Course-Directed Research

The Biology Department offers a variety of Research-Intensive courses each semester. In these courses, student learn about the field of research in which they are engaged and work independently or in small teams to design and conduct a research project.

The following courses were offered in Fall 2023 and have students presenting their work in this symposium.

BIOL 322 Animal Ecology, Dr. Brad Lamphere
Animal Ecology studies how animals interact with their world and one another.  Our lab exercises collect data on five topics ranging from ant foraging strategy to fish population estimation to invertebrate community diversity. For the lab final, students use one dataset to analyze and answer an original scientific question. 

The following are the projects presented in this symposium:

  • Poster 1: How Gear Selection Influences Fallfish and Bluegill Catch in Alum Spring Park
    Taylor Nicholson

    Accurately assessing the true population of a species within an environment, particularly in expansive and mobile ecosystems, poses a considerable challenge. To estimate population size and its characteristics, a representative portion of the population must be sampled. Various methods, such as seining or electrofishing, are employed to capture individuals from aquatic populations and examine population dynamics. In Alum Spring Park, Fredericksburg, Virginia, both Lepomis macrochirus (bluegill) and Semotilus corporalis (fallfish) inhabit the same stream but occupy distinct habitats. My investigation aims to explore the potential influence of gear selection on the length of individuals captured, specifically focusing on fallfish and bluegill. The survey involved using seining nets to sample an approximately 85 meter stretch of the stream, followed by a survey using a backpack electrofisher along the same course. I anticipate that electrofishing will yield a larger average fish length for both species compared to seining. This assumption is based on the idea that electrofishing has the capacity to stun larger fish, while larger, more robust fish may evade seining due to their strength and adaptive behavior. Additionally, fallfish are inclined to favor pools, whereas bluegill tends to seek shelter in rocky or vegetated areas, resulting in distinct preferences that affect each species’ catchability which could be influenced by length. This investigation is critical in ensuring a comprehensive representation of the population, particularly considering the varying body types and habitat preferences among these fish species. 
  • Poster 9: The Influence of Food Availability on Dietary Preferences in Little Black Ants and Carpenter Ants
    Anias Saunders

    Carpenter ants and Little black ants are two ants species that are native to Fredericksburg. Ant species’ trophic niches are mostly determined by their dietary preferences, which also have an impact on how these animals interact with one another and the general organization of ant communities. Ecologists can learn more about the competitive relationships, resource distribution, and ecological roles of various ant species within an ecosystem by analyzing food preferences. In this experiment, I investigate what are the nutritional preferences between little black ants and carpenter ants. They were given the choice of a cookie, jam, and tuna over a period of 45 minutes. Each ant was recorded in 5 minute intervals at each food station. Carpenter ants were found more with the cookie and tuna while the little black ants were found at the cookie and jam. These results suggest carpenter ants being carnivores prefer a source of food with some protein whereas the little black ants prefer sugar in their food. 
  • Poster 13: Rural vs. Urban Species Abundance and Richness in Eastern Virginia
    Rachael Harrington and Meghan Ponton

    With the spread of urbanization, understanding how species navigate and utilize resources is necessary to identify the impacts of urbanization on these locations. This study looks into the influence of urbanization on species abundance and richness in urban and rural areas in Eastern Virginia. We hypothesize that the rural study location will have increased diversity and abundance, likely because of the continuous habitat availability. We predict urban environments to have lower values because of limited habitat availability and size. We analyzed camera trap data and identified species and behaviors that were placed in camera traps at Crow’s Nest Natural Area Preserve and the Virginia Outdoor Center during the fall of 2023 in Eastern, VA. Preliminary observations show that species abundance is higher in rural settings compared to the urban study location at the Virginia Outdoor Center. This research contributes valuable understandings of species interaction and presence in an urban and rural ecosystem to develop effective management plans for biodiversity conservation. As urbanization spreads into rural areas, understanding species abundance and richness dynamics is essential for developing strategies to protect and preserve vital habitats and ecosystems.
  • Poster 16: Effect of Food Source Nutritional Content on Little Black Ant Foraging Behavior
    Kylie Steffenhagen

