Life Sciences Commons^™

Pdz Protein Regulation Of Β-Arrestin Recruitment And Gpcr Trafficking, Sarah Gupta

Electronic Thesis and Dissertation Repository

β-arrestins are versatile adaptor proteins that play a vital role in regulation of G protein coupled receptor (GPCR) trafficking and signalling properties. PDZ proteins have previously been shown to modulate β-arrestin2 recruitment and receptor internalization for many GPCRs including Corticotropin-Releasing Factor Receptor 1 (CRFR1), a receptor whose antagonists have been shown to demonstrate both anxiolytic- and antidepressant-like effects. Further characterization of the interplay between β-arrestins and PDZ proteins may aid in determining a potential mechanism for PDZ protein regulation of GPCR trafficking. Our findings suggest that PDZ proteins PSD-95, MAGI1, and PDZK1 complex with β-arrestin2 by interacting via the PDZ …

New Insights Into Signal Detection Of The Effects Of Exposures During Pregnancy, Fatma Etwel

Electronic Thesis and Dissertation Repository

There is inadequate information on the fetal safety of drugs during pregnancy for the majority of marketed drugs. It is challenging to examine the safety and efficacy of drugs during pregnancy due to the ethical issues of exposing unborn babies to these chemicals. It often takes many years before associations between a drug and its safety, efficacy, and toxicity in pregnancy can be established. This thesis will examine strategies in signal detection of the effects of drug exposures during pregnancy.

Meta-analyses have become useful in the area of clinical teratology. Observational studies provide the main source of information in these …

Evidence Of A Role For G Protein Signaling Modulator 3 In Cell Division, Drew C. Wallace

Electronic Thesis and Dissertation Repository

Components of G protein-mediated signaling are associated with positioning and orienting the mitotic spindle in the process of cell division. However, a functional role for G protein signaling modulator 3 (GPSM3) in cell division has yet to be defined. The purpose of this study was to investigate a potential role for GPSM3 in cell division. Vascular smooth muscle cells (VSMCs) from Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats, that are known to express GPSM3, were used as a model system. Here I report that GPSM3 mRNA and protein levels varied during different stages of the cell cycle in SHR VSMCs. …

The Association Between Oxidative Stress, Cellular Differentiation And Galectins In Human Promyelocytic Leukemia Cells (Hl-60), James R. Vinnai

Electronic Thesis and Dissertation Repository

Galectins are a group of β-galactoside-binding proteins involved in different cellular processes including stress responses and differentiation. The role and expression of galectins under oxidative stress and during neutrophilic differentiation was examined in HL-60 cells. Galectin gene (LGALS), and galectin protein expression were determined using RT-qPCR and immunoblotting, respectively. Neutrophilic differentiation was measured via a spectrofluorometric assay. DNA methylation and JNK signaling were investigated as galectin regulatory mechanisms. Menadione-induced oxidative stress, DMSO-induced differentiation, DNA hypomethylation and JNK signaling all promoted similar galectin expression profiles. Antioxidant N-acetylcysteine attenuated the menadione-induced galectin expression but only partially attenuated DMSO-induced galectin expression. …

Producing A Subunit Vaccine For Porcine Epidemic Diarrhea Virus, Zayn Khamis

Electronic Thesis and Dissertation Repository

Porcine epidemic diarrhea virus (PEDv) causes disease and mortality to piglets worldwide. Most vaccines used to combat the disease have been ineffective live attenuated virus vaccines. The goal of this project was to produce a plant-made subunit vaccine based off the membrane protein of the virus. This is the first time this protein has been produced in plants. An elastin-like polypeptide fusion membrane protein accumulated up to 0.8 mg/g of fresh leaf weight when transiently expressed in Nicotiana benthamiana. Virus-like particles were also produced for the first time for PEDv, and were able to form with just the membrane …

