Life Sciences Commons^™

Towards Understanding The Function Of An Ets-Like Gene In Nematostella Vectensis: Generation Of A Knockout Mutant Line And A Transgenic Reporter Line, Emily Bullock

Biological Sciences Undergraduate Honors Theses

Due to their unique phylogenic position as sister to Bilateria, Cnidaria are often credited with the utility of allowing for reconstruction of ancestral biology based on characteristics shared with bilaterians and other animals. This factor makes investigation into the nervous systems of cnidarians critical in understanding early neural evolution. Wamides, a class of neuropeptides, have been shown to play a regulatory role in life cycle transitions across many different species. The cnidarian specific Wamide neuropeptide, GLWamide, has previously been identified to play an accelerator role in the metamorphic timing of a specific species of sea anemone, Nematostella vectensis. However, …

Complex Dynamics Of Coral Gene Expression Responses To Low Ph Across Species, Veronica Z. Radice, Ana Martinez, Adina Paytan, Donald C. Potts, Daniel J. Barshis

Biological Sciences Faculty Publications

Coral capacity to tolerate low pH affects coral community composition and, ultimately, reef ecosystem function. Low pH submarine discharges (‘Ojo’; Yucatán, México) represent a natural laboratory to study plasticity and acclimatization to low pH in relation to ocean acidification. A previous >2‐year coral transplant experiment to ambient and low pH common garden sites revealed differential survivorship across species and sites, providing a framework to compare mechanistic responses to differential pH exposures. Here, we examined gene expression responses of transplants of three species of reef‐building corals (Porites astreoides, Porites porites and Siderastrea siderea) and their algal endosymbiont communities …

The Impact Of Prenatal Vape Exposure On Weanling Liver Gene Expression, Isabella R. Fieger, Cristina Isabel Pauig, Mindy Yin, Anna Lipinski, Barrett Blevins, Tyler Plake, Lucas Georges, Katelyn Chism, Isaiah Burciaga, Selma Podbicanin, Cynthia Corbitt, Rachel Neal

The Cardinal Edge

The use of E-cigarettes is a fairly recent phenomenon. Vaping is seen as the “healthier” alternative to smoking cigarettes, yet we know little about the developmental toxicity of commercially available vaping products. In the current study, C57Bl/6 mice were exposed to Vuse Alto Golden Tobacco pods (5% nicotine) 4 days before mating and throughout gestation (GD19) for 1 hour/day every day. Offspring birth outcomes were measured with liver tissue collected at weaning. Gross histology and gene expression in the SIRT1-FXR pathway were examined via qPCR analysis with male and female offspring analyzed separately. No differences in gross morphology or cell …

Morphological And Transcriptional Effects Of Crude Oil And Dispersant Exposure On The Marine Sponge Cinachyrella Alloclada, Yvain Desplat, Jacob F. Warner, Emily J. Blake, Nidhi Vijayan, Marie Cuvelier, Patricia Blackwelder, Jose V. Lopez

Biology Faculty Articles

Marine sponges play important roles in benthic ecosystems. More than providing shelter and food to other species, they help maintain water quality by regulating nitrogen and ammonium levels in the water, and bioaccumulate heavy metals. This system, however, is particularly sensitive to sudden environmental changes including catastrophic pollution event such as oil spills. Hundreds of oil platforms are currently actively extracting oil and gas in the Gulf of Mexico. To test the vulnerability of the benthic ecosystems to oil spills, we utilized the Caribbean reef sponge, Cinachyrella alloclada, as a novel experimental indicator. We have exposed organisms to crude …

Proposing An Rna Interference (Rnai)-Based Treatment For Human Immunodeficiency Virus (Hiv) By Analyzing The Post-Transcriptional Gene Targeting Of Sars-Cov-2, Hepatitis C Virus, And A549 Lung Cancer Cells, Arjun Jagdeesh

