Life Sciences Commons^™

The Biobehavioral Effects Of Embryonic Exposure To Neural Inflammation And Oxidative Stress In Zebrafish, Dalton J. Anderson

Honors College Theses

The purpose of this research is to improve understanding of the neurodevelopmental effects of embryonic exposure to elevated inflammation and oxidative stress induced by the antipyretic drug acetaminophen (APAP). Our study was the first to examine the interactive effects of APAP and inflammation in zebrafish embryos and how the treatments affect brain development and larval behavior. Experimental groups of zebrafish larvae were exposed to lipopolysaccharide (LPS) to induce inflammation, APAP, or LPS + APAP and larval behavior was analyzed using Ethovision automated behavioral tracking software. We also measured changes in whole-brain Glycogen Synthase Kinase 3 Beta (GSK3B) and GSK3B phosphorylation, …

Zebrafish Electroretinogram Responses, Brooke Campbell

Williams Honors College, Honors Research Projects

The goal of this project is to be able to streamline a protocol for conducting ERGs on zebrafish from mice ERG protocol already present in Dr. Renna’s lab. The objective is to create a protocol specifically for zebrafish and to eliminate any issues that occur when transiting from a mice ERG protocol to a zebrafish ERG protocol and to obtain a light response from zebrafish retinae in differing light intensities. Dr. Renna’s lab has designed an Ex Vivo ERG system with less electrical interference creating defined electrical responses from mouse retina. The setup allows for continual perfusion of the retinal …

Roles Of Ppp1r15a (Gadd34) And Ppp1r15b (Crep) In Er Stress-Induced Apoptosis In Zebrafish Caudal Fin Epidermal Cells, Ayano Ohata

University of the Pacific Theses and Dissertations

ER stress occurs in response to the accumulation of unfolded or misfolded proteins in the ER lumen, subsequently activating three signal transduction pathways collectively called the unfolded protein response (UPR). Although the goal of the UPR is to restore ER homeostasis, it can result in apoptosis when ER stress is too severe or prolonged. CHOP, which is induced by all three branches of the UPR, is central to inducing ER stress-induced apoptosis by regulating the expression of numerous pro-apoptotic genes. One of the downstream targets of CHOP is GADD34, which contributes to PP1-mediated dephosphorylation of eIF2α together with its homolog …

A Signal To Divide: Apoptotic Extracellular Vesicles As Carriers Of Mitogenic And Immunogenic Signals, Safia Essien, Safia Essien

Dissertations & Theses (Open Access)

Efficient replacement of dead cells in epithelial tissue is crucial for maintaining barrier function and tissue homeostasis. Apoptotic cells can signal to neighboring cells to stimulate proliferation and compensate for cell loss and maintain overall cell numbers in normal physiology and cancer. While dying cells can transmit instructive cues to neighboring cells, the molecular mechanisms that induce cell division are not well understood. Recent evidence suggests that apoptotic bodies (ABs) or apoptotic extracellular vesicles (AEVs) mediate cell-to-cell communication and carry diverse biologically active cellular cargo which can influence cell proliferation. This dissertation visualizes and characterizes AEVs in larval zebrafish and …

Functional Analysis Of The Gamma-Aminobutyric Acid Type A Receptor Subunit Alpha-1 (Gabra1) Gene During Zebrafish Development., Nayeli Gabriela Reyes-Nava

Open Access Theses & Dissertations

The GABRA1 gene encodes for the alpha-1 (α1) subunit of the Gamma-Aminobutyric acid type A receptor (GABAAR), which are the primary modulators of synaptic inhibition in the central nervous system (CNS). Alpha-1 subunits are essential for maintaining the normal function of native receptors and contribute to over 60% of all GABAARs in the CNS. Remarkably, a broad spectrum of neurodevelopmental and epilepsy-associated disorders have been linked with mutations in the GABRA1 gene. However, the developmental, behavioral, and molecular mechanisms underlying GABRA1-associated epileptic disorders remain to be fully understood. Hence, the overarching goal of this dissertation is to investigate the behavioral …

Correlation Between Stress Coping Styles And Genetic Variation In The Adenosine Pathway Of Zebrafish, Sydney Klucas

Theses/Capstones/Creative Projects

Anxiety is a common mental disorder and is caused in part by dysregulation of neurotransmitter systems. Specifically, the adenosine pathway has been correlated with anxiety disorders, but it is not yet known if genetic differences in this pathway are responsible for anxiety-related behavior. In this study, I used bioinformatic analyses (EdgeR and WGCNA) to investigate an RNA-sequencing dataset to assess differences in differential gene expression and gene co-expression networks within the adenosine pathway of zebrafish with different stress coping styles. These analyses included three brain regions: basolateral amygdala, hippocampus, and habenula. The differential gene expression analysis demonstrated that the adenosine …

