Animal Experimentation and Research Commons^™

A Native Function For Ran Translation And Cgg Repeats In Regulating Fragile X Protein Synthesis, Caitlin M. Rodriguez, Shannon E. Wright, Michael G. Kearse, Jill M. Haenfler, Brittany N. Flores, Yu Liu, Marius F. Ifrim, M. Rebecca Glineburg, Amy Krans, Paymaan Jafar-Nejad, Michael A. Sutton, Gary J. Bassell, Jack M. Parent, Frank Rigo, Sami J. Barmada, Peter K. Todd

Biology, Chemistry, and Environmental Sciences Faculty Articles and Research

Repeat-associated non-AUG-initiated translation of expanded CGG repeats (CGG RAN) from the FMR1 5′-leader produces toxic proteins that contribute to neurodegeneration in fragile X-associated tremor/ataxia syndrome. Here we describe how unexpanded CGG repeats and their translation play conserved roles in regulating fragile X protein (FMRP) synthesis. In neurons, CGG RAN acts as an inhibitory upstream open reading frame to suppress basal FMRP production. Activation of mGluR5 receptors enhances FMRP synthesis. This enhancement requires both the CGG repeat and CGG RAN initiation sites. Using non-cleaving antisense oligonucleotides (ASOs), we selectively blocked CGG RAN. This ASO blockade enhanced endogenous FMRP expression in human …

Activation Of The Sonic Hedgehog Effector Smoothened Counteracts L-Dopa Induced Dyskinesia By Restoring Cholinergic Interneuron Function, Lauren Malave

Dissertations, Theses, and Capstone Projects

Many types of neurons act as multimodal signaling centers. Yet, we have only limited insight into the regulation and functional consequences of neuronal co-transmission. For example, dopamine (DA) neurons, whose degeneration causes motor deficits characteristic of Parkinson’s Diseases (PD), communicate with all their targets by DA but only a selective subset of their targets using GABA, Glutamate, and the secreted cell signaling protein Sonic Hedgehog (Shh). It is unknown whether Levo-dopamine (L-Dopa) induced dyskinesia (LIDs), a severely debilitating side effect of DA supplementation in PD, might appear because DA neuron targets are exposed to high DA- but low Shh- signaling …

Demarcation Of Sepsis-Induced Peripheral And Central Acidosis With Ph (Low) Insertion Cycle Peptide, Kelly E. Henry, Aisling M. Chaney, Veronica L. Nagle, Haley C. Cropper, Saghar Mozaffari, Chip Slaybaugh, Keykavous Parang, Oleg A. Andreev, Yana K. Reshetnyak, Michelle L. James, Jason S. Lewis

Pharmacy Faculty Articles and Research

Acidosis is a key driver for many diseases, including cancer, sepsis, and stroke. The spatiotemporal dynamics of dysregulated pH across disease remain elusive, and current diagnostic strategies do not provide localization of pH alterations. We sought to explore if PET imaging using hydrophobic cyclic peptides that partition into the cellular membrane at low extracellular pH (denoted as pH [low] insertion cycles, or pHLIC) can permit accurate in vivo visualization of acidosis. Methods: Acid-sensitive cyclic peptide c[E₄W₅C] pHLIC was conjugated to bifunctional maleimide-NO2A and radiolabeled with ⁶⁴Cu (half-life, 12.7 h). C57BL/6J mice were administered lipopolysaccharide (15 …

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 …

Measuring The Effects Of Lobinaline-N-Bioxide (419) On Alcohol Consumption, Nicotine Locomotor Sensitization, And Conditioned Place Preference In Mice And Rats, Cocanut M. Suhail

Theses and Dissertations--Medical Sciences

Objective: Novel drug 419 was examined to see the effect it has in vivo mice and rats on alcohol consumption, nicotine locomotor sensitization, and conditioned place preference (CPP) models regarding behavioral tests on dopamine transporter activity.

Methods: Mice and rats were used to see how they react to the drug 419 and control vehicle, in each of the models. The animals were assessed to pre- and post- drug administration of novel drug 419. We examined each model to see the association between how drug 419 will help with treating drug abuse.

