Cell and Developmental Biology Commons^™

Bax Activation Through The N-Terminal Bh3-Like Domains Of Vdac1 And Vdac2, Autumn A. Peters

Honors Theses

Voltage-dependent anion channels (VDACs) are central to mitochondrial function by mediating metabolite diffusion across the mitochondrial outer membrane (MOM). Two isoforms­­­­­­­­­­­,­­­­­­­­­­­­­­­­­­­­­­­­­­­­ VDACs 1 and 2, are also implicated in apoptosis based on interactions with Bcl-2 family proteins that control this process, including Bax and Bcl-xL. Activation of the pro-apoptotic protein Bax by interaction with ‘BH3-only’ proteins, such as Bim or Bid, induces MOM permeabilization to signal apoptosis, yet accumulating evidence indicates VDACs 1 and 2 may also affect Bax. We discovered that the VDAC N-terminal domains contain high sequence similarity to the conserved BH3 domains within Bcl-2 family members that …

Spectral Characterization Of Colorectal Cancer Using Excitation-Scanning Hyperspectral Imaging, Natalie Hadad

Honors Theses

Colorectal cancer is one of the most common and deadly cancers worldwide, with a lifetime risk of approximately one in twenty-three for men and one in twenty-five for women. Although early detection significantly improves survival, current diagnostic approaches such as tissue biopsy are invasive, reliant on subjective interpretation, have extended turnaround times, and risk missing flat or depressed lesions that are more difficult to detect than raised polyps. These limitations highlight the need for objective, minimally invasive technologies capable of detecting early tissue changes. This study investigates excitation-scanning hyperspectral imaging (Ex-HSI) as a label-free optical approach for identifying colorectal cancer–associated …

Developing A Protocol For Non-Cellular Transplantation Of Donor Mitochondria Into Recipient Cells, Andrew Parton

Honors Theses

Tumor innervation has emerged as a critical feature of cancer progression, with increased nerve density correlating with enhanced aggressiveness, metastatic dissemination, and poor clinical outcome. However, the functional contribution of neurons to tumor biology remains incompletely defined. We recently identified the intercellular transfer of mitochondria from neurons to cancer cells as a mechanism that promotes tumor progression. We therefore hypothesized that disruption of this transfer could attenuate tumor aggressivity. To directly investigate this process, we developed an approach to isolate mitochondria from donor cells and transplant them into recipient cancer cells independently of canonical cell-cell interactions. Transferred mitochondria were tracked …

Discovering The Genetics Underlying Speciation Traits In Heuchera (Coral Bells), Tajinder Singh

Honors Theses

Hybridization is a key evolutionary mechanism for generating biological diversity, ultimately shaping diversification and adaptation of plant lineages in natural and agroecosystems.  Our understanding of the frequency, distribution, and significance of hybridization is rapidly increasing, but the external driving forces that dictate why certain species hybridize, and others do not remain obscure. The physical traits that shape gene flow and trait sharing among species are likely to be among the key factors that directly control hybridization. Gene flow in most plants is mediated by pollinating insects and thus by pollination syndromes, which are suites of floral traits that have evolved …

Cleft Palate And The Coordination Of Wnt Signaling And Pax9 In Murine Palatogenesis, Landon Wyatt

Honors Theses

Cleft palate is a common craniofacial birth defect that arises when the molecular and morphogenetic events guiding secondary palate formation lose coordination during a narrow developmental window. In mice, successful palatogenesis requires the paired palatal shelves to grow vertically, elevate above the tongue, and fuse at the midline; disruption of any of these steps can result in clefting. Two important regulators of this process are canonical Wnt signaling and the transcription factor Pax9, both of which contribute to normal palatal mesenchymal growth and patterning during early development. This paper first examines whether altered Dkk1/Wnt signaling contributes to the Pax9-null palate …

The Impact Of Pp1Β Cancer-Associated Polymorphisms On Pp1Β Protein Abundance And Cell Survival In Humanized Saccharomyces Cerevisiae, Kaylee Martinez

Honors Theses

PP1β is an understudied protein phosphatase that affects the cell’s ability to reproduce. It has been known to be an important part of the cell cycle. Mutations within PP1β have been found to affect that process and influence the development of cancer and disease. In this thesis, different PP1β cancer and disease associated polymorphisms are investigated through the use of humanized yeast. PP1β yeast variants are tested by using Live/Dead cell assay and western blot techniques. Live/Dead analysis was conducted on different strains grown at permissible temperature (room temperature) and non-permissible temperature (37°C) over the course of 6 or 48 …