Comprehensive Molecular Characterization Of Human Nodal, Scott D. Findlay

Electronic Thesis and Dissertation Repository

Nodal and related ligands are highly conserved members of the TGF-beta superfamily with well-established and essential roles in the early embryonic development of vertebrates, and in cell fate decisions in human embryonic stem (hES) cells. Aberrant NODAL signaling also generally promotes pro-tumourigenic phenotypes and the progression of a wide array of human cancers. Despite being pursued as a potential therapeutic target, many aspects of NODAL’s molecular biology remain poorly understood. This thesis provides a comprehensive characterization of gene expression from the human NODAL locus at multiple levels. First, an intronic NODAL SNP known as rs2231947 was found to be functional …

Insights Into The Role Of Connexin26 And Connexin30 In Skin Health, Syndromic Disease, And Cutaneous Wound Healing, Eric R. Press

Electronic Thesis and Dissertation Repository

Connexin26 (Cx26) and Cx30 facilitate gap junctional intercellular communication (GJIC) in the epidermis and are linked to several syndromic skin diseases. We investigated 5 disease- linked Cx26 mutants and demonstrated that the severity and extent of disease can be predicted from the gain- or loss-of function properties of each mutant, as well as the ability to induce cell death. We also used transgenic mice expressing S17F Cx26 (linked to keratitis-ichthyosis- deafness syndrome) or A88V Cx30 (linked to Clouston Syndrome) to investigate the pathophysiology these skin diseases. We demonstrated that S17F Cx26, but not A88V Cx30, promotes palmoplantar keratoderma by disrupting …

Riboregulation Of Bacterial Transposons, Michael J. Ellis

Electronic Thesis and Dissertation Repository

Bacterial transposons typically exist in a mutually beneficial relationship with the host cell. Limited transposition can benefit the host while also ensuring the survival of the element. An important component of this relationship is that transposition must be tightly regulated. In this thesis I explore ways that the host and transposon each control transposition in E. coli and provide evidence that a transposon can also control host gene expression in S. enterica Typhimurium. Post-transcriptional regulation with small non-coding RNAs (sRNA) has emerged as a key way that bacteria respond to stress and regulate many cellular processes. The RNA-binding protein Hfq …

The Impacts Of Climate Change On Communities Of Fungi In Boreal Peatlands, Asma Asemaninejad Hassankiadeh

Electronic Thesis and Dissertation Repository

Peatlands have an important role in global climate change through sequestration of atmospheric CO₂. Climate change is already affecting these ecosystems, including both above- and below- ground communities and their functions. Fungi play a central role in these communities. As a result, there is concern that altered fungal community function may turn peatlands from carbon sinks to carbon sources, greatly exacerbating the impacts of climate change. In order to gain a better insight into effects of climate change on the structure and function of these carbon sequestrating ecosystems, this thesis focuses on diversity and structure of fungal communities …

Effects Of Gut-Associated Yeasts On Drosophila Melanogaster Performance, Yanira Jiménez Padilla

Electronic Thesis and Dissertation Repository

I used Drosophila melanogaster as a model to study the role of the gut microbiota, specifically yeasts, in animal physiology. I used Saccharomyces cerevisiae, the yeast commonly included in Drosophila diet, and Lachancea kluyveri, isolated from some Drosophila in the wild, and generated axenic (germ-free) and gnotobiotic (yeast-fed) flies. I found that L. kluyveri persists in the crop, as ascospores and vegetative cells, longer than S. cerevisiae. Some L. kluyveri vegetative cells survive passage through the gut. Egg to adult development time is reduced by 14% in vials containing live L. kluyveri or S. cerevisiae, whereas …

Freeze Tolerance Of Cyphoderris Monstrosa (Orthoptera: Prophalangopsidae), Jantina Toxopeus, Jacqueline Lebenzon, Alexander H. Mckinnon, Brent J. Sinclair