    Interactions between species and their role in food webs are integral to ecosystem dynamics, and yet are areas of ongoing research in which our understanding is limited. The foraging behavior of ants provides an opportunity for insight into the function of food webs at the smallest scale. How does the nutritional content of different available food sources influence the foraging behavior of the Little Black Ant (Monomorium minimum)? This study was conducted in both forest and open field sites of the University of Mary Washington campus, near the Jepson Science Center. Three food samples of varying nutritional content – including tuna (protein), jelly (sugar), and cookies (protein & sugar) – were set out and observed. Data recorded from the Fall 2023 study, as well previous years (2019-2022), was compiled and analyzed for ant abundance patterns. It is expected that that the Little Black Ant, which is known to be an omnivorous scavenger, will display a preference for food samples containing both protein and sugar. By achieving a greater understanding of the foraging habits of species like the Little Black Ant, we can identify the role of each species in its larger food web, and we develop the tools required to undertake effective ecosystem conservation and restoration efforts.
  • Poster 20: Does the amount of rain effect the number of fish caught each year in alum spring?
    Rebecca Baker

    Rainwater that falls onto the ground collects particles and nutrients (nitrogen and phosphorus (Gilbert et al., 2005)) as it flows across the ground and into either aquafer underground or storm drains to streams. These nutrients then make their way into waterways where the nutrients can accumulate and produce algae blooms. Algae is good at a moderate amount because they are food for fish; either directly in the case of central stonerollers (Sublette et al., 1990) or indirectly being food for insect/insect larvae which are food for fish (; Pinder, 2023). Using annual rainfall data compared to the number of fish that were caught at alum spring creek every year, it is hypothesized that the more rainfall that there has been in a season, the more fish that will be caught. I expect that for years with a higher amount of rainfall for there to be more central stonerollers, bluegill, and fallfish caught in Alum Spring. Uncontrolled increase in nutrients in waterways can lead to an overproduction of algae. This overproduction of algae will cause eutrophication, which leads to fish die-offs from lack of dissolved oxygen and can have catastrophic effects on the marine ecosystem.
  • Poster 24: Temporal changes in day-night distribution of animal taxa
    Paul James Solis

    Many mammals are either diurnal or nocturnal, although these are not restrictive conditions. Species who may have an innate preference for certain hours may shift over time due to competition and/or predation. The ratio of species found in daylight to total species in both day and night was then calculated to estimate this preference. Individual trail cams were placed separately within Virginia Outdoor Center during Fall 2023. Footage of nearby peanuts and surrounding open area was taken at time intervals. Species data (either day or night) from 2017, 2019, 2021 was used for Crow’s Nest; data from 2019, 2020, 2022, 2023 was used for Virginia Outdoor Center. The null hypothesis was that there were no changes in day-to-total ratio of all animal taxa, even after some species were introduced in later years. The results showed that for all animal taxa at VOC, the slopes of the regression lines were significantly close to 0. As for Crow’s Nest, squirrels and coyotes had significant p-values (0.035 and 0.004, respectively), while the other taxa had insignificant findings. Linear regression analysis was able to determine significance via p ≤ 0.05. Determining potential differences between individual species under a taxa may be needed, but most taxa as a whole did not change in day-to-total ratios. 
  • Poster 27: Predicting Fish Populations: Do Longer Fish one year Predict More Fish the Next year?
    Samuel Gibbins Brown, Samuel Preston Trevey

    Modeling populations and predicting population changes year to year are important questions that biological scientists and ecologists attempt to answer across a variety of plant and animal populations of concern. Researchers will analyze available data from multiple yearly surveys of Hazel Run creek in Fredericksburg, Virginia and attempt to correlate mean fish length of a given species with the number of fish of the same species surveyed in the next year. Previous research from Barneche et al indicate that larger fish spend disproportionately more energy on reproduction than do smaller fish. Hazel Run survey data do not include weight, so researchers propose using length, a metric that was collected in the survey data, in an attempt to test this assertion. Evidence supporting the hypothesis will be positive correlations in data comparing mean fish length one year to number of fish of the same species surveyed the next year. Researchers predict that longer fish will on average weigh more, and if Barneche et al are correct, these fish will produce more offspring on a year to year basis.
  • Poster 32: Effect of Different Fishing Methods on Various Species of Fish over Time
    Dewey Snyder