A Search For Transcriptional Cofactors Of Gcm/Repo Involved In Determining Glia Cell Fate In Drosophila Melanogaster, Nivedha Kosalram

Honors Theses

In Drosophila melanogaster, the mechanism influencing the differentiation of glia and neurons is directed by a key transcription factor known as glial cells missing (gcm). gcm is a crucial regulator of cell fate decision, functioning as a binary switch to direct glial fate in gcm- positive cells and neuronal fate in gcm-negative cells (Jones, et.al, 1995; Hosoya, et.al, 1995; and Vincent, et.al, 1996). In addition to directing gliogenesis, gcm and its homolog gcm2 facilitate the determination of macrophages and tendon cells, highlighting that gcm acts with supplementary factors to induce various signaling pathways in different contexts (Jones, 2005; Johnson, et …

Transcriptional Analysis Of Inflammatory Responses Of Mouse Trophoblast Stem Cells To Interferon Gamma, Arnaz Maryam Tariq

Honors Theses

The blastocyst faces significant immunological challenges during implantation, including heightened inflammation, which can contribute to implantation failure. Trophoblast stem cells (TSCs) and their differentiated forms (TSC-TBs), derived from the trophectoderm (TE) of the blastocyst, play a crucial role in immune modulation. Previous studies suggest that interferon-stimulated genes (ISGs) derived from TSC and TSC-TB provide antiviral protection to embryonic stem cells (ESCs). These findings show indication of a potential antiviral role for placental progenitors in shielding the blastocyst. Additionally, TSCs and TSC-TBs exhibit selectively attenuated immune responses to the proinflammatory cytokine interferon-gamma (IFN-γ) compared to mouse embryonic fibroblasts (MEFs). The attenuated …

Cell Cycle Regulator Delays Pilus Production In Caulobacter Crescentus, Eli Johnson

Honors Theses

Caulobacter crescentus is a model bacterium for its unusual life cycle. This cycle features three distinct phases- the stalked stage, pre-divisional or actively dividing stage, and the swarmer stage, characterized by the production of a flagellum and multiple pili. Pili are necessary for cell attachment to surfaces and their reuptake into the cell can help drive the cell cycle forward. The gene that encodes the pilus subunit, pilA, is regulated by four binding sites for the cell cycle regulator CtrA. Previous research has determined that one site induces expression while two of the four sites inhibit expression. It is unknown …

Fiddler Crab Gut Microbiomes As An Indicator For Eutrophication Due To Waste-Water Contamination In Estuaries, Madison L. Geraci

Honors Theses

Estuaries are unique biogeochemical zones in coastal systems that provide a myriad of ecological functions: providing habitat for critically important fishery species, diverse species of plants and animals, and contributing to nutrient cycling. Estuaries are vulnerable to anthropogenic stressors as they are intersections between marine and terrestrial communities and are often the hotspot of coastal development and activity. Estuarine sediments act as repositories for anthropogenic contaminants, including excess nutrient runoffs and waste-water pollution that can cause states of eutrophication. Microorganisms are sensitive to these environmental changes, presenting a unique opportunity of being useful bioindicators of environmental stress. Isolating microorganisms from …

Understanding The Functional Role Of Lone Star Tick (Amblyomma Americanum) Threonyl-Trna Synthetase In Reproductive Fitness, Anza Ali

Honors Theses

Introduction: Transfer RNA synthetases are a family of enzymes that catalyze the addition of amino acids to their corresponding transfer RNAs (tRNAs) and play an essential role in protein synthesis. Ticks are obligate hematophagous ectoparasites that rely on prolonged feeding periods to complete their life cycle on a vertebrate host. Tick tRNA synthetases help the tick in continuous protein synthesis for efficient blood feeding and reproductive fitness, thus presenting a potential target for developing new tick control strategies. Studies have been conducted assessing the functionality of different tRNA synthetases such as Valine-tRNA synthetase, Asparagine-tRNA synthetase, Lysine-tRNA synthetase, and Leucine-tRNA synthetase …

Does Vdac2 Have A Bh3 Domain?, Lillian Ferkany

Honors Theses

Mitochondrial outer membrane permeabilization (MOMP) by Bax oligomerization triggers apoptosis. BCl-2 family proteins, classified as BH3 only proteins, pro-survival proteins, or pro-apoptotic proteins, control apoptosis partly through their agonist or antagonistic effects on Bax, which are mediated by their conserved BH3 domains. All BH3 domains form an alpha helix containing 5-7 conserved hydrophobic residues, designated H0-H5, and one conserved aspartic acid that drive interaction with Bax and other ‘multi-domain’ BCl-2 members. BH3 agonists induce Bax oligomerization, while BH3 antagonists sequester Bax to prevent MOMP. We discovered that voltage dependent anion channels (VDACs) in the MOM contain a putative BH3-like domain …

