characteristic body shapes depending on how close they live to the substrate. Pelagic fishes live in open water and typically have narrow, streamlined body shapes; benthic and demersal fishes live close to the substrate; and demersal fishes often have deeper bodies. These shape differences are often associated with behavioral differences: pelagic fishes swim nearly constantly, demersal fishes tend to maneuver near the substrate, and benthic fishes often lie in wait on the substrate. We hypothesized that these morphological and behavioral differences would be reflected in the mechanical properties of the body, and specifically in vertebral column …

The Role Of The Hypoxia-Inducible Factor 2 In Pancreatic Cancer: Mechanisms Of Tumor Immunosuppression And Intestinal Radioprotection, Carolina Garcia Garcia

Dissertations and Theses (Open Access)

Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with dismal prognosis. The only curative option for patients is surgery, but over 80% of patients are not surgical candidates. Unfortunately, PDAC is resistant to the three remaining options. PDAC is characterized by a profoundly hypoxic and immunosuppressive stroma, which contributes to its therapeutic recalcitrance. Alpha-smooth muscle actin+ (αSMA+) cancer-associated fibroblasts (CAFs) are the most abundant stromal component, as well as mediators of stromal deposition. The hypoxia-inducible factors (HIF1 and HIF2) coordinate responses to hypoxia, yet, despite their known association to poor patient outcomes, their functions within the PDAC tumor microenvironment (TME) …

Maternal Responses In The Face Of Infection Risk, Patricia C. Lopes, Brenna M. G. Gormally, Aubrey Emmi, Delilah Schuerman, Chathuni Liyanage, Ursula K. Beattie, L. Michael Romero

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

When animals are sick, their physiology and behavior change in ways that can impact their offspring. Research is emerging showing that infection risk alone can also modify the physiology and behavior of healthy animals. If physiological responses to environments with high infection risk take place during reproduction, it is possible that they lead to maternal effects. Understanding whether and how high infection risk triggers maternal effects is important to elucidate how the impacts of infectious agents extend beyond infected individuals and how, in this way, they are even stronger evolutionary forces than already considered. Here, to evaluate the effects of …

Effects Of Chronic Stress On Safety Processing And Physiology In The Medial Prefrontal-Amygdala-Basal Forebrain Circuit, Itamar S. Grunfeld

Dissertations, Theses, and Capstone Projects

Chronic stress increases generalization of fear to non-threatening cues, a key symptom in numerous psychiatric mood disorders. However, the mechanisms through which stress impacts safety learning remain poorly understood. To probe the relationship between stress and safety learning, this dissertation employed multiple behaviors, in conjunction with in-vivo multisite physiology during explicit safety and fear discrimination learning. In Chapter 1, I outline the role of chronic stress in driving neurological adaptations that result in generalized fear and highlight how this occurs because of impaired safety cue encoding. In Chapter 2, I show that chronic stress in the form of social defeat …

Developing A Novel Place Preference Assay To Compare Drosophila Species Over Time, Martha M. Brinson

Honors Theses

Across phylogeny, integration of external factors, memory, and internal states of the organism dictate organismal behavior and mechanisms. The underlying genetic components can affect these behaviors such as in genomic changes arising from speciation. In this thesis, a new place preference assay was evaluated in the analysis and investigation of two species of Drosophila flies (D. melanogaster and D. simulans) to measure similarities and differences and their attraction to two different food substrates. Sleep and circadian measurements were also recorded during experimentation. The Drosophila Activity Monitor 5M (DAM5M) System and Sleep Circadian Analysis MATLAB Program (SCAMP) analysis were …

The Art Of Amphibian Conservation: Linking In-Situ And Ex-Situ Populations Of Endangered Species Through Genome Banking, Isabella Joann Burger

Theses and Dissertations

Limited breeding success in captive breeding programs has necessitated the development of assisted reproductive technologies (ART) to preserve and increase genetic variation and population numbers of both captive and wild amphibian groups. ART has been shown to be successful in numerous anuran species, and current studies focus on the application of ART in ex-situ populations. The focus of this project is to show that linking in-situ and ex-situ amphibian populations through sperm cryopreservation, genome banking, and in-vitro fertilization is possible, with the goal of increasing gene diversity throughout groups in order to produce self-sustaining, wild populations in the future. Specific …

Modulation Of Glucose Homeostasis By Nucleotide P2y2 Receptor And Biological Sex, Hailee Anne Marino