Undergraduate Research Posters

Human Immunodeficiency Virus (HIV) is a retrovirus that infects CD4+ T cell lymphocytes in humans, leading to the development of Acquired Immunodeficiency Syndrome (AIDS) if left untreated. While current treatment methods, including antiretroviral combination treatments, effectively limit HIV replication, HIV can evade these treatments due to its high mutation rate. Long-term antiretroviral treatment can also be toxic to patients, meaning patients would benefit from a new mechanism of HIV treatment. RNA interference (RNAi) is an antiviral pathway found in mammals, plants, and insects that involves a small-interfering RNA that is incorporated into a protein complex called the RNA-induced Silencing Complex …

Experiencing Short Heat Waves Early In Development Changes Thermal Responsiveness Of Turtle Embryos To Later Heat Waves, Anthony Breitenbach, Rosario A. Marroquín-Flores, Ryan T. Paitz, Rachel M. Bowden

Faculty Publications – Biological Sciences

Although physiological responses to the thermal environment are most frequently investigated using constant temperatures, the incorporation of thermal variability can allow for a more accurate prediction of how thermally sensitive species respond to a rapidly changing climate. In species with temperature-dependent sex determination (TSD), developmental responses to incubation temperature are mediated by several genes involved in gonadal differentiation. Kdm6b and Dmrt1 respond to cool incubation temperatures and are associated with testis development, while Foxl2 and Cyp19A1 respond to warm incubation temperatures and are associated with ovary development. Using fluctuating incubation temperatures, we designed two studies, one investigating how conflicting thermal …

Testing Ssa4::Ade3 Reporters For Mcs Screening, Chloe Ledford, Rebecca Adams Phd

Science University Research Symposium (SURS)

Gene expression is essential to life and occurs through the processes of transcription of mRNA in the nucleus, export of transcripts to the cytoplasm through the nuclear pore complex (NPC), and translation of the mRNA into protein in the cytosol. The budding yeast S. cerevisiae is a eukaryotic model system used to explore the regulation of mRNA export. Transcripts are able to exit the nucleus through interaction with Mex67 which binds mRNA via adaptor proteins and allows crossing through NPCs. However, during heat shock (42°C) known adaptor proteins are rendered dysfunctional thus halting general mRNA export. Under these conditions, specific …

Identification Of Synonymous Genes And Pathways Implicated In Irritable Bowel Disease And Pancreatic Duct Adenocarcinoma, Lavanya Uppala

Theses/Capstones/Creative Projects

Better understanding and genetic characterization of the gut microbiome will allow for the identification of clinically distinct gastrointestinal diseases. Facilitated by high throughput technologies, intestinal flora analyses have elucidated a broad spectrum of neuropsychiatric, immunological, and allergic disorders linked with this organ system. Microbiome research especially has shed light on underlying factors of intestinal disorders. This interplay of environmental bacteria versus host tissue gene expression may have implications for disease pathogenicity and etiological determination. For instance, pancreatic disorders are common symptoms of irritable bowel disease (IBD), which is thought to affect approximately 7% to 21% of the population [1]. However, …

The Central Dogma: Gene Expression, Ayisha Sookdeo

Open Educational Resources

In this lesson plan, students will learn the basic structure and function of DNA and RNA. They will also learn the process of gene expression. Finally, students will learn about the scientific contributor, Ernest Everest Just, and his contributions to the field of Biology.

Periodic Spatial Disturbances Alter The Expression Of Quorum Sensing Virulence Factors In Pseudomonas Aeruginosa, Laura García-Diéguez

All HCAS Student Capstones, Theses, and Dissertations

Pseudomonas aeruginosa is an opportunistic pathogen associated with severe acute and chronic illnesses. Current antibiotic-based approaches fail to effectively treat P. aeruginosa infections due to the effectiveness and robustness of the quorum sensing signaling system (QS). Pathogenic bacteria, such as P. aeruginosa, employ this population density-dependent communication mechanism to confer antimicrobial resistance, propagate infection, and coordinate the expression of virulence factors, through the production and detection of autoinducing signaling molecules (AI). As such, there is a growing interest in developing novel non-antibiotic-based techniques to attenuate the pathogenicity of P. aeruginosa by disrupting the functionality of its QS system. Previous …