Neurogenetic Function Of Host Cell Factor-1, Victoria Lynn Castro

Open Access Theses & Dissertations

Neural precursor cells (NPCs) are the stem-like cells of the developing brain. These cells differentiate into the differentiated cell types of the central nervous system. If disruption occurs in the number of total NPCs formed or their survival, various disorders including learning disabilities, behavioral problems, or epilepsy can occur. This dissertation describes how the HCFC1 gene controls the proliferation and differentiation of NPCs. HCFC1 encodes for a transcriptional co-factor that regulates the growth and metabolism of stem cells, including NPCs. Mutations in HCFC1 cause cblX syndrome, a neural developmental disorder that affects the nervous system and causes microcephaly, epilepsy, and …

Is The Area Postrema Resistant To The Anesthetic Effects Of Ms-222?, Austin T. Csiszar

Cal Poly Humboldt theses and projects

MS-222 is a common fish anesthetic that dampens neural activity by inhibiting neuronal voltage-gated sodium channels (Na_v) required for action potentials. Previous experiments conducted by our lab’s primary investigator using fluorescence calcium imaging in zebrafish neurons resulted in the observation that neurons in the area postrema (AP), a circumventricular organ located in the dorsal hindbrain, displayed prominent fluorescence intensity during MS-222 induced sedation. I used the genetically encoded calcium indicator GCaMP6s to conduct fluorescence calcium imaging with the goal of recording neural activity in the larval zebrafish area postrema and optic tectum to compare the anesthetic effects of …

Knockout Of Endospanin 1 Via Crispr In Zebrafish, Danio Rerio, Jared Kittinger

Williams Honors College, Honors Research Projects

I made endospanin 1 knockout (KO) zebrafish to examine its effects on lipid and bone metabolism. Endospanin 1, or leptin receptor overlapping transcript (leprot), is a cytosolic protein linked to the protein hormone leptin that influences the trafficking of leptin receptors to the plasma membrane of cells. Genes for endospanin and tyrosinase (a pigmentation enzyme) were targeted via a microinjection of guide RNAs and CRISPR Cas9 into zebrafish embryos at 2-4 cell stages. I was able to disrupt the endospanin 1 gene (based upon the disruption of tyrosinase), but very few mutant zebrafish fully developed into adults. Only low KO …

The Effects Of Sex On Zebrafish Bone Metabolism, Simon Bagatto

Williams Honors College, Honors Research Projects

I investigated the effects of diet and sex on zebrafish bone metabolism. Zebrafish were subjected to either a high-calorie or low-calorie diet over a five-week period. After this diet, zebrafish scales were used to measure alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP) enzyme activity with a fluorescent substrate. The enzyme activities were markers of the osteoblasts (ALP) and the osteoclasts (TRAP). These data were compared among sex and diet of the zebrafish using a repeated measures ANOVA statistical test. I also measured the number of resorption lacunae per scale (another indicator of osteoclast activity). The results showed higher TRAP …

The Impact Of Polychlorinated Biphenyls On The Development Of Zebrafish (Danio Rerio), Megan Moma

WWU Graduate School Collection

Polychlorinated biphenyls (PCBs) are a group of 209 highly stable molecules that were used extensively in industry. Although their commercial use ceased in 1979, they are still present in many aquatic ecosystems due to improper disposal, oceanic currents, atmospheric deposition, and hydrophobic nature. PCBs pose a significant and ongoing threat to the development and sustainability of aquatic organisms. Our hypothesis is that PCB concentration will significantly affect development. Zebrafish (Danio rerio) were exposed to a standard PCB mixture (Aroclor 1254) for the first 5 days post fertilization, as there is a gap in knowledge during this important developmental …

The Effects Of Cucurbitacin B And Silmitasertib On Metastasis And Itga6 Using The Zebrafish Tumor-Xenograft, Alexandra Griffis

Electronic Theses and Dissertations

Ninety percent of cancer deaths are resultant from the metastasis of cancer cells. When cancer cells translocate through blood vessels or the lymphatic system, they may form tumors outside of their primary site. The processes of metastasis can begin quickly; after onset, metastasis is unforgiving, as it does not participate with the body’s physiological systems in an orderly way. In the past, our lab produced results indicating that a cell adhesion molecule, Integrin Alpha-6, may contribute to cancer cells' ability to metastasize. Integrins mediate interactions between the cell and the Extracellular Matrix (ECM), regulating cell attachment and cell migration. With …