Graduate Theses/Dissertations

Recent insights into the pathological role of Nucleotide P2Y₂ receptor suggest P2Y₂R involvement in high fat diet-induced obesity and potentiates insulin resistance. However, these recent insights do not demonstrate how P2Y₂R modulates glucose homeostasis under physiological conditions. Further, it remains unknown how sex biological factors influence P2Y₂R receptor signaling in the regulation of glucose homeostasis. The research objective for the present study is to elucidate the novel roles of P2Y₂ in fasting blood glucose and glucose tolerance (basal insulin sensitivity) under resting conditions in males and females. We expected that under physiological …

Validation Of A Novel Trpm8 Knockout Mouse Model, Bilan Mohamed

Thinking Matters Symposium

Recent studies suggest that the use of thermoregulatory treatments that affect brown fat may help curb obesity. However it is unknown how these treatments may affect bone homeostasis. Our work has focused on the transient receptor potential melastatin (TRPM8) protein, which is responsible for detecting colder temperatures in sensory neurons. Previous work within the Motyl laboratory has found that Trpm8 plays a role in bone acquisition. Mice with a global deletion of the Trpm8 gene have reduced trabecular bone volume fraction due to reduced bone formation by osteoblasts. However, it is unclear whether sensory neuron or osteoblast-mediated expression of Trpm8 …

Morph- And Sex-Specific Differences In Corticosterone Of The Arizona Tiger Salamander (Ambystoma Mavortium Nebulosum), Megan Zerger

Scholars Week

Life history morph, sex, and body condition are traits that may influence stress within salamander populations because of differences in physiology and environmental conditions. Given widespread declines and the effects chronic stress can have on amphibian health, it is important to understand within-population drivers of stress and how population level variation may influence population viability. Thus, the objective of our study was to assess how corticosterone varies within the Arizona tiger salamander (Ambystoma mavortium nebulosum) population at the Mexican Cut Nature Preserve. We used a non-invasive skin swabbing method to collect baseline and elevated corticosterone from paedomorph (aquatic …

Vulnerable And Resilient Phenotypes In A Mouse Model Of Anorexia Nervosa, Jeff A. Beeler, Devry Mourra, Roseanna M. Zanca, Abigail Kalmbach, Celia Gellman, Benjamin Y. Klein, Rebecca Ravenelle, Peter Serrano, Holly Moore, Stephen Rayport, Susana Mingote, Nesha Burghard

Advanced Science Research Center

Background

Increased physical activity is a common feature of anorexia nervosa (AN). Although high activity levels are associated with greater risk of developing AN, particularly when combined with dieting, most individuals who diet and exercise maintain a healthy body weight. It is unclear why some individuals develop AN while most do not. A rodent model of resilience and vulnerability to AN would be valuable to research. Dopamine, which is believed to play a crucial role in AN, regulates both reward and activity and may modulate vulnerability.

Methods

Adolescent and young adult female C57BL/6N mice were tested in the activity-based anorexia …

Influence Of Testosterone On Male-Male Competition In The Red-Sided Garter Snake, Thamnophis Sirtalis Parietalis, Isabella M. G. Bukovich

Senior Honors Projects, 2020-current

Female mimicry in the red-sided garter snake, Thamnophis sirtalis parietalis, is described in several well-studied evolutionary advantages, but an aspect that remains elusive is how reproductive traits are linked to the 3-fold higher circulating androgens in males with this phenotype. In this study, we implanted male garter snakes (n=15 per group) with either a blank implant (SHAM; control) or a T implant, the latter simulating the female mimic phenotype. Following simulated low-temperature dormancy, males were scored for courtship intensity over three days of behavioral trials with females housed in the same facility. Males were sacrificed, and sperm were collected …

Respiratory Chain Activity And Protein Expression In Skeletal Muscles From High And Low Feed Efficient Swine Within The Same Genetic Line, Earsie L. Alcorn De Feliz

Graduate Theses and Dissertations

Current as well as past studies have provided evidence for a connection between macroeconomic/agricultural and cellular/biochemical basis of feed efficiency (FE; gain:feed) in farm animal production. Our investigation of respiratory chain activity, mitochondrial protein oxidation and protein expression was an expansion to the subcellular/molecular level. In this study we investigated the entire complex (Cx) of the electron transport chain (ETC) for differences in Cx activities in mitochondria isolated from skeletal muscle in high (HFE) and low (LFE) feed efficient swine within the same genetic line. Review of literature provided information for the connection between feed efficiency, mitochondrial function, and muscle …