Sex And Starvation Influences Latrotoxin Expression In The Brown Widow Spider, Mattie Harris

Honors College Theses

Widow spiders (genus Latrodectus) are well-known for their potent venom. Seven latrotoxin proteins constitute the main components of widow spider venom. The vertebrate specific (α-latrotoxin) and insect specific (α-latroinsectotoxin) latrotoxins have been well-characterized with respect to structure and function. Regulation of latrotoxin gene expression is not well understood but sex and feeding could be factors influencing production. In this study, I used quantitative qPCR to (1) characterize the expression patterns of both the insect and vertebrate specific latrotoxins in male and female brown widow spiders (Latrodectus geometricus) to characterize sex-biased expression and to (2) study expression patterns …

From Gene Expression To Physiology: A Study Of Chronic Thermal Tolerance In The Mytilus Edulis Species Complex, Lindsey Cate Schwartz

Theses and Dissertations

Rising ocean temperatures are a severe and ever-present threat to marine life. With environmental temperature having such a large impact on organismal performance, understanding the mechanisms which contribute to the ability to survive at higher temperatures is a crucial research focus. Significant progress has been made in discovering these mechanisms on the cellular, biochemical, and physiological levels, but is it much less common for those to be examined together. Using the Mytilus edulisspecies complex as a model system, this dissertation takes a closer look at how prolonged exposure to sub-lethal high temperatures impacts marine organisms on multiple levels of …

Genome-Wide Identification Of Novel Srnas In Streptococcus Mutans, Madeline Claire Krieger, Justin Merritt, Rahul Raghavan

Biology Faculty Publications and Presentations

Streptococcus mutans is a major pathobiont involved in the development of dental caries. Its ability to utilize numerous sugars and to effectively respond to environmental stress promotes S. mutans proliferation in oral biofilms. Because of their quick action and low energetic cost, non-coding small RNAs (sRNAs) represent an ideal mode of gene regulation in stress response networks, yet their roles in oral pathogens have remained largely unexplored. We identified 15 novel sRNAs in S. mutans and show that they respond to four stress-inducing conditions commonly encountered by the pathogen in human mouth: sugar-phosphate stress, hydrogen peroxide exposure, high temperature, and …

Beyond Cortisol: Indicators Of Stress And Negative Feedback In Plasma And Blubber Of Marine Mammals, Jessica Avalos

University of the Pacific Theses and Dissertations

Marine mammals play an important role in ecosystem stability. However, anthropogenic activity is compounding pressure on many already vulnerable populations. A potential consequence of anthropogenic disturbance is physiological stress, which can impact metabolism, immunity, and reproduction, especially if it occurs repeatedly. Previous studies on marine mammals have focused on acute stress, but the impacts of repeated stress are poorly understood. Due to its accessibility on land during haul-outs, the northern elephant seal (Mirounga angustirostris) is a good system in which to study the effects of stress in marine mammals. Stress stimulates the release of glucocorticoid hormones, primarily cortisol. Elevated cortisol …

Cupid’S Arrow: A Tale Of A Complex And Dynamic Protein, Nadine Tietz

WWU Honors College Senior Projects

My capstone project sought to investigate the degree to which ARO4 gene expression is influenced by local vs distant genetic variants. This involved engineering allele-swap strains of ARO4 tagged with a co-expressing fluorescent protein. I then compared protein expression between each strain's genotype which provided an estimate of the proportion of the overall phenotypic variation that can be explained by genetic variation between the underlying genomes.