The Effects Of Diatom-Specific Polyunsaturated Aldehydes On Larval Zebrafish (Danio Rerio), Rachel Raymer

WWU Graduate School Collection

Diatoms are ubiquitous in marine planktonic and benthic environments and are common in diets for many lower-trophic organisms. Certain species of diatoms produce polyunsaturated aldehydes (PUAs) that can exist in particulate and dissolved forms. Diatom PUAs are known for negatively affecting the fecundity of their primary consumers, including invertebrate grazers like copepods and echinoderms. However, little is known about the effects of diatom PUAs on vertebrates that may be exposed to dissolved or ingested PUAs due to overlapping distribution with diatom populations. The purpose of this study was to test whether dissolved diatom PUAs affect the early life stages of …

Examining The Effects Of Hyperglycemia On Retinal Regeneration, Emmanuella Ec Kyllians

Theses and Dissertations--Biology

Diabetes is a disease that affects how the body processes glucose, leading to a cascade of other complications, one of which is diabetic retinopathy (DR). DR is a neurovascular disease that affects the retina and research has revealed the role of retinal neurodegeneration in its development. Animal models like zebrafish have emerged as important tools for studying the molecular and cellular mechanisms underlying DR. On the other hand, retinitis pigmentosa (RP) is a degenerative disease that involves the progressive degeneration of rods and cones, and hyperglycemia has been found to impair regeneration. Therefore, this study aims to examine the effects …

Developmental Exposures To Pfas Mixtures Impair Elongation Of The Exocrine Pancreas In Zebrafish (Danio Rerio), Emily M. Formato

Masters Theses

Poly- and perfluoroalkyl substances (PFAS) are a class of bioaccumulative toxicants used in numerous industrial and commercial products. Perfluorooctanesulfonic acid (PFOS) alters pancreatic organogenesis during development, and perfluorohexanesulfonic acid (PFHxS) has been suggested as a replacement for PFOS due to its shorter carbon chain, but they are often found together in surfactants, such as legacy aqueous film-forming foam. This study investigates how developmental exposures to a PFAS mixture (PFHxS + PFOS) impact the developing exocrine pancreas. Zebrafish embryos (Tg(ptf1a:GFP)) were exposed to 0.01% DMSO, or 8, 16, 32 μM PFHxS alone, 16 μM PFOS alone, and 8, 16, and 32 …

Mapping Auditory And Vestibular Response Neurons In The Optic Tectum Of Larval Zebrafish, Erika Marks, Arminda Suli

Undergraduate Honors Theses

Cellular mapping of the sensory receptive fields of brain structures is a critical step in understanding function and dysfunction in development. Deficits in the ability to receive, process and integrate the senses leads to difficulties recognizing and responding appropriately to stimuli. To understand how the senses are integrated, it is first necessary to map the neurons receptive to inputs from individual senses. The optic tectum of zebrafish is a structure known for its homology to the superior colliculus of mammals and is a center of multisensory integration – including visual, auditory, and somatosensory stimuli. However, the microcircuitry of this important …

Effects Of Exercise On Fear Behavior In Zebrafish, Tyler Kuni

Undergraduate Theses and Capstone Projects

The experiment focuses on the behaviors of zebrafish when they are exposed to an environment that causes anxiety or fear behavior and how controlled amounts of exercise will affect the behaviors of the fish. Fearful behavior was measured by the activity level and area of the swimming during the fear conditioning process. We investigated whether the changes in activity level, areas of swimming in the tank, and depth of swimming in the tank were significant as well as if there was an interaction effect between the fear conditioning and our dependent variables. Zebrafish were randomly assigned to three groups: the …

Characterizing The Roles Of Cannabinoid Receptor 1 & 2 In Zebrafish Behavior, Metabolism, And Seizure-Induced Activity, Kayci Kimmons

Honors Theses

Epileptic disorders like Dravet Syndrome require novel studies to determine the most ideal treatment. New research linking the endocannabinoid system (ECS) to epileptic disorders is arising, but there is still much to be discovered about the function and regulatory impact of the endocannabinoid system and its receptors in epilepsies like Dravet. In this study, knockout models of larval and adult zebrafish (Danio rerio) were used to investigate the roles of cannabinoid receptors 1 & 2 in behavior, brain mitochondrial metabolism, and seizure-induced activity following exposure to THC and CBD. Larval zebrafish which lacked cannabinoid receptor 1 exhibited increased …

