Life Sciences Commons^™

The Development Of Immunohistochemistry (Icc) And Transcriptomic Protocols To Uncover Neurophysiological Markers Of Stress In Red-Eyed Tree Frogs As A Novel Biome, Shirin Dadina

Senior Theses and Projects

Adverse childhood experiences (ACEs) include various forms of physical and emotional abuse that lead to detrimental long term effects. Agalychnis callidryas (red-eyed tree frogs) have been identified as a novel biomedical model for ACEs. Red-eyed tree frogs demonstrate an escape hatch mechanism under stressors including those from predators, flooding and hypoxia where the embryo undergoes an accelerated developmental process similar to what is observed in the human population regarding precocious puberty. In order to assess this model for neurobiological outcomes of ACEs, morphological and genomic protocols must be developed and validated which is this study’s focus. Paraffin embedding and microtome …

Optimization Of Tyrosine Hydroxylase Antibody For Immunohistochemistry Fluorescence Detection In Zebrafish (Danio Rerio), Madison Thurber

Theses/Capstones/Creative Projects

Dopamine is an important neurotransmitter produced through the catecholamine synthesis pathway that affects brain activity. Unregulated dopamine levels can lead to various diseases such as Parkinson’s Disease or attention deficit hyperactivity disorder (ADHD). Optimization of an immunohistochemistry protocol will allow for the quantification of tyrosine hydroxylase antibody, which indirectly allows for dopamine quantification in dopaminergic regions within the brain. However, the antibody concentration to give the optimal signal-to-noise ratio in IHC varies across studies. Through this experiment, I determined the concentration of tyrosine hydroxylase (TyrH) antibody for immunohistochemistry that gave the best signal-to-background noise ratio within several known dopaminergic regions …

Virological And Pathological Investigation Of Canine Parvovirus-2 (Cpv-2) With The Assessment Of The Genetic Variability Of Field Strains, Zeynep Karapinar, Musa Karaman, İhsan Kisadere, Mustafa Usta, Mehmet Özkan Ti̇murkan

Turkish Journal of Veterinary & Animal Sciences

Canine parvovirus-2 (CPV-2), which is common worldwide, is a highly contagious pathogen in domestic dogs and some wild carnivore species. The agent causes a multisystemic infection in affected animals, especially acute hemorrhagic enteritis and fatal myocarditis in puppies. The study aimed to reveal the presence of virus in dogs with clinical symptoms of CPV-2 infection using virological, pathological, and immunohistochemical methods and make molecular characterization based on partial VP-2 gene of strains circulating in the region. As a result of PCR studies, 9 out of 16 rectal swab samples and 17 out of 38 blood samples were positive. A phylogenetic …

Central Activation Of Orl-1 Receptors In Heart Failure Models, Jeffrey Angell

Theses & Dissertations

Heart failure is a prevalent and debilitating disease associated with high morbidity and mortality rates worldwide and is the leading cause of death in the United States. While various therapeutic strategies have been developed and approved to manage heart failure, exploration of novel targets and mechanisms are essential for improving patient outcomes. This thesis investigates the therapeutic potential of the central activation of opioid receptor-like 1 (ORL-1) by nociceptin in a rodent heart failure model.

Furthermore, this study aims to elucidate the central mechanisms underlying nociceptin induced cardiovascular and renal effects in heart failure. This involves investigating neuronal pathways and …

Neural Responses To Magnetic Orientation Information In Songbirds, Madeleine Ir Brodbeck

Electronic Thesis and Dissertation Repository

Effective navigation and orientation is essential for animals to survive. The study of migratory birds provides insights into these processes, particularly through mechanisms like the geomagnetic compass, which uses Earth's magnetic field for directional information. A brain region in songbirds relevant to this is cluster N, known for its potential role in processing geomagnetic information. Notably, cluster N appears exclusive to nocturnally migratory birds, being active solely at night, and lesion studies reveal that an intact cluster N is necessary for geomagnetic compass orientation. However, given the scarcity of empirical data concerning cluster N, substantial questions persist regarding its function. …