From my experiment I was able to conclude that ARO4 protein expression was complex and dynamic as both strain background and allele had genetic influences on ARO4 protein expression. In addition, not only did …

Effects Of 17Α-Ethinylestradiol (Ee2) On Gonadal Development And Gene Expression In Larval Mummichog (Fundulus Heteroclitus), Carly Sing-Judge

Theses and Dissertations (Comprehensive)

The mummichog (Fundulus heteroclitus) is a small-bodied estuarine fish that inhabits the North American east coast and is often used as a model adult saltwater fish in environmental bioassays. This study aimed to describe gonadal development and gene expression levels in five-week-old mummichog under control conditions and following 17α-ethinylestradiol (EE₂) treatment in order to better understand the susceptibility of sex determination (SD) and gonadal differentiation (GD) processes following contaminant exposure. In the first experiment, eggs were fertilized, collected and incubated in the laboratory for three weeks, then yolk-sac larvae were grown out for five weeks and …

Assesment Of Antibiotic Resistant Gene Expression In Clinical Isolates Of Pseudomonas Aeruginosa, Dustin Esmond

Biology Theses

Increasing prevalence of nosocomial infections by antimicrobial resistant pathogens resulting in higher mortality rates and financial burden is of great concern. Pseudomonas aeruginosa represents one of six highly virulent “ESKAPE” pathogens that exhibit considerable intrinsic drug resistance as well as mechanisms for acquiring further resistance. As many of these mechanisms are regulated through gene expression, we sought to identify regulatory strategies and patterns at play in 23 clinical isolates collected from Baku, Azerbaijan and Tyler, Texas, USA. Real-time quantitative polymerase chain reaction was performed on six gene targets implicated in resistance and contrasted with antibiotic phenotypes. We found AmpC cephalosporinase …

High-Spatial-Resolution Transcriptomic Map Of The Mouse Lymph Node Microenvironment Using Deterministic Barcoding, Archibald Enninful, Yang Liu, Rong Fan

The Yale Undergraduate Research Journal

Spatial transcriptomics is an emerging approach which characterizes gene expression profiles for a more nuanced understanding of biological processes at a tissue level. This offers significant advantages over traditional omics which require the digestion of tissues and subsequent isolation of cells, during which the spatial information is completely lost. Lymph nodes are an integral part of the immune system and an in-depth analysis of its spatial organization will provide useful insights which can be applicable in the development of novel immunotherapies. In this study, the mouse lymph node is characterized using the newly developed microfluidic-based approach, Deterministic Barcoding in Tissue …

Elucidating The Evolution And Function Of Srnas That Facilitate Bacterial Stress Tolerance, Madeline Claire Krieger

Dissertations and Theses

Non-coding small RNAs (sRNAs) are ubiquitous post-transcriptional regulators found in both gram-negative and gram-positive bacteria. Despite their integral role in many regulatory pathways, including those that facilitate stress-tolerance, little is known about the evolutionary forces that drive the emergence of novel sRNAs, how prevalent sRNAs are within bacterial species, or the functions of a majority of these transcripts. In this work, I first describe the evolution of OxyS, a well-characterized sRNA in gram-negative Enterobacteriaceae that is involved in coordinating the bacterial response to oxidative stress. Next, I identify novel sRNAs differentially produced by the gram-positive oral pathogen Streptococcus mutans in …

The Brain Transcriptome Of The Wolf Spider, Schizocosa Ocreata, Daniel Stribling, Peter L. Chang, Justin E. Dalton, Christopher A. Conow, Malcolm Rosenthal, Eileen Hebets, Rita M. Graze, Michelle N. Arbeitman

School of Biological Sciences: Faculty Publications

Objectives: Arachnids have fascinating and unique biology, particularly for questions on sex differences and behavior, creating the potential for development of powerful emerging models in this group. Recent advances in genomic techniques have paved the way for a significant increase in the breadth of genomic studies in non-model organisms. One growing area of research is comparative transcriptomics. When phylogenetic relationships to model organisms are known, comparative genomic studies provide context for analysis of homologous genes and pathways. The goal of this study was to lay the groundwork for comparative transcriptomics of sex differences in the brain of wolf spiders, a …

The Role Of Med31 And Med12 In Directing Adipogenesis Of Human Adult-Derived Mesenchymal Stem Cells, Joseph Straub