Results: We found that in alcohol consumption model the …

Frontal Brain Injury: Effects On Flexibility, Impulse Control, And Attention, Christopher Matthew O'Hearn

Graduate Theses, Dissertations, and Problem Reports (ETD)

Traumatic Brain Injury (TBI) is defined as an impact to the head, penetration of the skull, or rapid deceleration of the skull, resulting in an alteration of brain function or neurological deficit. Cognitive deficits are common following TBI and often go unresolved due to a lack of effective treatments. These deficits often perseverate into the chronic post injury phase, so the development of rehabilitative strategies is imperative. Behavioral flexibility, impulse control, and attention are a few cognitive processes that are commonly affected by TBI. The current research compares these processes between rats with and without a severe frontal brain injury …

Investigating The Morphology Of Iprgcs In Control Vs. Glaucomatous Retinas, Samantha Cook

Williams Honors College, Honors Research Projects

Glaucoma is a build-up of pressure in the eye that leads to the gradual loss of sight. The intention of this study was to investigate how glaucoma affects intrinsically photosensitive Retinal Ganglion Cells (ipRGCs). These cells contain melanopsin, a photopigment, that enables the cells to react to light. Four-month-old control and five-month-old glaucoma mice retinas were dissected, stained using primary and secondary antibodies, then traced using ImageJ’s Simple Neurite Tracer. There were no significant differences in soma size, total dendritic length, dendritic field size, and dendritic field diameter between control and glaucoma ipRGCs. However, further research is needed to provide …

Optimizing Animal Welfare In Commercial Laying Hens Through Novel Management Practices And Farm Manager Evaluation, John R. Brunnquell

Theses and Dissertations--Animal and Food Sciences

The commercial production of pasture and free-range eggs in the United States is expanding rapidly. According to the United States Department of Agriculture Agricultural Management Assistance, in May of 2019 there were 2.6M pasture hens and 4.5M free-range hens representing 1.9% of the nation’s laying flock. There is little industry and/or academic experience with this style of production in the United States. Programs such as the European Layer Training Initiative (ELTI), which emerged in 2019, have tried to fill this void but participation from the US was low. The principal sources for guidance of commercial free-range egg production in the …

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. …

Enhancing Chemosensitivity Of Lung Cancers By Manipulating The Tumor Microenvironment: Local Lung Delivery And Nanocarrier-Based Therapies, Hanming Zhang

Theses and Dissertations

Cancer is the second leading cause of death in United States causing an estimated six hundred thousand death in 2019. Among different types of cancer, lung cancer is the leading cause of cancer related death in both male and female. While targeted therapy and immunotherapy have shown promising clinical outcomes, currently there remain a significant proportion of patients who do not benefit from these strategies or who cannot tolerate them, making Pt based doublet chemotherapy still an indispensable component in almost all stages of treatment. However, partially due to the poor tumor distribution of small molecule drug, the development of …

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 …

Multivariate Analysis Of Open Field Exploration Identifies Latent Spatial And Social Behavioral Axes In Domestic Dogs, Budhaditya Chowdhury, Moira Van Staaden, Robert Huber

Advanced Science Research Center

Recent methodological advances in studying large scale animal movements have let researchers gather rich datasets from behaving animals. Often collected in small sample sizes due to logistical constraints, these datasets are however, ideal for multivariate explorations into behavioral complexity. In behavioral studies of domestic dogs, although automated data loggers have recently seen increasing use, a comprehensive framework to identify complex behavioral axes is lacking. Dog behavioral studies frequently rely on subjective ratings, despite demonstrable evidence that these are insufficient for identifying behavioral variables. Taking advantage of dogs’ innate running abilities and readily available GPS data loggers, we extracted latitude-longitude coordinates …

A Kinesin Adapter Directly Mediates Dendritic Mrna Localization During Neural Development In Mice, Hao Wu, Jing Zhou, Tianhui Zhu, Ivan Cohen, Jason Dictenberg

Publications and Research

Motor protein-based active transport is essential for mRNA localization and local translation in animal cells, yet how mRNA granules interact with motor proteins remains poorly understood. Using an unbiased yeast two–hybrid screen for interactions between murine RNA-binding proteins (RBPs) and motor proteins, here we identified protein interaction with APP tail-1 (PAT1) as a potential direct adapter between zipcode-binding protein 1 (ZBP1, a β-actin RBP) and the kinesin-I motor complex. The amino acid sequence of mouse PAT1 is similar to that of the kinesin light chain (KLC), and we found that PAT1 binds to KLC directly. Studying PAT1 in mouse …