    Over the previous seven years, both seining and electrofishing have been used to fish in Hazel Run, both with varying success rates on the species that inhabit Hazel Run. Time is not a considered factor in the equation, and so the calculations run tests whether or not the percent of certain fish species caught via seining vs. electrofishing has significant variance over the course of seven years. To test this, five species of fish present across all seven years were chosen and the catch numbers were converted to percentages for each type of fishing. The effectiveness percentages were then compared on the scale of time from 2016 to 2023. Of the five species examined, the bluegill had percentages that remained relatively steady across seven years; the fallfish and the largemouth bass had percentages that fluctuated across seven years with no discernable pattern; and the bluehead chub and the redbreast sunfish had potentially significant rises and falls in the percentages across seven years.
  • Poster 35: Mammal’s Recorded on Trail Cameras in Relation to Distance from Human Habitats
    Hunter Garnett

    The encroachment of human development onto forested areas will increase the interactions between humans and animals. Previously published research by (Soergel) states that human disturbance can alter the behavior of mammals to become bolder in the face of human development as in the case of raccoons, or they can avoid dense human development as in the case of bobcats. I aim to determine if distance from human development has an effect on the mammalian species captured on trail cameras. To test my hypothesis that cautious mammals prefer to distance themselves from human development, trail cameras were set up on the Virginia Outdoor Center’s wooded trail. Each trail camera was placed several meters off of the trail and into the woods positioned one to two feet off of the ground. In order to increase the success of capturing mammals on the trail cameras whole peanuts were used as bait. The trail cameras were then left out for exactly one week to collect images. I expect that the data collected from the trail cameras will support my hypothesis that there would be a difference in mammalian species captured based upon the distance from human development.   
  • Poster 45: The correlation between fish length and pass number
    Aleksandra Witkowski

    Fish can vary in size and withstand different amounts of electricity depending on length. Larger fish are known to withstand more voltages of electricity when compared to smaller species. This topic addresses the benefits of using electrofishing as a sampling method and helps to identify if the average length of species caught is affected while using this method. This study asks the question of if there is a correlation between the length of fish caught and the pass number in which they were captured. Data was collected using an electro fisher, nets, buckets, as well as a solution that temporarily calmed the fish in order to be measured. A sample area was measured out in the Hazel Run Creek, and a pass was done through the water in the sample area in order to collect data. The data shows that there is no noticeable trend in which the pass number relates to the average length of species caught. The hypothesis was not supported, and as pass numbers increase, there is no correlation to the average length of species caught. Based on the data, when using the electrofishing technique there is no disadvantage as a sampling method when collecting average lengths. 

BIOL 432 Virology, Dr. Lynn Lewis
In this Research Intensive course, students are introduced to the major groups of viruses and their replication cycles, lab techniques for studying viruses and some of the new, emerging viruses.  Students practice lab techniques for handling, growing and studying viruses during the first half of the semester, while designing an experimental research project based on their reading of primary literature.  Students then carry about their research projects, in groups of 2 or 3, during the second half of the semester and present their collected data analysis.

The following are abstracts for the projects presented in this symposium:

  • Poster 3: Antiviral Effects of Chlorpromazine Against NDV in BHK-21 Cells
    Aisling Berrios, Jenny Vuong, Cyrus Ramsey