Biology Publications

The great grig, Cyphoderris monstrosa Uhler (Orthoptera: Prophalangopsidae), is a large (20-30 mm, >1 g), nocturnal ensiferan that in habits montane coniferous forests in northwestern North America. C. monstrosa overwinters as a late-instar nymph, but its cold tolerance strategy has not previously been reported. We collected nymphs from near Kamloops, British Columbia, in late spring to determine their cold tolerance strategy. C. monstrosa nymphs were active at low temperatures until they froze at -4.6 ± 0.3 °C. The nymphs survived internal ice formation (i.e. are freeze tolerant), had a lethal temperature between -9 and -12 °C, and could survive for …

Characterization Of Ranbpm Subcellular Localization And Function In Hdac6 Regulation, Louisa M. Salemi

Electronic Thesis and Dissertation Repository

RanBPM has been shown to interact with numerous proteins implicating it in a variety of cellular processes including apoptosis, transcription regulation, cell migration, adhesion and morphology and has been shown to have tumour suppressive functions. RanPBM has been hypothesized to be a scaffolding protein and is part of a large protein complex termed the CTLH complex. Although the homologous complex in yeast has been demonstrated to have E3 ubiquitin ligase activity, whether the evolutionarily conserved human complex retains this activity remains to be determined. In this work we aim to characterize the regions of RanBPM that regulate its subcellular localization …

Epithelial-To-Mesenchymal Transition Leads To Disease-Stage Differences In Circulating Tumor Cell Detection And Metastasis In Pre-Clinical Models Of Prostate Cancer, Lori E. Lowes, David Goodale, Ying Xia, Carl Postenka, Matthew M. Piaseczny, Freeman Paczkowski, Alison L. Allan

Anatomy and Cell Biology Publications

Metastasis is the cause of most prostate cancer (PCa) deaths and has been associated with circulating tumor cells (CTCs). The presence of >= 5 CTCs/7.5mL of blood is a poor prognosis indicator in metastatic PCa when assessed by the CellSearch (R) system, the "gold standard" clinical platform. However, similar to 35% of metastatic PCa patients assessed by CellSearch (R) have undetectable CTCs. We hypothesize that this is due to epithelial-to-mesenchymal transition (EMT) and subsequent loss of necessary CTC detection markers, with important implications for PCa metastasis. Two pre-clinical assays were developed to assess human CTCs in xenograft models; one comparable …

Impact Of Extremely Low-Frequency Magnetic Fields On Human Postural Control, Alicia N. Allen

Electronic Thesis and Dissertation Repository

The general public and workers can be exposed to high-levels of power-line frequency magnetic fields (MFs - up to 10 mT). Although such time-varying MFs have the potential to modulate human postural control, no existing studies have explored MF exposure levels that possibly trigger acute sway responses. This work evaluates time-varying MF exposure (up to 100 mT) in the extremely low frequency range (ELF – up to 300 Hz) and its effects on human postural control. Twenty-two healthy participants were each exposed to randomized, 5-second MF and electric stimulations (0, 50 and 100 mT and 1.5 mA respectively) given at …

Species Pluralism: Conceptual, Ontological, And Practical Dimensions, Justin Bzovy

Electronic Thesis and Dissertation Repository

Species are central to biology, but there is currently no agreement on what the adequate species concept should be, and many have adopted a pluralist stance: different species concepts will be required for different purposes. This thesis is a multidimensional analysis of species pluralism. First I explicate how pluralism differs monism and relativism. I then consider the history of species pluralism. I argue that we must re-frame the species problem, and that re-evaluating Aristotle's role in the histories of systematics can shed light on pluralism. Next I consider different forms of pluralism: evolutionary and extra-evolutionary species pluralism, which differ in …

Regulation Of E2f1 In Keratinocytes During Uv-Damage And Differentiation, Randeep K. Singh