Sex Differences In Stress Reactivity, Brain Morphology, And Oxytocin In The Hypothalamus Of The Gray Short-Tailed Opossum (Monodelphis Domestica), Esperanza I. Zacarias, Daniela Rodriguez, Alexandra Chalons, Sasawan Heingraj, Nicole Altamirano, Joseph Rafac, John L. Vandeberg, Mario Gil

Research Colloquium

Understanding the effects of stress on behavior and cognition is important due to its impact on mental health and wellbeing (Schneiderman et al. 2005). Translational animal research can contribute to the development of new treatments that can improve therapeutic outcomes and our understanding of the neurobiology of stress. In the present study, we complement behavioral stress reactivity with immunohistochemical localization of oxytocin in the hypothalamus, a neuropeptide that regulates stress (Neumann & Slattery, 2016). Oxytocin has potential therapeutic use for mental health disorders (Neumann & Slattery, 2016), and the effects of oxytocin seem to be sexually dimorphic (Love, 2018). Using …

Temporal, Phenotypic, And Quantitative Characterization Of Thyroid Infiltrating Mononuclear Cells During Development Of Spontaneous Autoimmune Thyroiditis In Obese Strain Chickens, Katelyn M. Clark

Poultry Science Undergraduate Honors Theses

The Obese strain (OS) of chickens spontaneously develops autoimmune thyroiditis and is a well-established biomedical model for Hashimoto’s thyroiditis in humans. Both conditions are characterized by the infiltration of the thyroid glands with mononuclear cells resulting in destruction of the thyroid tissue and impairment of the thyroid’s endocrinological functions. Past studies have described mononuclear cell infiltration in thyroids of the OS chickens, but the time-course, cell composition, and relative amounts of the various mononuclear cells infiltrating the thyroids has not been well defined. In this project, the mononuclear cell infiltrate was characterized phenotypically and quantitatively in OS chickens over the …

Distribution Of The Misfolded Isoform Of The Prion Protein In Peripheral Tissues And Spinal Cord Of Rocky Mountain Elk (Cervus Elaphus Nelsoni) With Naturally Occurring Chronic Wasting Disease, Terry R. Spraker, Thomas Gidlewski, J. G. Powers, T. A. Nichols, M. A. Wild

USDA Wildlife Services: Staff Publications

Chronic wasting disease (CWD) is an infectious transmissible spongiform encephalopathy of cervids associated with the presence of a misfolded prion protein (PrPCWD). Progression of PrPCWD distribution has been described using immunohistochemistry and histologic changes in a single section of brain stem at the level of the obex resulting in scores from 0 (early) to 10 (terminal) in elk with naturally occurring CWD. Here we describe the spread and distribution of PrPCWD in peripheral tissues and spinal cord in 16 wild and 17 farmed Rocky Mountain elk (Cervus elaphus nelsoni) with naturally occurring CWD and correlate these findings with obex scores. …

Examination Of Canine Herpesvirus-1 In Dogs By Serological, Molecular, And Pathological Methods, Serkan Yildirim, Fatma Gülten Bayraktar, Hakan Aydin, Yavuz Seli̇m Sağlam, Gi̇zem Eser, Muhammet Bahaeddi̇n Dörtbudak

Turkish Journal of Veterinary & Animal Sciences

In this study, it was aimed to examine Canine Herpesvirus-1 (CHV-1) with different diagnostic methods and investigate the pathological changes in positive samples. For this purpose, 100 dog uteruses of different ages and breeds, were removed by ovariohysterectomy and their blood samples were used as the study material. Enzyme-linked immunosorbent assay (ELISA) and Indirect Fluorescent Antibody Test (IFAT) for serological diagnosis, polymerase chain reaction (PCR) method for molecular diagnosis, immunohistochemistry (IHC) and immunofluorescence (IF) staining for immunopathological diagnosis were performed on the samples. Additionally, hematoxylin and eosin (HE) staining was performed to examine the histopathological changes in the CHV- 1-positive …