Developing A Continuous Cell Line Derived From The Gulf-Coast Tick (Amblyomma Maculatum) Embryos, Sabrina Walia

Honors Theses

Tick cell lines provide an affordable, reproducible, and efficient in vitro experimental system to study tick-pathogen interactions at the cellular level. Before the first tick cell lines were developed, there was no established method to perform in vitro studies with ticks. Tick research at the organismal level is full of difficulties and challenges. An increase in tick-borne infections and the availability of cutting-edge molecular tools have stimulated the research community's interest in using tick cell lines to answer important biological questions related to new and emerging tick-borne diseases. Unfortunately, not all medically relevant tick species have a cell line available. …

Uptake And Localization Of Poly(2-Vinyl-4,4-Dimethyl Azlactone) Modified With Rhodamine- And Coumarin-Based Molecules In Human Embryonic Kidney (Hek293) Cells, Garrett Tassin

Honors Theses

Fluorescence microscopy is a bioimaging technology that utilizes the excitation and emission of fluorophores to identify cellular structures, processes, and interactive events. Natural and synthetic organic dyes are frequently used in fluorescence microscopy techniques for imaging, therapeutic, and biomedical applications. It is also employed in the development of novel organic dyes for innovative methods of drug delivery and labelling. In this study, we investigate the behavior of an organic dye consisting of the post-polymerization modification of poly(2-vinyl-4,4-dimethyl azlactone) (PVDMA) with tetramethyl rhodamine cadaverine (TMR) and a coumarin-based molecule (DBAC). Using cultured HEK293 cells and fluorescence confocal microscopy, we show the …

Physiological, Temporal, And Environmental Determinants Of Forage Fish Caloric Content In The Gulf Of Mexico, Rebecca R. Walsh

Honors Theses

Within the Gulf of Mexico, forage fish species serve as an important link between lower trophic levels and higher trophic levels, supporting economically valuable predator fish, birds, and mammals. Despite the key ecological role of forage fish, fisheries management efforts are often directed elsewhere; as a result, input data for fisheries models is unavailable for many forage fish species. To fill this knowledge gap, we sought to produce species-specific caloric content values for four forage fish species in the Gulf of Mexico: Atlantic croaker (Micropogonias undulatus), white trout (Cynoscion arenarius), bay anchovy (Anchoa mitchilli), …

Role Of Cdx4 And Sp5l In Zebrafish Development, Wesley Tsai

Honors Theses

The Caudal Type Homeobox transcription factors cdx are a family of genes found in vertebrates that regulates body regionalization and anterior-posterior patterning. They are also responsible for regulating axial elongation, but the mechanisms behind this behavior are not known. Previous studies in mouse embryonic stem cells have shown that the cdx genes are necessary for upregulating the gene sp5 which may be linked to axial elongation. Sp5 is a zinc-finger transcription factor belonging to the specificity protein (sp) family. Our group has used in-situ hybridization experiments on zebrafish embryos to show that sp5-like (sp5l) is transcribed within tailbud tissues that …

Analysis Of Wild Rodent Gut Microbiota As A Function Of Exposure To Ticks And Tick-Borne Pathogens, Joshua Pandian

Honors Theses

Due to advances in high-throughput parallel sequencing, researchers have conducted novel studies exploring relationships between microbiome compositions and different aspects of organism health. Some of these studies have shown that the gut microbiome of rodent models has effects on organism health and behavior and that infection with pathogens and the composition of the skin microbiome are linked to changes in gut microbiome composition. While previous studies have shown how vector microbiota impact vector behavior and pathogen transmission, the effect vectors have on reservoir species microbiomes has been a less prominent focus. We were interested in the relationships between tick parasitism, …

The Role Of Cerium(Iii) In Bacterial Growth And The Microbial Transformation Of Aromatic Hydrocarbons, Shruti Sathish

Honors Theses

Biofilms are communities of surface-attached bacterial cells encased in an exopolymeric matrix. In this state, they are more resistant to antimicrobial treatment and can have adverse effects in medical, agricultural, and industrial settings. Whereas, as biocatalysts, biofilms from nonpathogenic bacteria enhance their performance and stability in catalysis. Unfortunately, there are several challenges when using bacteria in organic transformations due to their complex cellular chemistry. Trivalent lanthanide metals were discovered to serve regulatory roles in some bacterial catalytic processes, including those of Pseudomonas putida KT2440 (P. putida), a non-infectious Gram-negative bacterium. The main goal of our research is to use cerium(III) …