Electronic Thesis and Dissertation Repository

The E2F1 transcription factor regulates the expression of key genes involved in cell proliferation and differentiation to maintain skin homeostasis. The expression of E2F1 is tightly regulated during cell cycle progression and when cells are committed to differentiate, as well as in response to DNA damage. In keratinocytes, E2F1 protein and transcript levels increase following UV-induced DNA damage, whereas, in response to Ca2+-induced differentiation, both E2F1 protein and transcript levels decrease. In this thesis, I examined in detail the mechanism that modulates E2F1 stability following DNA damage and during keratinocyte differentiation. I show that E2F1 associates with hHR23 and together …

The Use Of P3b As An Indicator Of Neurophysiologic Change From Subconcussive Impacts In Football Players, Jeffrey S. Brooks

Electronic Thesis and Dissertation Repository

There is a growing appreciation in research that subconcussive impacts may affect cognitive functioning. Canadian University football players (n=45) were separated into three groups based on their position/skill (small skilled, big skilled and big unskilled). An impact measuring device (GForceTracker) was used to record the number of impacts that each player experienced in a season. Player groups were separated into two levels of impact exposure: low and high. Players completed baseline, midseason, postseason, and follow-up neurophysiological tests (four months later) to measure P3b amplitude in response to a visual oddball paradigm, and high versus low impact subgroups for each player …

A Rat Model Of Cognitive Bias And The Effect Of Acute Corticosterone, Kai Wang

Electronic Thesis and Dissertation Repository

In humans, affective states can influence cognitive processes, resulting in a phenomenon referred to as “cognitive bias”. Rodents exhibit similar biases during the interpretation of ambiguous cues. It has been shown that cognitive bias shifts towards the negative valence (pessimism) when animals are under chronic stress manipulations. However, the effects of acute stress on cognitive bias have not been well established in an animal model. Here, a non-operant appetitive task using fluid rewards and distinct visual/tactile cues was developed to examine cognitive bias in male rats. Corticosterone was used to mimic stress levels similar to acute restraint stress. It was …

Can We Predict Ectotherm Responses To Climate Change Using Thermal Performance Curves And Body Temperatures?, Brent J Sinclair, Katie E Marshall, Mary A Sewell, Danielle L Levesque, Christopher S Willett, Stine Slotsbo, Yunwei Dong, Christopher D G Harley, David J Marshall, Brian S Helmuth, Raymond B Huey

Biology Publications

Thermal performance curves (TPCs), which quantify how an ectotherm's body temperature (Tb ) affects its performance or fitness, are often used in an attempt to predict organismal responses to climate change. Here, we examine the key - but often biologically unreasonable - assumptions underlying this approach; for example, that physiology and thermal regimes are invariant over ontogeny, space and time, and also that TPCs are independent of previously experienced Tb. We show how a critical consideration of these assumptions can lead to biologically useful hypotheses and experimental designs. For example, rather than assuming that TPCs are fixed during ontogeny, one …

Corticotropin Releasing Factor Receptor Type 1 Signaling In Epilepsy And Traumatic Brain Injury, V V Chakravarthi Narla

Electronic Thesis and Dissertation Repository

Stress increases the frequency by which epileptic seizures occur. Corticotropin-releasing factor (CRF) coordinates neuroendocrine, autonomic and behavioral response to stress. This thesis sought to study the cellular and molecular mechanisms by which CRF regulates the activity of neural circuits in the piriform cortex (PC) in normal and epileptic states. The PC is richly innervated by CRF and 5-HT containing axons arising from the central amygdala and raphe nucleus. CRFR₁ and 5-HT_2A/CRs have been shown to interact in a manner where CRFR activation subsequently potentiates the activity of 5-HT_2A/CRs. The first purpose of this thesis was …

Molecular Characterization Of Viruses Infecting Greenhouse Vegetables In Ontario, Bin Chen