Effect Of Ambient Temperature On Recovery Of Surgically Instrumented Sprague-Dawley Rats, Gianna Mangone

Senior Honors Projects, 2020-current

Based on current guidelines for housing rodents after surgical instrumentation, rodents may be housed at temperatures that hinder surgical recovery and cause thermal stress– room temperature (21°C) being one of them. Rodents are often housed at room temperature since this temperature is easy for humans to be caring for them. However, if recovering rodents are thermally stressed, experimental results will be confounded. To address this issue, Sprague-Dawley rats were surgically instrumented with radiotelemtry thermoprobes (Data Sciences, #TA-10F40) to monitor their core temperatures, then housed at one of five temperatures to assess the effect ambient temperature had on multiple aspects of …

Microglia-Neuron Interactions In A Mouse Model Of Low Grade Neuroepithelial Tumors, Veolette Hanna

Honors Scholar Theses

Microglia are the macrophages of the brain and spinal cord, playing an important role in the immune response to disease states of the nervous system. This study conducts an investigation on the activity of microglia in response to low grade neuroepithelial tumors. Using mouse models and microglial markers, a qualitative and quantitative analysis of microglia activation, migration, and invasion within the brain cortex during early stages of tumor development was conducted. It was found that the presence of a low grade neuroepithelial tumor in the cortex of one hemisphere of the brain causes significant microglia activation in comparison to the …

Ctrp3 And Alcoholic Liver Disease In Female Mice, Callie Root

Undergraduate Honors Theses

C1q TNF Related Protein 3 (CTRP3), is a cytokine that is primarily secreted from adipose tissue, which classifies it as an adipokine. Our previous research has shown that CTRP3 prevents alcoholic fatty liver disease (ALD) in male mice. However, even when accounting for confounding factors such as absolute and relative alcohol intake, females are more sensitive to the effects of consumption compared to male mice. Therefore, the goal of this project was to determine whether CTRP3 prevented ALD in female mice. Methods: Female wild type (WT) and female CTRP3 transgenic over expressing (Tg) mice were fed an ethanol containing liquid …

Genetic And Cellular Models For The Study Of Ptk2b And Adam10 In Adaptive Thermogenesis And Metabolism, Jared S. Farrar

Theses and Dissertations

In a cellular model, we demonstrate that the non-receptor protein tyrosine kinase 2 beta (PTK2B) plays a critical role in mouse cultured beige adipocyte differentiation. CRISPR/Cas9-mediated knock-out of Ptk2b results in non-differentiating white adipocytes and differentiated beige adipocytes with significantly reduced thermogenic gene and protein expression, enlarged lipid droplet size, and altered mitochondrial respiration. Together, these data in a cell culture system provide evidence for a role of PTK2B in the differentiation of mouse beige adipocytes.

In the process of developing a new mouse model utilizing the adipocyte selective Adipoq-Cre transgenic mouse, strong genetic linkage between a gene …

Focal Augmentation Of Somatostatin Interneuron Function And Subsequent Circuit Effects In Developmentally Malformed, Epileptogenic Cortex, Nicole Ekanem

Theses and Dissertations

Drug-resistant epilepsy (DRE) is a common clinical sequela of developmental cortical malformations such as polymicrogyria. Unfortunately, much remains unknown about the aberrant GABA-mediated circuit alterations that underlie DRE's onset and persistence in this context. To address this knowledge gap, we utilized the transcranial freeze lesion model in optogenetic mice lines (Somatostatin (SST)-Cre or Parvalbumin (PV)-Cre x floxed channelrhodopsin-2) to dissect features of the SST, PV, and pyramidal neuron microcircuit that are potentially associated with DRE. Investigations took place within developmental microgyria’s known pathological substrate, the adjoined and epileptogenic paramicrogyral region (PMR). As well, microcircuit relationships within the previously unexplored range …

Non-Invasive Method For Leptin Supplementation In Zebrafish (Danio Rerio), Regan Mcnamara