    In today’s world, with the COVID-19 pandemic and extremely high antiviral prices, there has been a push for new, novel antiviral treatments. Newcastle Disease Virus, or NDV, is a detrimental virus to poultry and can cause extreme economic loss for the poultry industry. Our research project aimed to investigate the properties of the antipsychotic drug, chlorpromazine, as an antiviral against NDV in BHK-21 hamster cells. The goals of our research were two-fold, to contribute to the current knowledge of NDV treatments, and to potentially relate this to other viral respiratory diseases such as COVID-19. We hypothesized that chlorpromazine will exhibit antiviral effects against NDV, resulting in an decrease of cell death due to NDV infection. To do this, we performed a TCID50 assay to determine the cytotoxicity of the drug using chlorpromazine concentrations of 5, 7, and 9µM. Due to difficulties with the incubator, we were delayed in our experimental procedures. Thus, we will be performing an MTT assay at a later time to investigate the potential benefits of using chlorpromazine as a treatment. Statistical analysis of this test will be done through a one-way ANOVA. Our cytotoxicity assay found that none of concentrations were cytotoxic to the cells, with chlorpromazine at a 5µM concentration proving to be the most viable. Analysis is ongoing.
  • Poster 6: Antiviral Effects of Echinacea purpurea on NDV in BHK-21 Cells
    Jessie Mills and Kinsey Brotman

    Newcastle disease virus (NDV) is a deadly paramyxovirus that can affect the respiratory system in chickens. E. purpurea is an herbal supplement that has been proven through numerous studies to have antiviral properties. Expanding research on the antiviral effects of E. purpurea is
    important because it is easily accessible and may help decrease viral spread. The question being tested is if E. purpurea can act as an antiviral against NDV in BHK-21 cells. An initial cytotoxicity assay was performed using different dilutions of E. purpurea extract to ensure that the extract chosen would not have cytopathic results on BHK-21 cells. The results of the assay showed that the extract is safe for the cells as it did not have any cytopathic results. An MTT assay will then be completed in order to determine if a 10-3 dilution of E. purpurea extract has an effect on BHK-21 cell viability when treated with various NDV dilutions. If cells treated with the extract have less cell death post viral infection compared to the cells with no extract treatment, our prediction that E. purpurea has antiviral effects on NDV can be supported. The treatment of NDV with  E. purpurea can help show how the natural extract works as an antiviral. 
  • Poster 18: The Antiviral Effects of Elderberry Extract on Newcastle Disease Virus (NDV) 
    Caeley Terapane, Tyler Michael, and Melissa Garcia

    This study will focus on determining the antiviral activity of elderberries on the Newcastle disease virus. BHK-21 cells will be infected with Newcastle disease virus (NDV) and treated with different concentrations of elderberry extract in order to determine the antiviral effects of elderberries on NDV-infected cells. If elderberry extract can be used to treat influenza and the common cold, other viral infections such as NDV should have the same effect when introduced to elderberry. Elderberry contains compounds, such as flavonoids and anthocyanins, which are believed to have antiviral proteins.  Some studies also suggest that elderberry may stimulate the immune system, which could help the body defend itself against viral infections. An MTT cytotoxicity assay will be performed with the main goals of determining what effects elderberry extract has directly on the proliferation and viability of BHK-21 cells, and determining what concentration of extract will be safe to use on these cells. Another preliminary TCID50 cytotoxicity assay will be performed to determine the concentration of NDV virus suspension required to infect half of the BHK-21 cells in the cell culture. Once the initial MTT assay and TCID50 assay have been performed, a plaque reduction assay will be conducted with the main goal of determining the antiviral effects of elderberry extract on the late-stage infection of Newcastle Disease Virus (NDV) in BHK-21 cells. We expect the results from these tests to indicate that elderberry extract when introduced to NDV-infected BHK-21 cells, will have direct antiviral properties against the spread of infection and promote the proliferation of the cells themselves.  
  • Poster 21: The effect of Matricia chamomilla on Newcastle Disease Virus on BHK-21 cells
    Jaylyn Long, Tyler Lowe, Rose McMullen