Establishing An Ideal Inclusion Rate Of Fermented Soybean Meal And Sodium Butyrate On Growth Performance, Complete Blood Cell Count, And Nutrient Utilization In Nursery Pigs, Kristopher A. Bottoms

Graduate Theses and Dissertations

To evaluate increasing levels of sodium butyrate (SB) in nursery diets on growth performance (Experiment 1 & 2), complete blood cell count (Experiment 2), and the optimal level of fermented soybean meal for maximum performance in weanling pigs (Experiment 3), weaned pigs were blocked with initial body weight (BW) and allotted to dietary treatments. Treatments were: 1) Control (C) moderately complex corn-soybean-meal based supplemented with 0.05% benzoic acid (BA), but devoid of SB; C diet supplemented with 0.05%, 0.10%, or 0.15% SB (Experiment 1). Treatments in experiment 2 consisted of 1) a moderately complex corn-soybean-meal based diet devoid of SB …

Estimation Of Genetic Components Related To Infectious Bovine Keratoconjunctivitis Susceptibility In Angus And Angus Derived Cattle Produced In The Southern United States, Eric Oxford

Graduate Theses and Dissertations

The economic impact of infectious bovine keratoconjunctivitis (IBK) has been documented in many parts of the world. Many researchers have observed that prevention of this disease is very difficult given current methodologies. This is primarily due to the multifactorial nature of this disease. The objective of this dissertation was to determine the impact of IBK on calf performance and estimate genetic parameters, heritability and estimated breeding values for IBK susceptibility. Data were analyzed using PROC GLIMMIX of SAS; while genetic parameters were estimated using a linear animal model for both single- and two-traits through MTDFREML. Additional evaluations calculated heritability using …

Assessing Undergraduate Student And Faculty Views On Animal Research: What Do They Know, Whom Do They Trust, And How Much Do They Care?, Eric P. Sandgren, Robert Streiffer, Jennifer Dykema, Nadia Assad, Jackson Moberg

Attitudes Toward Animal Research Collection

Research using animals is controversial. To develop sound public outreach and policy about this issue, we need information about both the underlying science and people’s attitudes and knowledge. To identify attitudes toward this subject at the University of Wisconsin-Madison, we developed and administered a survey to undergraduate students and faculty. The survey asked respondents about the importance of, their confidence in their knowledge about, and who they trusted to provide information on animal research. Findings indicated attitudes varied by academic discipline, especially among faculty. Faculty in the biological sciences, particularly those who had participated in an animal research project, reported …

Phenotypically Plastic Responses To Predation Risk Are Temperature Dependent, Thomas M. Luhring, Janna M. Vavra, Clayton E. Cressler, John Delong

School of Biological Sciences: Faculty Publications

Predicting how organisms respond to climate change requires that we understand the temperature dependence of fitness in relevant ecological contexts (e.g., with or without predation risk). Predation risk often induces changes to life history traits that are themselves temperature dependent. We explore how perceived predation risk and temperature interact to determine fitness (indicated by the intrinsic rate of increase, r) through changes to its underlying components (net reproductive rate, generation time, and survival) in Daphnia magna. We exposed Daphnia to predation cues from dragonfly naiads early, late, or throughout their ontogeny. Predation risk increased r differentially across temperatures …

An Animal-Assisted Intervention Study In The Nursing Home: Lessons Learned, Lonneke G. J. A. Schuurmans, Inge Noback, Jos M. G. A. Schols, Marie-Jose Enders-Slegers

People and Animals: The International Journal of Research and Practice

AAI studies in the nursing home pose a specific set of challenges. In this article the practical and ethical issues encountered during a Dutch psychogeriatric nursing home AAI study are addressed with the aim of sharing our experiences for future researchers as well as AAI practitioners in general.