Master's Theses

Selective gene expression is crucial in maintaining the self-renewing and multipotent properties of stem cells. Mediator is a large, evolutionarily conserved, multisubunit protein complex that modulates gene expression by relaying signals from cell type-specific transcription factors to RNA polymerase II. In humans, this complex consists of 30 subunits arranged in four modules: head, middle, tail, and kinase. In our introduction, we show the state of the field of Mediator study with a focus on the critical kinase module. In the following chapters, we used siRNA knockdowns to investigate the roles of the highly-conserved core subunit MED31 and the kinase module …

Differential Gene Expression In Starvation Selected Drosophila Melanogaster, Katherine Schultz

UNLV Theses, Dissertations, Professional Papers, and Capstones

Fruit flies that have been selected for starvation resistance can be used in experimental studies to study physiological processes. Both their genomes and their phenotypes, especially metabolism, change significantly as a result of artificial selection. Starvation selected flies develop slower and are larger due to higher lipid content as adults. We used RNAseq to study gene expression in third instar larval fat body tissue from selected and control lines at the same chronological time point, when the fed control (“FC”) lines were wandering, and the starvation selected (“SS”) lines were in the third larval instar but not yet wandering due …

The Role Of Med31 And Med12 In Directing Adipogenesis Of Human Adult-Derived Mesenchymal Stem Cells, Joseph Straub

Doctoral Dissertations

Selective gene expression is crucial in maintaining the self-renewing and multipotent properties of stem cells. Mediator is a large, evolutionarily conserved, multisubunit protein complex that modulates gene expression by relaying signals from cell type-specific transcription factors to RNA polymerase II. In humans, this complex consists of 30 subunits arranged in four modules: head, middle, tail, and kinase. In our introduction, we show the state of the field of Mediator study with a focus on the critical kinase module. In the following chapters, we used siRNA knockdowns to investigate the roles of the highly-conserved core subunit MED31 and the kinase module …

Transcriptome Prediction Performance Across Machine Learning Models And Diverse Ancestries, Paul Chukwuebuka Okoro, Ryan Schubert, Xiuqing Guo, W. Craig Johnson, Jerome I. Rotter, Ina Hoeschele, Yongmei Liu, Hae Kyung Im, Amy Luke, Lara R. Dugas, Heather Wheeler

Biology: Faculty Publications and Other Works

Transcriptome prediction methods such as PrediXcan and FUSION have become popular in complex trait mapping. Most transcriptome prediction models have been trained in European populations using methods that make parametric linear assumptions like the elastic net (EN). To potentially further optimize imputation performance of gene expression across global populations, we built transcriptome prediction models using both linear and non-linear machine learning (ML) algorithms and evaluated their performance in comparison to EN. We trained models using genotype and blood monocyte transcriptome data from the Multi-Ethnic Study of Atherosclerosis (MESA) comprising individuals of African, Hispanic, and European ancestries and tested them using …

Gene Expression Associated With Disease Resistance And Long-Term Growth In A Reef-Building Coral, Emma R. Kelley, Robin S. Sleith, Mikhail V. Matz, Rachel M. Wright

Biological Sciences: Faculty Publications

Rampant coral disease, exacerbated by climate change and other anthropogenic stressors, threatens reefs worldwide, especially in the Caribbean. Physically isolated yet genetically connected reefs such as Flower Garden Banks National Marine Sanctuary (FGBNMS) may serve as potential refugia for degraded Caribbean reefs. However, little is known about the mechanisms and trade-offs of pathogen resistance in reef-building corals. Here, we measure pathogen resistance in Montastraea cavernosa from FGBNMS. We identified individual colonies that demonstrated resistance or susceptibility to Vibrio spp. in a controlled laboratory environment. Long-term growth patterns suggest no trade-off between disease resistance and calcification. Predictive (pre-exposure) gene expression highlights …

Rna In Situ Hybridization For Detecting Gene Expression Patterns In The Abdomens And Wings Of Drosophila Species, Mujeeb Olushola Shittu, Tessa Steenwinkel, William Dion, Nathan Ostlund, Komal Raja, Thomas Werner

Michigan Tech Publications

RNA in situ hybridization (ISH) is used to visualize spatio-temporal gene expression patterns with broad applications in biology and biomedicine. Here we provide a protocol for mRNA ISH in developing pupal wings and abdomens for model and non-model Drosophila species. We describe best practices in pupal staging, tissue preparation, probe design and synthesis, imaging of gene expression patterns, and image-editing techniques. This protocol has been successfully used to investigate the roles of genes underlying the evolution of novel color patterns in non-model Drosophila species.