Purification And Characterization Of An Anti-Basigin Monoclonal Antibody: Application To The Study Of Human Uterine Cells And Tissues, Jeannine M. Barna

All NMU Master's Theses

Between 1990 and 2017, the global rate of female infertility increased by over 15% (Sun et al., 2019). An embryo's failure to implant into the uterus is the primary cause of early pregnancy loss. Several studies have identified molecules necessary for successful implantation, but their function during implantation remains poorly understood (S. Zhang et al., 2013). The cell surface glycoprotein basigin-2 is necessary for embryo implantation in the mouse and plays a role in several cellular functions, including cell communication and extracellular matrix remodeling (Chen et al., 2009; K. Li & Nowak, 2020). In vitro studies using human uterine cells …

The Effect Of Licogliflozin (Sglt1/2 Inhibitor) On Diabetes And Cardiac Complications, Alanoud Gharib Alblooshi

Theses

Diabetes mellitus (DM) is a chronic endocrine disease affecting millions of people worldwide. In spite of the advances made in the management of DM, poor glycemic control and diabetes complications are still very common. There is a continuous search for new and more effective drugs to treat DM. One of the drugs currently in clinical trials for the treatment of DM is licogliflozin (LIK066), a dual SGLT1/2 inhibitor, which can be used to treat obesity and diabetes. LIK066 inhibits glucose reabsorption in the kidney and small intestine, thereby reducing hyperglycemia. This study aims to investigate the efficacy of licogliflozin on …

Visualization Of Eye Development In Annual Killifish Using Whole-Mount Immunohistochemistry, Carmen Z. Rodriguez, Jason Podrabsky

Student Research Symposium

Vertebrate eye development is highly conserved. Current models have identified the gene networks involved in eye development, but lack the context of extreme environmental conditions that challenge these fundamental programs. Typical vertebrates live for many years while annual killifishes can complete their entire adult lifespan in a matter of weeks. Thus, annual killifishes are a strong model for age-related diseases such as age-related macular degeneration (AMD); an irreversible loss of central vision. Annual killifishes enter embryonic diapause; a period of developmental and metabolic dormancy. These embryos are extremely resistant to environmental stresses such as hypoxia/anoxia that would cause irreparable damage …

Characterizing Rat Lateral Amygdala Interneurons By Colocalization Of Calcium-Binding Proteins Via Serial Multiplex Immunohistochemistry, Ethan Gasteyer

Honors Scholar Theses

The lateral amygdala is a brain structure that plays an important role in regulating fear and anxiety. Some anxiety disorders are hypothesized to develop from failures in this local inhibitory circuit. Distinct populations of these inhibitory neurons express patterns of calcium- binding proteins and neuropeptides that suggest differences in functionality within the lateral amygdala. Furthermore, these patterns of expression are compared between male and female rats to identify sex differences in the lateral amygdala. Previous studies have reported sex differences in amygdala activation and connectivity, but very little is known about the sexual dimorphism of calcium-binding protein and neuropeptide expression. …

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.

How Salivary Glands Recover From Fibrotic Injury, Gabriella Majka

Biological Sciences

Extracellular matrix deposition is required for repair after injury, but if left unresolved can result in fibrosis. Fibrosis is an excess deposition of extracellular matrix proteins following injury or aging that leads to eventual organ dysfunction. One surgical model that we have used to study mechanisms of fibrosis is the salivary gland ductal ligation model. During the process of ligation, a clip is placed on the main ducts of the submandibular (SMG) and sublingual gland (SLG) which leads from the salivary glands to the mouth. This causes the gland to atrophy, or waste away, resulting in decreased acinar cell differentiation …

