Understanding the effects chemical exposure has on neurodevelopment is crucial for protecting human health and improving environmental policies and safety. Our research uses zebrafish to investigate how environmental and chemical exposures influence early development, as their embryos allow us to observe changes across multiple biological levels. Within our collaborative lab structure, research teams work together to examine how various factors affect development from genes to behavior. My role in the lab includes training in zebrafish-based experimental techniques and beginning to conduct literature analysis to identify candidate chemicals for future experiments.
Plants have evolved a sophisticated set of pathways to detect and respond to light, which allows them to adjust their development in response to changing conditions. Red and far-red wavelengths are detected by photoreceptors called phytochromes (phys), with phyB being the major phytochrome involved in red-light response in the model plant Arabidopsis thaliana. Phytochrome levels are regulated by an E3 ubiquitin-ligase complex that includes the target-adaptor Light-Response BTB1 or BTB2 (LRB1 or LRB2) proteins. The Gingerich lab studies lines of Arabidopsis that contain mutations in the LRB1, LRB2, and PHYB genes. Analysis of growth responses to light and other environmental or physiological factors that intersect with light response helps us better understand how the phytochrome pathway regulates development. Here, we present an analysis of germination responses to red and far-red light and seedling development responses to the hormone methyl jasmonate in our lines. Germination response to red and far-red light is well-studied, and recent analyses have suggested roles for phyB in modulating jasmonate responses; thus, studies of these responses in lines with alterations of the phytochrome pathway might be informative.
Rare diseases affect 30 million Americans, many of whom remain undiagnosed due to limited functional characterization of DNA variants. Propionic acidemia is caused by variants in PCCA or PCCB that impair enzyme function, leading to severe metabolic dysfunction, often presenting in early infancy. While the Wisconsin newborn screening panel tests for this disorder, screening is neither 100% effective nor does it identify the cause of propionic acidemia in each patient. There are 979 reported DNA variants of uncertain significance or conflicting classification in PCCA and PCCB, meaning that it is unclear if these mutations cause the disease: thus, identification of one of these variants in a patient does not equal clear diagnosis. To address this gap, our lab uses a minigene system to examine whether variants have functional effects. Although we can effectively assess individual variants with this system, it is a relatively low-throughput method. We present our efforts at optimizing this system through improved sample processing, next-generation sequencing (NGS), and development of efficient R scripts. An improved pipeline should accelerate the resolution of variants of uncertain significance associated both with propionic acidemia and across rare genetic diseases.
Schistosomiasis is a neglected tropical disease that affects over 250 million people worldwide and is caused by parasitic flatworms known as schistosomes. Miracidia, the first larval stage of schistosomes, infect snails as intermediate hosts, where they mature into a larval stage capable of infecting humans. Although it is not definitively known how miracidia locate snails, they have been shown to detect and interpret light to navigate their environment. The purpose of this study was to analyze the movement of Schistosoma mansoni miracidia in response to the presence of light and to different wavelengths of light. Miracidia were loaded onto rectangular arenas and exposed to light gradients, including white, red, blue, and/or green light. The miracidia were recorded for 1 hour and tracked using custom code. As expected, and consistent with previous work, miracidia are photoattracted. Notably, preliminary results indicate that they prefer blue over red light but have no apparent preference between blue and green light. Future experiments will explore the integration of their light and chemical perception. Understanding this sensory coordination could be key to developing new strategies to reduce schistosome populations and the spread of schistosomiasis.
Schistosomiasis is a neglected tropical disease caused by parasitic flatworms of the genus Schistosoma, affecting over 250 million people worldwide each year. These parasites use freshwater snails, such as Biomphalaria glabrata, as intermediate hosts to complete their life cycle. There are potential gene candidates of interest that may play a role in the chemosensory attraction of the worms, allowing them to seek out the snail. Current methods for genome editing in B. glabrata are lacking. This research investigates the methodical development of an ex ovo culturing method for B. glabrata embryos and juveniles. Establishing this system is a key step toward future genome editing studies targeting candidate genes in the snail, enabling assessment of how disruption of these proteins may affect worm attraction. If transgenic snails with targeted gene knockdowns that prevent parasite-host recognition were introduced, a gene drive could spread these traits through wild populations, potentially reducing transmission and lowering the burden of schistosomiasis.