In our study we compared three groups of clients with dementia who participated in group sessions of either visiting dog teams, visiting FurReal Friend robot animals, or visiting students (control group) and monitored the effect on social interaction and neuropsychiatric symptoms through video analysis and questionnaires. We encountered the following four categories …

Effects Of Dabigatran In Mouse Models Of Aging And Cerebral Amyloid Angiopathy, Neethu Michael, Mher Mahoney Grigoryan, Kelley Kilday, Rachita K. Sumbria, Vitaly Vasilevko, Joanne Van Ryn, David H. Cribbs, Annlia Paganini-Hill, Mark J. Fisher

Pharmacy Faculty Articles and Research

Oral anticoagulants are a critical component of stroke prevention, but carry a risk of brain hemorrhage. These hemorrhagic complications tend to occur in elderly individuals, especially those with predisposing conditions such as cerebral amyloid angiopathy (CAA). Clinical evidence suggests that non-vitamin K antagonist oral anticoagulants are safer than traditional oral anticoagulants. We analyzed whether the anticoagulant dabigatran produces cerebral microhemorrhage (the pathological substrate of MRI-demonstrable cerebral microbleeds) or intracerebral hemorrhage in aged mice with and without hemorrhage-predisposing angiopathy. We studied aged (22 months old) Tg2576 (a model of CAA) and wild-type (WT) littermate mice. Mice received either dabigatran etexilate (DE) …

Adaptation Of The Systematic Review Framework To The Assessment Of Toxicological Test Methods: Challenges And Lessons Learned With The Zebrafish Embryotoxicity Test, Martin L. Stephens, Sevcan Gül Akgün-Ölmez, Sebastian Hoffman, Rob De Vries, Burkhard Flick, Thomas Hartung, Manoj Lalu, Alexandra Maertens, Hilda Witters, Robert Wright, Katya Tsaioun

Toxicology and Animal Models in Research Collection

Systematic review methodology is a means of addressing specific questions through structured, consistent, and transparent examinations of the relevant scientific evidence. This methodology has been used to advantage in clinical medicine, and is being adapted for use in other disciplines. Although some applications to toxicology have been explored, especially for hazard identification, the present preparatory study is, to our knowledge, the first attempt to adapt it to the assessment of toxicological test methods. As our test case, we chose the zebrafish embryotoxicity test (ZET) for developmental toxicity and its mammalian counterpart, the standard mammalian prenatal development toxicity study, focusing the …

Animal Research, Accountability, Openness And Public Engagement: Report From An International Expert Forum, Elisabeth H. Ormandy, Daniel M. Weary, Katarina Cvek, Mark Fisher, Kathrin Herrmann, Pru Hobson-West, Michael Mcdonald, William Milsom, Margaret Rose, Andrew Rowan, Joanne Zurlo, Marina A.G. Von Keyserlingk

Oversight of Animal Experimentation Collection

In November 2013, a group of international experts in animal research policy (n = 11) gathered in Vancouver, Canada, to discuss openness and accountability in animal research. The primary objective was to bring together participants from various jurisdictions (United States, Sweden, Australia, New Zealand, Germany, Canada and the United Kingdom) to share practices regarding the governance of animals used in research, testing and education, with emphasis on the governance process followed, the methods of community engagement, and the balance of openness versus confidentiality. During the forum, participants came to a broad consensus on the need for: (a) evidence-based metrics to …

Characterization Of The Swine Gut Microbiome Dynamics, Culturomics And Probiotics, Xiaofan Wang

Graduate Theses and Dissertations

The gastrointestinal tract is a critical place where the immune system concentrates many of its protective functions. It exerts both physical and biochemical defenses against invading foodborne pathogens and other hazards. In pigs, the gut microbiota community assembled postnatal plays a very important role in the development of multiple physiologic aspects, such as development of the gastrointestinal tract, nutrient exchange, immune defense against foreign pathogens and growth performance. Microbiome-host communication changes with age. Despite recent advances in understanding the swine gut microbiome at different growth stages, a comprehensive longitudinal study of the lifelong dynamics of the swine gut microbiome is …

Short Amylin Receptor Antagonist Peptides Improve Memory Deficits In Alzheimer’S Disease Mouse Model, Rania Soudy, Ryoichi Kimura, Aarti Patel, Wen Fu, Kamaljit Kaur, David Westaway, Jing Yang, Jack Jhamandas

Pharmacy Faculty Articles and Research

Recent evidence supports involvement of amylin and the amylin receptor in the pathogenesis of Alzheimer’s disease (AD). We have previously shown that amylin receptor antagonist, AC253, improves spatial memory in AD mouse models. Herein, we generated and screened a peptide library and identified two short sequence amylin peptides (12–14 aa) that are proteolytically stable, brain penetrant when administered intraperitoneally, neuroprotective against Aβ toxicity and restore diminished levels of hippocampal long term potentiation in AD mice. Systemic administration of the peptides for five weeks in aged 5XFAD mice improved spatial memory, reduced amyloid plaque burden, and neuroinflammation. The common residue SQELHRLQTY …