Lipid Droplet Membrane Proteome Remodeling Parallels Ethanol-Induced Hepatic Steatosis And Its Resolution, Carol A. Casey, Terrence M. Donohue Jr., Jacy L. Kubik, Vikas Kumar, Michael J. Naldrett, Nicholas T. Woods, Cole P. Frisbie, Mark A. Mcniven, Paul G. Thomes

Nebraska Center for Biotechnology: Faculty and Staff Publications

Abstract Lipid droplets (LDs) are composed of neutral lipids enclosed in a phospholipid monolayer, which harbors membrane-associated proteins that regulate LD functions. Despite the crucial role of LDs in lipid metabolism, remodeling of LD protein composition in disease contexts, such as steatosis, remains poorly understood. We hypothesized that chronic ethanol consumption, subsequent abstinence from ethanol, or fasting differentially affects the LD membrane proteome content and that these changes influence how LDs interact with other intracellular organelles. Here, male Wistar rats were pair-fed liquid control or ethanol diets for 6 weeks, and then, randomly chosen animals from both groups were either …

Novel Quantification And Localization Of Water And Solute Transporters In The Tissues Of The Spiney Dogfish (Squalus Acanthias), Tolulope B. Ojo

Electronic Theses and Dissertations

The dogfish, Squalus acanthias is a marine cartilaginous elasmobranch found in the North Atlantic and Pacific oceans. Dogfish synthesize and excrete urea as a product of nitrogen metabolism. They also convert ammonia into urea and retain this urea, such that their plasma is isosmotic or slightly hyperosmotic to the surrounding seawater. To facilitate the regulation of body fluid and to maintain ionic concentrations and osmotic pressure, Dogfish use ion and solute transporters (e.g., NCC, UT-1) and aquaporin water channel proteins. Studies have identified some of the aquaporin genes in the elasmobranch genome, but their functions are mostly uncharacterized. Recent transcriptomic …

Immunohistochemical Localization Of Prolactin Receptor (Prlr) To Hodgkin's And Reed-Sternberg Cells Of Hodgkin's Lymphoma, Rajendra Gharbaran, Onyekwere Onwumere, Naomi Codrington, Latchman Somenarian, Stephen Redenti

Publications and Research

Prolactin receptor (PRLR), a type-1 cytokine receptor, is overexpressed in a number of cancer types. It has attracted much attention for putative pro-oncogenic roles, which however, remains controversial in some malignancies. In this study, we reported the localization of PRLR to the Hodgkin's and Reed-Sternberg (HRS) cells of Hodgkin's lymphoma (HL), a neoplasm of predominantly B cell origin. Immunohistochemistry performed on 5-μm thick FFPE sections revealed expression of PRLR in HRS cells. Cellular immunofluorescence experiments showed that the HL-derived cell lines, Hs445, KMH2 and L428 overexpressed PRLR. The PRLR immunofluorescent signal was depleted after treating the cell lines with 10 …

The Long-Term Persistence And Tissue Tropism Of Brazilian Zika Virus In Monodelphis Domestica, Marisol Morales

Theses and Dissertations

Zika virus (ZIKV) rose as a major public health concern due to the congenital abnormalities of infants born to infected mothers during the 2015 Latin American outbreak. While animal models such as transgenic mice and nonhuman primates are used to study ZIKV pathogenesis, they display various limitations. Additionally, the persistence and tissue distribution of long-term ZIKV infection remains unknown. We describe the laboratory opossum, Monodelphis domestica, as a potential new model for ZIKV pathogenesis research to address these issues. We utilized immunohistochemistry, RT-PCR, and ELISA to show that: infant animals inoculated intracerebrally were susceptible to infection, ZIKV infection persisted …

Insights Into Desiccation Tolerance: Properties Of Late Embryogenesis Abundant Proteins From Embryos Of Artemia Franciscana, Blase Matthew Leblanc