    Newcastle Disease Virus (NDV) is a virus that attacks poultry’s digestive, nervous, and respiratory systems. There has been substantial research investigating vaccination and antibiotics of this virus; however, these techniques have been proven to be less useful over time. Our research project seeks to test if chamomile extract has an effect on NDV proliferation in BHK-21 cell cultures. We hypothesize that by introducing the chamomile extract, German chamomile (Matricaria chamomilla) to BHK-21 cells infected with NDV, the number of cells killed by the virus will decrease. To test the cytotoxicity of the chamomile extract on BHK-21 cells, an MTT assay will be performed. Six different dilutions of the chamomile extract, ranging from 10^-2 to 10^-8, will be tested and compared to the control: BHK-21 cells without chamomile. A second MTT assay will be performed to analyze the effectiveness of the chamomile treatment on virus proliferation. Tolerable concentrations of chamomile will be tested and compared to the control, BHK-21 and NDV, and BHK-21 cells without the virus or treatment. The viability of the treated and untreated cells will then be evaluated and compared. The cytotoxicity test revealed that all dilutions of chamomile extract are tolerated by the BHK-21 cells. We plan on performing an MTT assay using the 10^-2  dilution of chamomile and different concentrations of NDV. Using a one-way ANOVA test, we expect that the treated cells will have a higher absorbance than the untreated cells, indicating that chamomile-treated cells have a higher metabolic rate than untreated cells when infected with NDV. 
  • Poster 25: Using Different Concentrations of Ginger Root Extract derivative to Test the Effect on Encephalomyocarditis Virus in BHK-21 cells as a Pre-treatment, Co-treatment, and Post-treatment
    Lordina Agyekum, Arianna Chase, Marion Robinson 

    Encephalomyocarditis virus is a known livestock virus that particularly affects pig farms as well as other mammals through contaminated food and water. It is a known zoonotic disease that can infect humans and cause physiological symptoms if infected. Ginger Root has gone under recent investigation in antiviral therapy with showing abilities to attenuate viral replication pathways. Its derivative, Gingerenone A, is highly prevalent in the root and will be used in this experiment as we plan to observe it as a form of treatment for Encephalomyocarditis virus (EMCV) in baby hamster kidney (BHK-21) cells. We will test if Gingerenone A has any effect on EMCV infectivity when used during pretreatment, co-treatment, and post-treatment of the virus. We hypothesize that if given a range of different concentrations of ginger extract, the highest concentration (20 micromolar) of the Gingerenone A will decrease infectivity of EMCV in the three treatment groups. Through the measure of cell viability by MTT assay and virus infectivity of TCID50 assay, we will be able to analyze and determine the constraints that decrease cell death and increase cell viability compared to the control group of EMCV infected BHK-21 cells.
  • Poster 29: The Effect of the Bloodroot Alkaloid, Sanguinarine, on Newcastle Disease Virus
    Ciara Donnangelo and Delaney Baratka

    Sanguinarine, a natural alkaloid of bloodroot, has been shown to have antiviral potential in past studies. For this experiment, we want to test these properties using the Newcastle disease virus (NDV) to infect sanguinarine-treated BHK-21 cells. Both sanguinarine and NDV have an effect on the molecular target NF-kB signaling pathway, which controls cell viability. This leads us to believe that sanguinarine may be a probable treatment for NDV. To test this, we plan to perform an MTT assay to determine cell metabolic activity and cytotoxicity. We also plan on a cell viability assay to measure cell survival. The main purpose of this study is to gain a better understanding of the antiviral and cytotoxic properties of sanguinarine. 

BIOL 445 Research Practices in Aquatic Ecology, Dr. Abbie Tomba
Students conduct original research projects focusing the ecology of local stream organisms such as aquatic invertebrates and algae.  Student projects can be lab or field based  and often answer questions addressing how different  factors affect individuals,  populations, or the biodiversity of these different groups.

The following are the projects presented in this symposium:

  • Poster 2: Flow rate effect on Parasitism in the Rappahannock River
    Taylor Nicholson, Jenny Vuong, Kaitlin Butler, Joshua Rowe