Modulation Of Bacterial Fitness And Virulence Through Antisense Rnas, Jess A. Millar, Rahul Raghavan

Biology Faculty Publications and Presentations

Regulatory RNAs contribute to gene expression control in bacteria. Antisense RNAs (asRNA) are a class of regulatory RNAs that are transcribed from opposite strands of their target genes. Typically, these untranslated transcripts bind to cognate mRNAs and rapidly regulate gene expression at the post-transcriptional level. In this article, we review asRNAs that modulate bacterial fitness and increase virulence. We chose examples that underscore the variety observed in nature including, plasmid- and chromosome-encoded asRNAs, a riboswitch-regulated as RNA, and as RNAs that require other RNAs or RNA binding proteins for stability and activity. We explore how as RNAs improve bacterial fitness …

Intragenic Dna Methylation Regulates Insect Gene Expression And Reproduction Through The Mbd/Tip60 Complex, Guanfeng Xu, Hao Lyu, Yangqin Yi, Yuling Peng, Qili Feng, Qisheng Song, Chengcheng Gong, Xuezhen Peng, Subba Reddy Palli, Sichun Zheng

Entomology Faculty Publications

DNA methylation is an important epigenetic modification. However, the regulations and functions of insect intragenic DNA methylation remain unknown. Here, we demonstrate that a regulatory mechanism involving intragenic DNA methylation controls ovarian and embryonic developmental processes in Bombyx mori. In B. mori, DNA methylation is found near the transcription start site (TSS) of ovarian genes. By promoter activity analysis, we observed that 5′ UTR methylation enhances gene expression. Moreover, methyl-DNA-binding domain protein 2/3 (MBD2/3) binds to the intragenic methyl-CpG fragment and recruits acetyltransferase Tip60 to promote histone H3K27 acetylation and gene expression. Additionally, genome-wide analyses showed that the peak …

Fast And Pervasive Transcriptomic Resilience And Acclimation Of Extremely Heat-Tolerant Coral Holobionts From The Northern Red Sea, Romain Savary, Daniel J. Barshis, Christian R. Voolstra, Anny Cárdenas, Nicolas R. Evensen, Guilhem Banc-Prandi, Maoz Fine, Anders Meiborn

Biological Sciences Faculty Publications

Corals from the northern Red Sea and Gulf of Aqaba exhibit extreme thermal tolerance. To examine the underlying gene expression dynamics, we exposed Stylophora pistillata from the Gulf of Aqaba to short-term (hours) and long-term (weeks) heat stress with peak seawater temperatures ranging from their maximum monthly mean of 27 °C (baseline) to 29.5 °C, 32 °C, and 34.5 °C. Corals were sampled at the end of the heat stress as well as after a recovery period at baseline temperature. Changes in coral host and symbiotic algal gene expression were determined via RNA-sequencing (RNA-Seq). Shifts in coral microbiome composition were …

Disruption Of Cellular P-Bodies During An Adenovirus Infection​, La`Quita Randolph, Kevin Neubrecht, Kasey A. Karen

Graduate Research Showcase

Adenovirus has made major contributions in medicine by serving as a model DNA virus to study other viruses, such as human papillomavirus (HPV). Adenoviruses are a diverse family of nonenveloped, double-stranded DNA viruses that are ubiquitous to animals and humans. There are over 67 serotypes of human adenoviruses that can cause a variety of illnesses including, gastroenteritis, conjunctivitis and respiratory infections. Adenovirus can cause these infections by invading host cells and producing an environment that is favorable for viral replication. During the early phases of infection, adenovirus expresses various viral proteins such as E4 11k, which has multiple functions. One …