Electronic Thesis and Dissertation Repository

Greenhouse vegetable production is a very important and successful agricultural sector in Ontario. However, newly emerging viral or virus-like diseases have become a major limiting factor in greenhouse vegetable production. To determine the identity of the viral pathogens in the three major greenhouse vegetables in Ontario, we conducted in-depth sequencing of small RNAs isolated from virally infected plants used next-generation sequencing technology. Subsequent bioinformatics analyses revealed six viral pathogens including Bell pepper endornavirus (BPEV), Parietaria mottle virus (PMoV), Cucumber green mottle mosaic virus (CGMMV), Tobacco streak virus (TSV), Pepper mild mottle virus (PMMoV), and Pepino mosaic virus (PepMV). The full-length …

The Role Of Cholinergic Neurotransmission In Sensory Filtering And Sensorimotor Gating, Erin Azzopardi

Electronic Thesis and Dissertation Repository

At every moment, our brain is bombarded with sensory information. How we filter and process sensory information is critical for daily functioning and cognition. Examples of sensory filtering include habituation (a progressive decrease in responding) and prepulse inhibition (PPI, gating of responding). Our aim is to understand the differential role acetylcholine (ACh) plays in these processes.

To study this we used both reflexive (acoustic startle response: ASR) and non-reflexive (locomotor) behaviours. PPI is hypothesized to occur via inhibitory cholinergic projections from the Pedunculopontine Tegmental Nucleus (PPT) to the startle pathway. The role of ACh in habituation of reflexive and non-reflexive …

The Effects Of Ice Hockey Goaltender Leg Pads On Safety And Performance, Ryan J. Frayne

Electronic Thesis and Dissertation Repository

Ice hockey goaltenders have the highest percentage of cam-type femoroacetabular impingement (FAI). The exact cause of these injuries in goaltenders remains unknown; however, it has been suggested that common goaltender movements and a goaltender’s underlying hip pathology may be contributing factors. The butterfly save technique, commonly used by goaltenders, has been linked to FAI. Simply stopping these movements would likely be detrimental to goaltender performance. Therefore, changing other aspects of goaltending, such as altering the goaltender equipment, should be considered. The overall objective of this thesis was to understand how ice hockey goaltender leg pads influence both the safety and …

Winter Warming Effects On Overwinter Survival, Energy Use, And Spring Emergence Of Cerotoma Trifurcata (Coleoptera: Chrysomelidae), Emily A. Berzitis, Heather A. Hager, Brent J Sinclair, Rebecca H. Hallett, Jonathan A. Newman

Biology Publications

1. Bean leaf beetle Cerotoma trifurcata (Förster) (Coleoptera: Chrysomelidae) is a pest of soybean in the U.S.A. and is becoming a concern in Canada. The projected increase in winter temperatures under climate change could affect overwinter survival, timing of spring emergence and, ultimately, the severity of this pest.
2. We assessed the potential effects of warmer winters in field experiments performed in three consecutive years. Three warming levels were applied: (i) heated approximately 4 °C above ambient; (ii) unheated with snow cover left intact; and (iii) unheated with snow cover removed. Survival and date of emergence were assessed in all years, and …

Regulation Of Amyloid Oligomer Binding To Neurons And Neurotoxicity By The Prion Protein-Mglur5 Complex, Flavio H. Beraldo, Valeriy G. Ostapchenko, Fabiana A. Caetano, Andre L. S. Guimaraes, Giulia D. S. Ferretti, Nathalie Daude, Lisa Bertram, Katiane O. P. C. Nogueira, Jerson L. Silva, David Westaway, Neil R. Cashman, Vilma R. Martins, Vania F. Prado, Marco A. M. Prado

Anatomy and Cell Biology Publications

The prion protein (PrPC) has been suggested to operate as a scaffold/receptor protein in neurons, participating in both physiological and pathological associated events. PrPC, laminin, and metabotropic glutamate receptor 5 (mGluR5) form a protein complex on the plasma membrane that can trigger signaling pathways involved in neuronal differentiation. PrPC and mGluR5 are co-receptors also for -amyloid oligomers (AOs) and have been shown to modulate toxicity and neuronal death in Alzheimer's disease. In the present work, we addressed the potential crosstalk between these two signaling pathways, laminin-PrPC-mGluR5 or AO-PrPC-mGluR5, as well as their interplay. Herein, we demonstrated that an existing complex …