Schistosomiasis is a neglected tropical disease caused by three parasitic flatworm species, including Schistosoma mansoni, that require a freshwater snail intermediate host. Disrupting parasite host seeking behavior by miracidia, the first larval stage responsible for locating and infecting a snail host, could provide a novel disease control strategy. Praziquantel, the current treatment of choice, targets a transient receptor potential (TRP) channel, though its mechanism is not linked to sensory disruption. However, because TRP channels function broadly as metazoan sensory receptors, we hypothesized that modulating these channels could impair miracidia host seeking abilities. We developed a high throughput phenotypic screening platform using a custom acrylic 96-well screening arena and a high resolution, multicamera wide field imaging system to quantify miracidia behavior. In a primary screen of 128 TRP targeting compounds, 14 hits were identified based on altered motility or behavior. These hits were subsequently evaluated in dose response assays across multiple concentrations. Future studies will characterize the molecular mechanisms of active compounds and evaluate their effects on host seeking and infection. Targeting TRP mediated sensory behaviors may provide a novel strategy to disrupt the schistosome life cycle and reduce infection.
Schistosomiasis is a neglected tropical disease that affects over 250 million people worldwide and is caused by parasitic flatworms known as schistosomes. Miracidia, the first larval stage of schistosomes, infect snails as intermediate hosts, where they mature into a larval stage capable of infecting humans. Although it is not definitively known how miracidia locate snails, they have been shown to detect and interpret light to navigate their environment. The purpose of this study was to analyze the movement of Schistosoma mansoni miracidia in response to the presence of light and to different wavelengths of light. Miracidia were loaded onto rectangular arenas and exposed to light gradients, including white, red, blue, and/or green light. The miracidia were recorded for 1 hour and tracked using custom code. As expected, and consistent with previous work, miracidia are photoattracted. Notably, preliminary results indicate that they prefer blue over red light but have no apparent preference between blue and green light. Future experiments will explore the integration of their light and chemical perception. Understanding this sensory coordination could be key to developing new strategies to reduce schistosome populations and the spread of schistosomiasis.
Light is an important resource for plants, as it serves both as energy and as a signal for growth and development. For this reason, plants have evolved a complex system of signaling pathways to sense and react to light in many conditions. Red and far-red light are detected by photoreceptors called phytochromes. The phytochrome phyB has been the target of mutant studies by Gingerich lab. One of the phyB mutant lines isolated by the lab displays an increased response to both red light and blue light. In this line phyB is predicted to contain a change in a single amino acid as a result of a single nucleotide polymorphism (SNP) in the PHYB gene. To confirm that it is indeed the phyB mutation in this line that is causing the red and blue light phenotypes, we are implementing a CRISPR-Cas9-based gene editing system for Arabidopsis in the lab, with the purpose of recapitulating the PHYB SNP of the mutant line in plants that are otherwise wild-type. The specific system we are using is “prime-editing”. We will detail synthesis of the DNA constructs needed to implement the prime editing and our current efforts to transform plants and initiate the editing process.
Plants have evolved a sophisticated set of pathways to detect and respond to light, which allows them to adjust their development in response to changing conditions. Red and far-red wavelengths are detected by photoreceptors called phytochromes (phys), with phyB being the major phytochrome involved in red-light response in the model plant Arabidopsis thaliana. Phytochrome levels are regulated by an E3 ubiquitin-ligase complex that includes the target-adaptor Light-Response BTB1 or BTB2 (LRB1 or LRB2) proteins. The Gingerich lab studies lines of Arabidopsis that contain mutations in the LRB1, LRB2, and PHYB genes. Analysis of growth responses to light and other environmental or physiological factors that intersect with light response helps us better understand how the phytochrome pathway regulates development. Here, we present an analysis of germination responses to red and far-red light and seedling development responses to the hormone methyl jasmonate in our lines. Germination response to red and far-red light is well-studied, and recent analyses have suggested roles for phyB in modulating jasmonate responses; thus, studies of these responses in lines with alterations of the phytochrome pathway might be informative.
Unionidae, also known as freshwater mussels are a surprisingly charismatic species, every step of their life history is strange and intriguing. While odd, freshwater mussels are vital ecosystem engineers and play significant roles in nutrient cycling within their aquatic environments. Due to changing freshwater habitats they are considered to be the most endangered family of animals in North America. Unfortunately, humans have often overlooked them, either when walking through a stream or through gaps in research knowledge. In order to fill in a part of that hole, we conducted qualitative surveys in distinct streams throughout Chippewa county Wisconsin to determine the distribution and composition of the mussel population within the county.We found 17 total species of the 50 previously known species native to the area. These results are the first step towards a holistic picture of the current patterns of mussel distributions within the Chippewa River Watershed.