Williams Honors College, Honors Research Projects

I tested the hypothesis that recombinant leptin protein can be introduced to zebrafish in vivo through non-invasive soaking in a solution containing the protein. One way to study various molecules’ effects in vivo is through intraperitoneal or intracerebroventricular injections during the embryonic or larval stage, which is invasive, difficult to administer, and can have a high mortality rate. 48 hours post fertilization (hpf) zebrafish were soaked in a His-tagged recombinant leptin protein solution at 10 nM and 100 nM concentrations (produced by Genscript). After soaking, zebrafish larvae were washed extensively to remove all recombinant protein on their exterior before homogenization. …

Physiological Response Of Elasmobrachs During Propofol Immersion, Matthew Levendosky

College of Graduate Studies: Theses & Dissertations

Sensory experiments require anesthesia so the animal is immobilized, however fish anesthetics have shown to depress sensory responses. Newer anesthetics may offer similar anesthetic relief, but differ in means of action so sensory responses may be unaffected. Propofol has been used intravenously on small elasmobranchs but may provide prolonged effects if used as an immersion anesthetic. Objectives of this study were 1. Determine appropriate concentration of anesthetic to minimize induction and recovery for animals anesthetized at a surgical plane of anesthesia and 2. Measure physiological response of the pupil to light stimuli during anesthetic immersion. To address these objectives, I …

Tracing Nutrient Sources To Lipid Production In Insects Using Stable Isotope (Δ13c, Δ2h) Tracers: Implications For Nutritional Physiology Of Migratory Species., Libesha Anparasan, Keith A. Hobson

Western Research Forum

Using stable isotope measurements of insect tissues to determine origin and migratory patterns is well established. However, isotopically determining nutritional origins of lipids, the primary fuel of migration, has not been as thoroughly researched. We explored isotopic links between diet and stored lipids in laboratory raised True armyworm moths (Mythimna unipuncta) using δ¹³C and δ²H measurements. Pupae were randomly separated into four groups (n=20) and fed isotopically distinct nectar, each consisting of a combination of high δ¹³C (C4 sugar), or low δ¹³C (C3 sugar) carbohydrate, with high δ²H (deuterium spiked), …

The Effects Of Salinity On Zootechnical Performance Of Spotted Seatrout (Cynoscion Nebulosus) In Recirculating Aquaculture Systems, Eric Gigli

Master's Theses

The spotted seatrout (Cynoscion nebulosus) is a euryhaline finfish candidate for marine aquaculture in the northern Gulf of Mexico. Aquaculture of this species currently uses salinities of 25 psu or greater making production challenging in areas where high salinity water is not available. This study aimed to document the effects of salinity on zootechnical performance of spotted seatrout embryos, early larvae, and juveniles in recirculating aquaculture systems to assist with the development of protocols for low salinity culture.

Embryos obtained from a volitional spawn at 29 psu were incubated at three salinities (12.5, 18.75, or 25 psu) until …

Comparative Thermal Ecology Of Coastal And Inland Pacific Rattlesnakes (Crotalus Oreganus), Hayley Layne Crowell

Master's Theses

Global biodiversity is declining as a direct result of anthropogenic climate change. Ectothermic species have become focal organisms for studying the ecological effects of altered climates due to the clear relationship between environmental temperatures and ectotherms’ basic physiological functions. Historically, examinations of these effects have focused heavily on heliothermic lizards, and most others have tended to focus on single populations or sympatric species within a single community. Addressing the longterm energetic implications of environmental temperature variation will provide valuable insight into the cascading physiological effects that certain populations or species may experience as a result of altered climates.

In this …

Population Health Of Spotted Salamanders (Ambystoma Maculatum) In Created Vernal Pools: An Integrative Approach, Alice R. Millikin

Graduate Theses, Dissertations, and Problem Reports (ETD)

Habitat creation is an important tool for conservation to counteract habitat loss and degradation. Vernal pools are susceptible to destruction due to limited detection, protection, and regulation. These wetlands provide fishless breeding habitat for many amphibian species including spotted salamanders (Ambystoma maculatum) in eastern North America. Determining whether created vernal pool habitat is successful is often determined by demographic data of colonizing populations. I suggest that hormone levels, population genetics, and disease prevalence can improve our understanding of population health in created habitat. The goal of this dissertation was to assess the health of spotted salamander larvae in …

Cold Tolerance, Temperature Mediated Discontinuous Gas Exchange, And Emergence Of The Blue Orchard Mason Bee (Osmia Lignaria), Logan Kral