A Survey Of Speech-Language Pathologists' Academic Preparation In Craniofacial Anomalies, Francesca Myerski

Honors Theses

Craniofacial anomalies occur in 1 of 750 newborns a year with the three main types being cleft lip, cleft palate, and cleft lip and/or cleft palate. There are six main factors that affect craniofacial anomalies that are focused on in this research, including genetics, environmental factors, medications, diet, health risks, and surgical procedures/surgeons. The research found that there is a lot of information on craniofacial anomalies and speech-language pathologists need to learn about as much as possible. The lack of knowledge and academic preparations speech-language pathologists have in craniofacial anomalies has decreased their awareness in the birth defect and has …

Arabidopsis Shoot Gravitropism Requires The Mediator Coactivator Complex, Gabriella Newton

Honors Theses

Gravitropism is a plant response to gravity in directing the growth of its organs, such as the root and the stem. Gravity is sensed by statocytes, unique cells that contain starch-filled amyloplasts, which trigger gravitropic growth response by modulating the distribution of growth hormone auxin in the elongation zone of the stem and root. Despite the fact that multiple factors have been identified as key regulators of shoot negative gravitropism, how these factors are regulated at the transcriptional level is largely unknown. In this study, we explore the role of transcriptional machinery in regulating shoot gravity sensing and responses by …

Generation Of Baiap3 Expression Construct For Subcellular Localization And Phenotypic Rescue, Ireland Ann Little

Honors Theses

Mast cells secrete a variety of biological mediators crucial for innate and adaptive immunity. One such mediator is Tumor Necrosis Factor (TNF). TNF released from mast cells and other immune cells has been shown to elicit complex signaling pathways in target cells, resulting in the production of pro-inflammatory cytokines and/or immune cell recruitment. At the molecular level, TNF secretion/exocytosis is mediated by soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) and other regulatory proteins that work together to promote the fusion between secretory granules and the plasma membrane. One regulatory protein, Munc13-4, which facilitates the priming of secretory granules, was recently …

Analysis Of Invasion Proteins Mmp2, Mmp9, Adam12, And Adam17 In Glioblastoma U87mg Cells Treated With Anti-Cancer Compound 3,4-Dimethoxybenzaldehyde, Andras Muranyi

Honors Theses

This research tested the effectiveness of novel compound 3,4-dimethoxybenzaldehyde, demonstrated to have anti-cancer properties. U87MG Glioblastoma cells were exposed to the compound at its LC50 concentration, then processed to collect proteins from the cells. Proteins were analyzed via Western blotting for specific protein levels of matrix metalloproteinase 2 and 9 (MMP) and disintegrin and metalloprotease 12 and 17 (ADAM). Previous research indicates these proteins are involved in the invasive properties of glioblastoma cells. Westerns were quantified with ImageJ and compared using a one-way ANOVA. Results indicate the compound has minimal effect upon the expression of MMP2, MMP9, ADAM12, ADAM17 proteins.

Modeling Jadera Haematoloma’S Phenotypic Variation In The Context Of Its Developmental Plasticity, Michael C. Yorsz

Honors Theses

Phenotypic plasticity is the ability of an organism to integrate information from environmental cues to inform the development of its phenotype and remains understudied in biology. Models of plasticity are needed because evolution in the presence of plasticity is poorly understood. Jadera haematoloma, a hemimetabolous true bug, is an excellent animal model of plasticity, exhibiting a non-linear plastic response to juvenile nutrition that biases adult development into groups with differences in flight capability, wing shape, and fecundity. However, there is a lack of literature consensus regarding the range of developmental outcomes in the species. Some publications report the presence …

Microglia Polarize In Response To Transactive Response Dna-Binding Protein-43 (Tdp-43) And Display Partial Recovery After Removal Of The Stimulus, Alicen Wilcox

Honors Theses

The proper response to protein signals is necessary for a healthy central nervous system (CNS), and protein dysregulation is a feature of neurodegenerative diseases. Transactive response DNA-binding protein-43 (TDP-43) is an intranuclear protein, but mislocalization is associated with amyotrophic lateral sclerosis (ALS). TDP-43 is released into the extracellular space where it is sensed by microglia, the CNS-resident immune cells. Our data and the literature suggest that microglia respond to TDP-43 dysregulation by increasing CNS inflammation. The goal of this study was to determine the impact of TDP-43 on microglial function and the extent to which microglia recovery. To study the …