LSU Doctoral Dissertations

LEA proteins are a family of intrinsically disordered proteins that are expressed in various life stages of anhydrobiotic organisms and have been strongly associated with survival during water stress. The brine shrimp Artemia franciscana is the only known anhydrobiotic animal that expresses LEA proteins from Groups 1, 3, and 6. Here, I report that AfrLEA6, a novel Group 6 LEA protein, is most highly expressed in embryos during diapause and decreases throughout pre-emergence development. Notably, there is an acute drop in expression upon termination of the diapause state and the titer of AfrLEA6 during diapause is 10-fold lower than values …

The Effects Of Insulin-Like Growth Factor-1 (Igf-1) And Insulin-Like Growth Factor Receptor (Igfr) Regulation On Cognition And Structure Of Astrocytes, Sariya Khan

Honors Theses

Insulin-like growth factor-1 (IGF-1) is a neuroendocrine signaling hormone that plays an integral role in bone and tissue growth and development. Inhibition of this hormone is known to disrupt the chemistry of the brain, resulting in cognitive impairments such as those seen in many common neurodegenerative diseases. While much research has been conducted on neurons and their relation with IGF-1, the role of astrocytes still needs to be explored. Our research investigates how astrocytes are affected as a result of IGF-1 regulation. Preliminary studies in our laboratory established a connection between IGF-1 and glial fibrillary acidic protein (GFAP), and in …

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 …

Investigating Roles For Rna Turnover Processes In Cell Signaling Through Drosophila Melanogaster Genetic Mosaics, Sudiksha Rathan Kumar

Honors Theses

The process of cell signaling is vital in organisms for proper development as it determines the fate and function of cells. This process is highly regulated by myriad interactions between signaling pathway components and gene expression mechanisms. RNA turnover is a type of RNA processing that degrades RNA. It plays an essential role in homeostasis and environmental changes; however, its influence on signaling pathways is currently unknown. In the present study, the effect of RNA turnover processes on cell signaling was analyzed using the genetic mosaics in Drosophila melanogaster. Fly mosaics were created by breeding RNAi lines targeting cellular …

Expression Of Her-2/Neu In Oral Squamous Cell Carcinoma, Sana Mirza, Naila Hadi, Shahid Pervez, Sultan Zeb Khan, Sameer A. Mokeem, Tariq Abduljabbar, Nawwaf Al-Hamoudi, Fahim Vohra

Department of Pathology and Laboratory Medicine

Background: HER-2/neu is a member of the human epidermal growth factor (HER) family of transmembrane tyrosine kinases, which is significantly associated with the pathogenesis of various cancer types. The aim was to evaluate the expression of HER-2/neu in oral squamous cell carcinoma (OSCC) as a potential biomarker to target antigens for specific immunotherapy in OSCC.
Methods: One hundred and forty histologically diagnosed OSCC cases were identified. Four to five-micrometer thick formalin-fixed, paraffin-embedded tumor sections were stained with Haematoxylin and Eosin (H and E). Histological grade was assessed according to WHO/Broders classification, while tumors were staged according to the American Joint …

Validation Of Antibodies For The Immunolocalization Of Germ Cells In Alligator Gonads., Benjamin Austin

Georgia College Student Research Events

Previous studies of alligators from pesticide-contaminated lakes have shown decreased fertility in comparison to less polluted reference populations. Our research is focused on elucidating potential mechanisms of decreased reproductive success in alligators exposed to environmental contaminants during critical developmental periods. The localization and identification of germ cells during development can provide critical information in assessing future reproductive capability. Germ cells are the unique precursors of gametes, more commonly referred to as eggs and sperm. Specific cell types can be localized through immunohistochemistry (IHC), which involves the use of labeled antibodies to identify specific molecules present in the cells of interest. …