Cell Signaling Crosstalk And Differentiation Of F9 Cells Into Extraembryonic Endoderm, Gurjoth Singh Deol

Electronic Thesis and Dissertation Repository

Mouse F9 cells differentiate into primitive endoderm (PrE) when treated with retinoic acid (RA). During PrE differentiation the canonical Wnt signaling pathway is known to play an integral role in the process. In addition to Wnt signaling, there have been implications that the Hedgehog (Hh) pathway may also be involved. Previous results show that the Indian Hedgehog (Ihh) gene is upregulated during RA-induced differentiation; however further details of Hh signaling during PrE differentiation have yet to be discovered. A Gli-luciferase construct indicated that Hh signaling increases during RA-induced differentiation, implicating that the pathway is involved in PrE formation. Inhibiting …

Vacht Overexpression Increases Acetylcholine At The Synaptic Cleft And Accelerates Aging Of Neuromuscular Junctions, Satoshi Sugita, Leland L. Fleming, Caleb Wood, Sydney K. Vaughan, Matheus P. S. M. Gomes, Wallace Camargo, Ligia A. Naves, Vania F. Prado, Marco A. M. Prado, Cristina Guatimosim, Gregorio Valdez

Anatomy and Cell Biology Publications

Background: Cholinergic dysfunction occurs during aging and in a variety of diseases, including amyotrophic lateral sclerosis (ALS). However, it remains unknown whether changes in cholinergic transmission contributes to age-and disease-related degeneration of the motor system. Here we investigated the effect of moderately increasing levels of synaptic acetylcholine (ACh) on the neuromuscular junction (NMJ), muscle fibers, and motor neurons during development and aging and in a mouse model for amyotrophic lateral sclerosis (ALS). Methods: Chat-ChR2-EYFP (VAChTHyp) mice containing multiple copies of the vesicular acetylcholine transporter (VAChT), mutant superoxide dismutase 1 (SOD1G93A), and Chat-IRES-Cre and tdTomato transgenic mice were used in this …

Editorial Overview: Global Change Biology: Linking Pattern And Process To Prediction And Policy., Vladimir Koštál, Brent J Sinclair

Biology Publications

No abstract provided.

Can We Predict The Effects Of Multiple Stressors On Insects In A Changing Climate?, Sirpa Kaunisto, Laura V Ferguson, Brent J Sinclair

Biology Publications

The responses of insects to climate change will depend on their responses to abiotic and biotic stressors in combination. We surveyed the literature, and although synergistic stressor interactions appear common among insects, the thin taxonomic spread of existing data means that more multi-stressor studies and new approaches are needed. We need to move beyond descriptions of the effects of multiple stressors to a mechanistic, predictive understanding. Further, we must identify which stressor interactions, and species' responses to them, are sufficiently generalizable (i.e. most or all species respond similarly to the same stressor combination), and thus predictable (for new combinations of …

Preterm Neonatal Lateral Ventricle Volume From Three-Dimensional Ultrasound Is Not Strongly Correlated To Two-Dimensional Ultrasound Measurements, Jessica Kishimoto, Sandrine De Ribaupierre, Fateme Salehi, Walter Romano, David S. C. Lee, Aaron Fenster

Anatomy and Cell Biology Publications

The aim of this study is to compare longitudinal two-dimensional (2-D) and three-dimensional (3-D) ultrasound (US) estimates of ventricle size in preterm neonates with posthemorrhagic ventricular dilatation (PHVD) using quantitative measurements of the lateral ventricles. Cranial 2-D US and 3-D US images were acquired from neonatal patients with diagnosed PHVD within 10 min of each other one to two times per week and analyzed offline. Ventricle index, anterior horn width, third ventricle width, and thalamo-occipital distance were measured on the 2-D images and ventricle volume (VV) was measured from 3-D US images. Changes in the measurements between successive image sets …