Purification Of Pcbp2 And Its Effect On The Formation Of Dna I-Motifs And Secondary Structures, Nathan Redden

Honors Theses

Several secondary, non-B forms of DNA are associated with the promoter regions of many oncogenes, including the intensely studied c-MYC gene that codes for a transcription factor. Two such secondary structures, G-quadruplexes, and i-motifs, have become topics of interest due to their prevalence within oncogenic promoters, as well as their potential accessibility for drug targeting. B-form DNA can adopt alternative structures under certain conditions. These alternative structures are favored when there is an asymmetric distribution of guanosine and cytosine on complimentary DNA strands. The strand rich in guanosine is able to form a quadruple stranded DNA complex termed a G-quadruplex. …

Molecular Characterization Of Galectin From Amblyomma Americanum In Context Of Α-Gal Syndrome, Sumar Beauti

Honors Theses

The lone star tick Amblyomma americanum is a vector of various disease-causing pathogens and tick-borne alpha-gal syndrome (AGS) with rapidly expanding populations in the south- and northeast regions of the United States. This study aimed to molecularly characterize galectin and determine its involvement in galactose-α-1,3-galactose (α-gal) synthesis, transport, reproductive fitness, and microbial homeostasis in this tick. The lone star tick galectin possesses two conserved carbohydrate recognition domains and shares homology with other Ixodid tick galectins. Time and tissue-dependent expression data shows that galectin is constantly expressed in salivary glands, midgut, and ovary tissues. An RNA interference approach was used to …

Breast Cancer Sub-Clones That Metastasize To Lung And Bone Exhibit Different Metabolic Preferences, Mollie Merrell

Honors Theses

Metastasis is responsible for the majority of cancer related deaths. In breast cancer the lungs and bones are the major sites for metastasis. Previous studies used the metastatic aggressive MDA-MB-231 breast cancer line to isolate sub-clones that preferentially invade the lungs (LM line) or bones (BoM line). While genes associated with the tissue specific metastasis have been identified, it is unknown if metabolic adaptations contribute to the growth of the LM and BoM lines in their respective organs. The goal of this study was to test the hypothesis that the LM and BoM lines exhibit differences in glucose and glutamine …

Exploring The Role Of Ccdc141 In Zebrafish Heart Development, Luci Strong

Honors Theses

Zebrafish are a good model organism in which to identify genes important for heart development because most human genes have a zebrafish ortholog and genes in zebrafish are relatively simple to modify. ccdc141 is a gene with unknown function, which we originally found to be highly expressed in cardiac cells in mouse embryos. In this project, we are seeking to determine the function of ccdc141 using zebrafish. To perform this experiment, we are targeting ccdc141 for mutation using CRISPR/Cas9. In this project we have analyzed ccdc141 to determine regions of conservation, designed CRISPR guides to target those regions (exon 2 …

Evaluation Of The Bax-Vdac Interaction And Their Influence On Apoptosis In Drosophila Melanogaster, Frances Marie Gatlin

Honors Theses

Apoptosis, also known as programmed cell death, is a cellular process used for development or for when cells undergo injury or stress. The Bcl2 family of proteins includes both pro-apoptotic and anti-apoptotic proteins that control the intrinsic pathway of apoptosis. Understanding the mechanisms and influence these proteins have on apoptosis is an important area of research focused on in Dr. Jones’s lab. Evidence shows a homology amongst the Bcl2 family of proteins at the BH3 domain. Dr. Jekabsons' lab has found a potential homology amongst VDAC 1-3 and the Bcl2 family at the BH3 domain.

Specifically, our lab is using …

Elucidating The Functional Location Of Platelet-Derived Growth Factor Receptor Alpha, Creation Of A Review Chapter On The Use Of Zebrafish In Studying Congenital Heart Defects, And Using 3d Printing To Create Laboratory Tools, Jaret Lieberth

Honors Theses

Congenital heart defects (CHDs) are currently the most prevalent form of birth defect in the United States. Their combined frequency and severity make necessary a deeper understanding of the molecular underpinnings guiding heart formation. The first major step in cardiac morphogenesis, cardiac fusion, involves the medial movement of bilateral populations of myocardial precursor cells to the embryonic midline, where they merge to form the primitive heart tube. Although crucial to subsequent organogenesis, the precise mechanisms governing cardiac fusion remain unknown. Previous studies have found that a mutation in platelet-derived growth factor receptor alpha (pdgfra), called refuse-to-fuse (ref), results …