Modeling Developmental, Molecular, And Behavioral Effects Of An Apolipoprotein-E4 Fragment On The Embryogenesis Of Zebrafish, Madyson Mccarthy

Boise State University Theses and Dissertations

Although the increased risk of developing sporadic Alzheimer’s disease (AD) associated with the inheritance of the apolipoprotein E4 (APOE4) allele is well characterized, the molecular underpinnings of how ApoE4 imparts risk remains unknown. Enhanced proteolysis of the ApoE4 protein with a toxic-gain of function has been suggested and a 17 kDa amino-terminal ApoE4 fragment (nApoE4_1-151) has been identified in post-mortem human AD frontal cortex sections. Recently, we demonstrated in vitro, exogenous treatment of nApoE4_1-151 in BV2 microglial cells leads to uptake, trafficking to the nucleus and increased expression of genes associated with cell toxicity …

Assessing The Effects Of Levonorgestrel (Lng) On Neural Differentiation In The Developing Hypothalamus Using The Zebrafish (Danio Rerio) As A Model, Gabriella Hinojosa

Theses and Dissertations

Neurogenesis is a critical component of fetal neurodevelopment consisting of: migration, proliferation and differentiation. Health concerns from anxiety to reproductive issues have been linked to alterations of neurogenesis processes. Endocrine disrupting chemicals (EDCs) can mimic hormones vital to this process, potentially adversely affecting the susceptible fetal brain. One EDC, Levonorgestrel (LNG), is a testosterone-like progestin used in birth control methods that has a high bioavailability and long half-life. Using zebrafish embryos as a model, this lab previously discovered that an environmentally relevant LNG dose (5 ng/L) significantly increased anxiety behaviors. The hypothalamus is responsible for the development …

The Role Of The Basement Membrane In Basal Tissue Folding, Elizabeth Falat

Theses and Dissertations

The ability of organs and tissues to function properly is reliant on the correct formation of shape. Proper morphogenesis is carefully regulated by both intracellular dynamics and extracellular mechanical forces. Using the highly conserved zebrafish midbrain-hindbrain boundary (MHB), we investigated the role of the basement membrane, a specialized extracellular matrix that basally lines the tissue, in mediating cell shapes and mechanical tissue stresses during basal tissue folding. A primary component of the basement membrane is laminin-111, a heterotrimer composed of the alpha1, beta1 and gamma1 chains, encoded by lama1, lamb1a, and lamc1 genes respectively. We investigated the role of each …

Spatiotemporal Characterization Of The Prr12 Paralogues In Zebrafish, Renee Jeannine Resendes

Electronic Thesis and Dissertation Repository

Pathogenic variants in the human PRR12 (Proline Rich 12) gene are associated with PRR12-related Neuroocular Syndrome. However, little is known about the gene/protein function. The zebrafish was utilized to address this, as its attributes place it as a premier model to study genes involved in human development and disease. In situ hybridization and RT-PCR of embryos and larvae, and qRT-PCR of adult tissues revealed the spatial and temporal distributions of the prr12 paralogues: prr12a and prr12b. Both paralogues were detected from the maternal and zygotic transcriptomes in a global and diffuse expression pattern, and there was enrichment …

Immunohistochemistry Of The Zebrafish Retina, Alison Ruf

Williams Honors College, Honors Research Projects

The retina is made up of many different essential parts that work together to allow for vision of many different species. Some of these crucial cells and proteins include ganglion cells, bipolar cells, photoreceptors, synaptic markers, such as VGLUT, and receptors that bind glutamate. The basis of this project is to use immunohistochemistry to identify these important aspects of the retina in zebrafish via monoclonal and polyclonal antibodies. The goal being to compare specific cells in a control group retina to an experimental group of zebrafish.

Investigating The Role Of Chd7 And Sox11 In Retinal Cell Development And The Ocular Complications Of Charge Syndrome, Laura Krueger

Theses and Dissertations--Biology

Proper formation of the visual system requires the precise interaction of several embryonic cell lineages, including the neuroectoderm (forms the retina and retinal pigment epithelium), surface ectoderm (forms the lens), mesoderm and cranial neural crest cells (form the ocular blood vessels and anterior ocular structures). When this process is disrupted structural birth defects such as coloboma result, leading to pediatric visual deficits. Ocular developmental defects are often present in larger syndromic disorders. One example is CHARGE syndrome, a genetic disorder characterized by coloboma, heart defects, choanal atresia, growth retardation, genital abnormalities, and ear abnormalities. Pathogenic variants in CHD7 have been …