    Aquatic freshwater snails are grazers that are intermediate hosts for many parasite species like trematodes, which utilize the host’s body to their advantage, changing the host’s distribution patterns to increase parasitic transmission. This relationship is important for ecosystem dynamics and understanding where this relationship occurs most frequently. We hypothesized that low-flow areas of the Rappahannock River in Fredericksburg, Virginia have a higher prevalence of parasitism due to previous findings that there’s a higher overall snail density in pools. We chose a total of 16 1x1m areas in pool and riffle habitats of the river to collect the snails and test abiotic factors, including shade, water temperature, pH, %DO, and flow rate. We then dissected the snails, identified as Elima and Leptoxis, to determine parasite status and species. So far, we have found that neither shade (T-test, t = 1.051, df = 14, p = 0.844) nor water temperature (Two-sample Wilcoxam = 16, p = 0.052) differed significantly between the two habitat types. Of the 240 snails collected, there was an average of 0.00 ± 0.00 (n = 90) parasites found in the snails of the riffles whereas the pool snails had an average of 3.50 ± 1.09 (n = 150) parasites. Further analysis using T-tests to determine differences between pools and riffles regarding other abiotic factors and parasite and snail species will be conducted. We will also complete a linear regression to indicate the relationship between infected snail abundance and flow rate. At this time, analysis is ongoing.
  • Poster 10: Trematode parasite prevalence in Elimia virginica snails based on water depth
    Eliana Ramirez, Rosie Stewart, Noah Smith, Anias Saunders

    Freshwater snails live in shallow or deep areas of water where they can be infected by trematode parasites. Trematodes are able to live anywhere and are known for transmitting parasites to snails to be their temporary host. Parasites have the ability to manipulate a snail’s behavior and movement by moving snails closer to the water’s surface risking predation and infection. We decided to conduct an observational study to investigate the effects of trematode parasites on Elimia virginica snails in the Rappahannock River, specifically comparing the prevalence of parasites in shallow vs. deep water. We sampled 10 shallow and 10 deep pools where 10-100 snails were collected for each area. Pools were one by one meters in size and a depth of below or above 0.3 meters. Other factors were also measured for consistency such as pH, DO, temperature, and shade. Once snails were collected, they were placed into an ethanol bag and refrigerated in a lab room. During lab processing, snails were measured and recorded in width and then dissected to look for parasites. Results demonstrated that there were six infected snails in shallow pools and only three in deep, supporting our hypothesis of snails residing in shallow pools. Parasitized snails in shallow were 7.15% compared to 7.49% in deep pools. Snails that were larger in width had more parasites than smaller ones. This study could be useful for future references in behavioral patterns of E. virginica snails in freshwater habitats when infected by trematodes.
  • Poster 15: The Relationship Between Runoff Nitrogen and Periphyton Biodiversity in the Rappahannock River.
    Alan Fletcher, Caitlin Moore, Ellie Shanahan

    Researchers across the globe have sought to study the effects of large-scale agriculture on aquatic environments. This study sought to observe the relationship between varying levels of fertilizer-based nitrogen and the biodiversity of periphyton in the Rappahannock river. Periphyton refers to the cultures of microscopic producers found on a variety of aquatic surfaces. As primary producers, periphyton form a foundation on which a freshwater ecosystem rests. The researchers hypothesized that as nitrogen levels increased, so would the biodiversity of observed periphyton. This hypothesis was supported by the rationale that nitrogen, as a crucial element for autotrophic development, would have a measurable impact on periphyton proliferation if altered. To answer the question posed by the researchers, a singular monoculture of periphyton was collected from the Rappahannock river and divided into 25 samples. These sample cultures were separated into 5 groups. These groups would be exposed to one of five treatment levels. The treatment levels were 0x, 0.5x, 1.0x, 1.5x, and 2.0x the baseline nitrogen level of the Rappahannock river. After 2 weeks of exposure, the 25 periphyton samples were observed under a microscope and their taxa and number of individuals within them recorded. The researchers found that the periphyton cultures exposed to the baseline nitrogen levels displayed the greatest variety of taxa and number of individuals when compared to the other treatments. This did not support the initial hypothesis made prior to the experiment.

BIOL 472K Research in RNA Technology, Dr. Ginny Morriss
This course exposes students to the world of gene expression through the lens of RNA. We cover the biosynthesis, processing, and function of different RNA types and explore how RNA biology can be leveraged for use in research applications. Students learn basic techniques that are used in RNA laboratories throughout the world and design projects that utilize RNA technologies to study a question of interest.