All Master's Theses

The relationship between low temperatures, emergence and supercooling point of Osmia lignaria were the subject of this study. One hundred sixty-eight bees were subjected to 5 pre-wintering temperature treatments (two constant temperature controls - 22^oC, 14^oC, one of which with and one without a 12h photoperiod, and three 12h:12h thermoperiod treatments – 14:10^oC, 14:5^oC, and 14:0^oC) and were then evaluated in terms of emergence time and post-emergence vigor. An additional 70 bees were tested for metabolic rate and discontinuous gas exchange in response to test temperature conditions. An additional sample …

Molecular Targets Of Psychedelics And Their Role In Behavioral Models Of Hallucinogenic Action, Hiba Z. Vohra

Theses and Dissertations

Psychedelics are a subset of hallucinogenic drugs that exert their characteristic effects through agonist activity at the serotonin receptor 2A (5-HT_2A). In this study, I aimed to characterize the modulatory role of the metabotropic glutamate subtype 2 receptor (mGluR2) in the 5-HT_2A-specific rodent model of hallucinogenic action, head-twitch response (HTR). Secondly, I aimed to explore if 5-HT_2A agonist-induced deficits in prepulse inhibition (PPI) of the startle response, an additional model of hallucinogenic action, could be produced in mice. Though 5-HT_2A agonist-induced PPI deficits, which represent interruptions in normal sensorimotor gating, have been described in …

Targeting Neuropeptides To Bone Fractures For Accelerated Healing, Nicholas A. Young, Jeffery J. Nielsen, Philip S. Low

The Summer Undergraduate Research Fellowship (SURF) Symposium

In patients over the age of 65 especially, bone fractures represent a significant disease burden. Non-invasive drug therapies are not available for bone fractures which represents a problem for this population. Vasoactive intestinal peptide (VIP) and Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP), two neuromodulator peptides in the glucagon superfamily, have demonstrated positive regulation of osteoblast proliferation and activity. Using acidic oligopeptides, we have developed ligands that target to and accumulate at fracture sites. These targeting ligands can be synthesized in sequence with bone anabolic peptides to minimize off target effects and increase potency at the fracture site to create safer and …

Pyruvate Kinase Isoform M2 Influences Autophagy And Related Processes In Hepatocellular Carcinoma Cells, Matthew Lin

University Scholar Projects

Hepatocellular carcinoma (HCC) is the most common form of liver cancer that affects ~14 million people in the world. Like all cancers, HCC is a disease that arises from unstinted cellular growth initiated by genetic alterations, metabolic changes, and dysregulation in key cellular pathways. Of interest is the relationship between metabolism and cell proliferation/degradation for therapeutic targeting. Pyruvate kinase M2 is a dimeric, glycolytically inactive isoform of the final enzyme involved in glycolysis, that is often upregulated in cancerous tissue. It is thought that the enzymatic function of PKM2 outside of glycolysis contributes to the biosynthesis of anabolic intermediates used …

Targeted Limb Heating Augments The Actions Of Igf1 In The Growth Plate And Increases Bone Elongation In Growing Mice, Holly Lyn Racine

Theses, Dissertations and Capstones

Bone elongation disorders can lead to painful musculoskeletal disabilities in adulthood. Existing treatment options to correct left-right asymmetry in limb length include invasive surgeries and/or drug regimens. These are often only partially effective. Previous studies in weanling mice have shown that a daily application of mild heat (40°C) to limbs on one side of the body could be used to noninvasively enhance bone elongation. However, the impact of heat-treatment on bone at the cellular level remains elusive. The epiphyseal growth plate, the band of cartilage located at each end of long bones, is the main site of longitudinal growth and …

Proteomic Analysis Of The Crustacean Molting Gland (Y-Organ) Over The Course Of The Molt Cycle, Talia B. Head

Master's Theses

Molting in crustaceans is a highly complex physiological process involving negative regulation by two paired endocrine glands, the X-organ/sinus gland complex (XO/SG) and the Y-organ (YO). The XO/SG complex is responsible for making molt-inhibiting hormone (MIH) which negatively regulates synthesis of the molting hormones, ecdysteroids, by the YO. Analysis of gene expression in the XOs and YOs has led to the development of a proposed molecular signaling pathway which regulates ecdysteroidogenesis and subsequent molting in crustaceans. In this study, changes in protein abundance in the YO were characterized over the course of a molt cycle (intermolt, early premolt, mid premolt, …