Examining Development And Function Of Pretectal Visual Neural Circuits In Gs Homeobox 1 Mutant Zebrafish, Alexandra Rose Schmidt

Graduate Theses, Dissertations, and Problem Reports

Brain development requires a coordinated genetic code to regulate initial cell identity determination, migration, and connectivity, to establish function of neural circuits. Independent neural circuits underlie our ability to produce both complex and innate behavioral responses to sensory stimuli that are often conserved across vertebrate organisms. Sensory processing disruptions are associated with several neurodevelopmental disorders (NDDs). Therefore, gene mutations altering neurodevelopment can lead to changes influencing structure and function of individual neural circuits, causing behavioral deviations in sensory responsiveness. Crucial gene networks that define functional properties of sensory domains are often explored using non-mammalian vertebrate models, such as the zebrafish. …

Does Bisphenol-A, An Estrogen-Like Environmental Toxin, Disrupt Expression Of The Neuronal Chloride Exporter Protein During Early Brain Development?, Mayra S. Mendez

Cal Poly Humboldt theses and projects

Bisphenol A (BPA), a manufactured compound found in consumer products, is known to adversely affect early brain development by disrupting normal estrogen signaling. Recently, BPA was reported to suppress expression of a gene encoding the neuron-specific chloride ion transporter, KCC2. Human and animal studies show suppressing KCC2 can cause neuronal and behavioral hyperactivity. Therefore, some adverse effects of BPA may be due to KCC2 suppression and consequent neuronal hyperactivity. This study aimed to determine whether BPA exposure during brain development alters KCC2 expression. A secondary purpose was to evaluate whether a new transgenic zebrafish line, KCC2:mCitrine,could be used to track …

The Role Of Sox4 In Ocular Morphogenesis And Retinal Differentiation, Rebecca Petersen

Theses and Dissertations--Biology

Visual impairment ranges from mild forms that can be corrected with glasses to more severe cases that result in permanent loss of vision. Microphthalmia, anophthalmia, and coloboma (collectively referred to as MAC) account for 11% of cases of pediatric blindness and are a result of improper ocular morphogenesis. Retinitis Pigmentosa (RP) is a retinal degenerative disease that affects 1 in 3000 people worldwide. It is a progressive disorder that initially begins with loss of vision in low light settings due to rod photoreceptor degeneration but progresses to complete blindness upon loss of cone photoreceptors. Currently, there is no cure for …

Understanding The Effects Of Embryonic Hyperglycemia On Retinal Development And Maintenance, Kayla Titialii-Torres

Theses and Dissertations--Biology

Hundreds of millions of people are affected by diabetes worldwide. Whether they are diagnosed with prediabetes or Type I or II diabetes, there are a variety of mechanisms in the pathogenesis of diabetes. Diabetes is a disease which consists of recurring states of hyperglycemia that can be difficult to manage due to either lack of insulin production or improper utilization of insulin. While these mechanisms of action differ, complications induced by diabetes occur in both poorly regulated Type I and II. Common complications of diabetes include nerve damage, kidney damage, and eye damage. Eye damage specifically is called diabetic retinopathy …

The Heritability Of Mandible Length In The Zebrafish (Danio Rerio), Danielle K. Ringo

WWU Graduate School Collection

Jaw shape often plays a significant role in determining feeding niche. Changes to jaw development can alter feeding mechanics and affect an organism’s ability to acquire and/or process food. Derived patterns of jaw morphogenesis are therefore frequently associated with evolutionary shifts in trophic ecology. Changes in thyroid hormone signaling can have strong developmental effects on many aspects of vertebrate anatomy including the shape of the lower jaws. Zebrafish mutants (opallus^b1071) are hyperthyroid and typically undergo excessive mandible elongation. However, hyperthyroid opallus do not always develop elongated lower jaws. This mutant line was maintained at separate universities for …

Studying The Behavioral Role Of The Kcnn2 Gene In Zebrafish Embryos, Gina Dragonette

Honors Theses

Introduction: Mutations in the KCNN2 gene have been linked to phenotypes of movement disorders and autism. The goal of this experiment was to use the CIUSPR-Cas9 system to create a deletion within the KCNN2 gene and to observe the effect on the expression of movement in zebra fish embryos.

Methods: Linearized Cas9 DNA and guide DNA underwent in vitm transcription (IVT) reactions to produce RNA to be injected into zebrafish embryos at the one cell stage. KCNN guide RNAs were produced for the KCNN2 and KCNN3 genes (to prevent KCNN3 protein function from compensating for loss ofKCNN2's protein function), and …