The following are abstracts for the projects presented in this symposium:

  • Poster 5: The Relationship Between CRISPR-Cas9 Dop2R Gene Knockout and Climbing Velocity in Drosophila melanogaster
    Jessie Mills, Analisa Ayala, Yamila Merida

    Dopamine receptors play a role in human learning abilities, hormonal control, and locomotor function. Knockout of the human DRD2 gene is associated with diseases such as Parkinson’s, which causes mobility issues. The ortholog Dop2R plays a similar role in Drosophila melanogaster, working as a dopamine inhibitor to regulate the mobility of the fly. In our experiment, we are examining the effect of Dop2R knockout on Drosophila climbing ability. By targeting the Dop2R gene, dopamine suppression is no longer possible, and excitable neurons will continuously fire, stimulating action potentials and causing the impairment of fly climbing ability through tremors, stiffness, or in extreme cases paralysis. We completed crosses between virgin female UAS-Cas9.P2/CyO; tub-Gal5/TM6B1Tb1 and male Dop2R sgKO/Dop2R sgKO. From the F1 generation, we were able to collect UAS-Cas9.P2/+; tub-Gal4/Dop2R sgKO experimental flies and control flies. We measured climbing velocity (mm/sec) in the Dop2R KO flies compared with control groups separated by sex. ImageJ will be used to analyze climbing velocity data using a two-way ANOVA. To confirm knockout efficiency, RNA will be isolated, followed by RT-PCR and gel electrophoresis. We expect to see lower average climbing velocity for the Dop2R KO groups compared to the controls. If the Dop2R KOs have lower average climbing velocities, we can support our prediction that the knockout of Dop2R impacts climbing abilities. The knockout of the Dop2R gene can help show how motor function is affected in mobility disorders such as Parkinson’s disease.
  • Poster 11: Analyzing the Relationship Between Oxidative Stress Levels and SOD1 Using Drosophila Melanogaster Relating to ALS
    Evelyn Buehren, Charlotte Russell

    The primary objective of this research study is to analyze the relationship between oxidative stress levels and SOD1 in relation to ALS. Amyotrophic Lateral Sclerosis (ALS) is a rare neurological disease that causes motor neuron death, resulting in muscle atrophy. There is no known cure for ALS and only two therapies have been approved by the FDA. Mutations in the SOD1 gene, a regulator of oxidative stress, have been linked to ALS. We generated neuron-specific, temperature sensitive SOD1 RNAi knockdown and CRISPR/Cas9 overexpression flies to study muscle function and oxidative stress compared to wild type flies. To avoid embryonic lethality, flies were incubated through the embryonic and larval period at 25 degrees C and switched to 30 degrees C during the pupal stages for one week to activate the knockdown and overexpression. We measured climbing velocity (mm/sec) as a proxy for motor function. We will measure oxidative stress levels using a marker, the mpk2 gene. RT-qPCR will be conducted to analyze the expression of SOD1 and mpk2 in the knockdown, overexpression, and control groups. The experimental groups will be compared to a control group of wild-type Oregon R. We expect to find decreased motor function in both overexpression and knockdown groups compared to the control group because altering expression of the SOD1 gene is toxic to motor neurons. We expect the RT-qPCR to show increased oxidative stress in the experimental groups, compared to the control, indicating that SOD1 mutations are linked to ALS due to oxidative stress mechanisms.
  • Poster 17: MBNL1 Downregulation Affects Celf1 mRNA Decay by Alternative Splicing and Polyadenylation
    Samantha Schwarting, Arshpreet Brar, Orianne Mbuyi Mujinga Kazadi,