An Immunohistochemical Study Of Her2 Expression In Primary Brain Tumors, Mazaher Ramezani, Shadi Siami, Mansour Rezaei, Sedigheh Khazaei, Masoud Sadeghi

BioMedicine

Background: Primary brain tumors (PBTs) include any tumor in the brain whose prognosis is weak because of their histological characteristics. Aim: Herein, this study aimed to assess the HER2 tumor marker frequency in PBTs. Materials and methods: This study was done on the samples of primary brain tumor diagnosis from 2008 to 2015. Results: Out of 107 patients of brain tumor that had a mean age of 40.4 years (61.7% men), the most common location of the tumor was in the supratentorial region (63.85% cases). The prevalence of high-grade astrocytoma (HGA) and low-grade astrocytoma (LGA) at diagnosis was 43.9% …

Development And Sensory Experience Dependent Regulation Of Microglia In Barrel Cortex, Joshua C. Brumberg, John Kalambogias, Chia-Chien Chen, Safraz Khan, Titus Son, Carolyn Headlam, Cindy Lin

Publications and Research

The barrel cortex is within the primary somatosensory cortex of the rodent, and processes signals from the vibrissae. Much focus has been devoted to the function of neurons, more recently, the role of glial cells in the processing of sensory input has gained increasing interest. Microglia are the principal immune cells of the nervous system that survey and regulate the cellular constituents of the dynamic nervous system. We investigated the normal and disrupted development of microglia in barrel cortex by chronically depriving sensory signals via whisker trimming for the animals’ first postnatal month. Using immunohistochemistry to label microglia, we performed …

Investigation Of Even-Skipped, A Developmentally-Regulated Gene Controlling Neural Segmentation In Dragonflies, Kathryn Bangser

Honors Theses

A comprehensive understanding of the genetic mechanisms underlying pattern formation and neurogenesis is necessary in order to trace the evolutionary history of insect embryogenesis.

One of the most important processes of embryogenesis is the organized pattern formation that allows for proper body segmentation and neural development. Proper segmentation, which relies on a series of specific gene expressions, is necessary for the development of an operational nervous system. Even-skipped (eve), one such regulatory gene, has been studied extensively in certain model organisms, and theories regarding the evolution of its functional role could be further elucidated by visualizing its expression …

Investigating Neuroinflammation And Demyelination In The Nervous System Of Twitcher Mice By The Use Of Immunohistochemistry, Irene Wilson

Natural Sciences and Mathematics | Biological Sciences Master's Theses

Neuroinflammation and demyelination are the hallmark lesions of the Twitcher mouse—the model of Krabbe Disease. By analyzing hemibrains and sciatic nerves via immunohistochemistry, we supported the microglial hypothesis that early activation of microglial cells, macrophages, and globoid cells in the nervous system of Twitcher mice results in specific cellular polarization states that may contribute to myelin loss. The influx of activated macrophages seen in both the central and peripheral nervous systems at days 21 and 17, respectively, accounted for this polarization. Using selected M1 and M2 markers, YKL-40, and GPNMB and CD206, respectively, we proved that microglial cells and macrophages …

Further Characterization Of The Skeletal Phenotype In A Hurler Syndrome Mouse Model And The Ethical Treatment Of Children In Medicine, Anna Marie Mcwoods

MSU Graduate Theses

Mucopolysaccharidosis type I (MPS I) is a rare, autosomal recessive disorder caused by the deficiency of the lysosomal enzyme α-L-iduronidase (IDUA). Absence of IDUA results in the accumulation of dermatan and heparin sulfate and ultimately causes multi-system dysfunction. The most severe form of MPS I is Hurlers syndrome, a rapidly progressive disorder that, if left untreated, is fatal. Current treatment options for diagnosed individuals includes hematopoietic stem cell transplantation (HSCT) and enzyme replacement therapy (ERT). These treatments are able to ameliorate the majority of symptoms with the exception of the bone phenotype. This investigation aimed to further characterize the bone …