Contribution Of Animal Models To Contemporary Understanding Of Attention Deficit Hyperactivity Disorder, Constança Carvalho, Mariana Vieira Crespo, Luísa Ferreira Bastos, Andrew Knight, Luís Vincente

Biomedicine and Animal Models in Research Collection

Attention Deficit Hyperactivity Disorder (ADHD) is a poorly understood neurodevelopmental disorder of multifactorial origin. Animal-based research has been used to investigate ADHD etiology, pathogenesis and treatment, but the efficacy of this research for patients has not yet been systematically evaluated. Such evaluation is important given the resource consumption and ethical concerns incurred by animal use.

We used the citation tracking facility within Web of Science to locate citations of original research papers on animal models related to ADHD published prior to 2010 identified in PubMed by relevant search terms. Human medical papers citing those animal studies were carefully analyzed by …

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), …

Automated Monitoring Of Behaviour In Zebrafish After Invasive Procedures, Anthony G. Deakin, Jonathan Buckley, Hamzah S. Alzu'bi, Andrew R. Cossins, Joseph W. Spencer, Waleed Al'nuaimy, Iain S. Young, Jack S. Thomson, Lynne U. Sneddon

Validation of Animal Experimentation Collection

Fish are used in a variety of experimental contexts often in high numbers. To maintain their welfare and ensure valid results during invasive procedures it is vital that we can detect subtle changes in behaviour that may allow us to intervene to provide pain-relief. Therefore, an automated method, the Fish Behaviour Index (FBI), was devised and used for testing the impact of laboratory procedures and efficacy of analgesic drugs in the model species, the zebrafish. Cameras with tracking software were used to visually track and quantify female zebrafish behaviour in real time after a number of laboratory procedures including fin …

Cyclin C Regulated Oxidative Stress Responsive Transcriptome In Mus Musculus Embryonic Fibroblasts, David C Stieg, Kai-Ti Chang, Katrina F Cooper, Randy Strich

Rowan-Virtua School of Osteopathic Medicine Departmental Research

The transcriptional changes that occur in response to oxidative stress help direct the decision to maintain cell viability or enter a cell death pathway. Cyclin C-Cdk8 is a conserved kinase that associates with the RNA polymerase II Mediator complex that stimulates or represses transcription depending on the locus. In response to oxidative stress, cyclin C, but not Cdk8, displays partial translocation into the cytoplasm. These findings open the possibility that cyclin C relocalization is a regulatory mechanism governing oxidative stress-induced transcriptional changes. In the present study, the cyclin C-dependent transcriptome was determined and compared to transcriptional changes occurring in oxidatively …

Development Of A Pediatric Model Of Nafld In Neonatal Iberian Pigs, Gabriella Veronica Hernandez, Victoria Alice Smith, Morgan Coffin, Daniel Columbus, Matthew Burd, Kimberly Sprayberry, Mark Edwards, Daniel Peterson, Darin Bennett, Robert Fanter, Christopher Kitts, Michael La Frano, Margaret Rice, Douglas Burrin, Magdalena Maj, Rodrigo Manjarin

Master's Theses

The prevalence of non-alcoholic fatty liver disease (NAFLD) in children has increased over the past decades, creating a need for animal models that recapitulate the features of the pediatric disease. Iberian pigs have a leptin-resistant phenotype characterized by hyperleptinemia, hyperphagia, and extreme adipogenesis. We hypothesized that neonatal Iberian pigs fed a high fat high-fructose (HFF) diet will develop a pattern of liver injury resembling pediatric NAFLD. In addition, we sought to determine if a mixture of probiotics would prevent the disease. Animals were fed 1 of 4 diets containing (g/kg body weight × d) 0 g fructose, 11 g fat …

A Quiet Revolution Replacing The Use Of Animals In Research, Andrew N. Rowan

WellBeing News

In May, 2019, the Sanger Institute, a major producer of genetically modified mice announced it was closing down its GMO mouse unit. They came to this decision “following a rigorous review” of its scientific strategy and after consulting with the Wellcome Trust, one of the major funders of biomedical research in the world today and a supporter of programs at the Sanger Institute. This decision presages a new phase in animal research when animal use will again begin to decline as other technologies take the place of animal studies in major laboratories.