    Myotonic dystrophy type 1 (DM1) is a disease characterized by skeletal muscle weakness and myotonia. DM1 is caused by the expansion of CTG repeats in the 3’ untranslated region (UTR) of Dystrophia myotonica protein kinase (DMPK) gene. In DM1 patients, Muscleblind like 1 (MBNL1), an RNA binding protein, is sequestered at the CTG repeats while CUGBP Elav- Like Family Member 1 (CELF1), another RNA binding protein, is upregulated. MBNL1 affects alternative splicing and alternative polyadenylation in the 3’ UTR of CELF1. Given the role of MBNL1 in mRNA decay and upregulation of CELF1 proteins DM1, it is expected that MBNL1 facilitates the decay of Celf1 mRNA by promoting the formation of less stable Celf1 mRNA isoforms. We used siRNAs to knockdown Mbnl1 and we will look at Celf1 decay over time in C2C12 cells. We harvested cells at 0, 2, 4, and 6 hours after actinomycin D treatment, a drug that inhibits mRNA synthesis. After harvesting the cells, we isolated RNA, performed RT-PCR, and we will perform polyacrylamide gel electrophoresis to look at the amount of different Celf1 mRNA isoforms and the total Celf1 mRNA present at each time point. We will use ImageJ to analyze gel electrophoresis results and confirm MBNL1 knockdown using RT- qPCR. Upon the successful knockdown of MBNL1, the experimental group will show a slower decay rate of Celf1 mRNA compared to the controls. The results will help us determine if MBNL1 regulates CELF1 proteins through mRNA decay.
  • Poster 33: The Effect of Apc/Apc2 KO on Drosophila melanogaster Colorectal Tumor Growth
    Willa Clore, Alexis Miller

    The adenomatous polyposis coli (APC) gene is a tumor suppressor gene composed of the Apc-like and Apc2 genes that regulates cell growth and prevents proliferation of metastatic colorectal cancer cells. Drosophila Melanogaster is a model organism for analyzing the effects of functional changes of tumor suppressor genes because of the shared genomic and morphological similarities with the mammalian signaling pathway. To study the effects of decreased APC function, the experimental flies will undergo CRISPR/Cas9 knockout (KO), and those with the APC gene intact will be used as the control. Flies with ubiquitous Apc/Apc2 knockout, driven by alpha-tubulin 84B regulatory sequences resulted in lethality of all experimental flies. These results indicate that the Apc/Apc2 CRISPR/Cas9 KO worked effectively, as hypothesized. To address this lack of an experimental group, Apc/Apc2 sgRNA KO flies will be driven using skeletal muscle-specific myosin heavy chain regulatory sequences, and the offspring will be used as the experimental group.  Both the experimental and the control groups will contain subgroups that will either consume 2mM acrylamide or 7.5% EtOH for ~3 days, or not have any carcinogen exposure. After exposure, tumor growth will be quantified using microscopy, followed by RNA isolation.  The RNA will then be analyzed to study the effect the APC gene has on tumorigenesis (i.e. initation or progression) of the Drosophila m. colorectal tract.  With this data, a 2-way ANOVA will be performed to determine the role of the APC gene on colorectal tumor growth in control groups compared to carcinogen-exposed groups.
  • Poster 36: The Effect of GlyS Over-Expression on Drosphila melanogaster Exercise Tolerance
    Owen Reilly, Nathan Terrell, Peyton Garst

    Generalizing alterations in skeletal musculature solely due to hemodynamic deficiencies and undermining enzymatic activity, including glycogen synthase, has led to inefficient treatment of exercise intolerance. Glycogen synthase is crucial in the anabolic pathway of glycogen deposition and is required for the maintenance of muscle’s metabolic fuel. We developed an inducible, muscle-specific Drosophila melanogaster (fruit fly) model over-expressing glycogen synthase (Glys), resembling its human ortholog, GYS1. We hypothesized when subjected to overexpression, Drosophila melanogaster will present observable increased exercise tolerance. Virgin female overexpressed Glys flies were crossed with male UAS-Cas9.VPR,tub-Gal80/Mef2R-Gal4/Tm6Bb,Tb flies. Experimental flies were isolated and are currently being incubated for one week at 30 degrees Celsius. Next, a negative geotaxis assay will be performed, testing the fly’s ability to perform adequate locomotion up seven cm during a five-second duration. Successively repeating the assay will allow for the analysis of flies’ exercise tolerance by comparing the number of flies that reach the designated mark each trial. If a steady increase in pass rate is documented then it may be inferred that overexpression of Glys is effective in increasing exercise tolerance. To confirm the data collected is a direct result of Glys overexpression RNA isolation and RT-qPCR will be conducted; thus, ensuring anticipated genomes were properly manipulated. Upon completion of this study, a more developed understanding of exercise intolerance due to alteration glycogen synthesis may be established; potentially aiding current endeavors to formulate effective treatments.