Understanding how closely related species respond to ecological variation and natural selection is central to predicting evolutionary change, especially under rapidly shifting environmental conditions. This study uses perennial sunflowers (genus Helianthus) in the Upper Midwest as a focal system for examining trait differentiation and selection dynamics across time and species. We established multi-year common gardens at two sites in Eau Claire, WI, to experimentally compare functional traits and fitness responses among three widespread Helianthus species: H. giganteus, H. grosseserratus, and H. maximiliani. These species co-occur across the region and often overlap in distribution, where trait divergence may mediate niche partitioning or reflect variation in selective pressures. Comparing selection patterns across species, sites, and years can help evaluate how evolutionary history, resource availability, and climate variation shape trait evolution in wild sunflower populations. This research contributes to broader ecological and evolutionary questions about phenotypic plasticity, adaptation, and species coexistence, providing a long-term dataset for understanding plant responses to a changing environment.
Fruit agriculture relies on insects to provide the regulating ecosystem service of pollination. For fruit to successfully set, a pollinator must contact the stigma to deposit pollen. Sweet cherry (Prunus avium) is a pollination-dependent fruit cultivated globally and is a growing industry in Australia. Despite this, little work has been done to understand its pollinators which include native bees, hoverflies, and the introduced western honey bee (Apis mellifera). By observing cherry flower pollinator visits, we can document the variety, duration, and frequency of on-flower behaviors, determine which taxa engage in stigmal contact, and investigate vulnerabilities of pollination service to loss of pollinator taxa. In this study, 68 video recordings (2,035 minutes of footage) of two sweet cherry cultivars (Lapins and Ron’s Seedling) in Bilpin and Young, New South Wales, Australia were annotated. Flower visitors were identified to the species level and included the orders Hymenoptera and Diptera. Results were analyzed to create taxon-specific foraging behavior radar charts, assess frequency of stigmal contact, and simulate extinctions. This work informs cherry cultivation and on-farm pollinator conservation.
Managed honeybees (Apis mellifera) are often employed by Wisconsin cranberry (Vaccinium macrocarpon) growers to improve crop yields, however honeybees are unable to perform buzz pollination. For cranberry pollen to be released, the anthers must be vibrated at a specific frequency. Bumble bees (Bombus spp.) can buzz pollinate and are more efficient pollinators of cranberry but are not as commonly utilized for commercial pollination. This study determined the frequency and duration of buzz pollination provided to cranberry flowers by visiting bumblebees in relation to the stigmal status of the cranberry flowers visited and the resource (pollen or nectar) being foraged for. Data was collected during the 2025 bloom period (June 11-July 10) using video and audio recordings to record the foraging behavior of flower visiting pollinators. The videos were then annotated to document flower age, pollinator identity, floral resource foraged for, and frequency and duration of buzz pollination. Bumblebee species B. bimaculatus and B.impatiens buzz pollinated more often than other species observed. Bumblebee species overall buzz pollinated roughly a third of a second longer when the stigma was exposed. By increasing our understanding of pollinator foraging behavior in cranberry, growers can make more informed decisions regarding pollinator management on their farms.
Urbanization can alter soil chemistry, potentially influencing floral nectar composition and pollinator foraging behavior. We examined how two micronutrients commonly associated with urban environments, sodium (Na⁺) and potassium (K⁺), affect pollinator foraging. We conducted 10-minute field observations of pollinator visitation to flowering plants in urban areas on flowers supplemented with nectar (sugar water (15%), with Na⁺ or with K⁺).Next, we performed laboratory foraging studies with butterflies (Vanessa cardui) and bumble bees (Bombus impatiens). Individuals were introduced to a foraging arena containing nectar sources with randomly assigned treatments (sugar water, with Na⁺, or with K⁺). The butterflies foraged from artificial flowers, and bumble bees from blue plastic chips each containing nectar, with treatment placement randomized each trial. Foraging activity was recorded using a video camera, and individuals were allowed to forage for 30 minutes. Afterward, recordings were annotated to quantify foraging activity.To date, we have conducted 15 field observations and 39 butterfly trials; the bumble bee trials are ongoing. We expect differences in visitation and nectar preference among treatments in both field and laboratory settings, including variation between butterflies and bumble bees. This work highlights both the ecological importance of nectar micronutrients and the importance of studying pollinator behavior.
Pollination is essential for farmers to produce high fruit yields in Wisconsin cranberry (Vaccinium macrocarpon) marshes, yet the foraging behavior of key pollinators such as bumblebees (Bombus spp.) and honeybees (Apis mellifera) remains poorly understood. While honeybees are commonly used as managed pollinators in cranberry marshes, limited research has compared their foraging behavior on cranberry flowers to native bumblebees. This study compares the foraging behavior of these two major pollinator groups to assess differences in behaviors like stigmal contact rates, visitation frequency, and changes in visit duration in response to temperature. Behavioral data was collected during the 2025 bloom period (June–July) using video recordings and human observations. Video cameras were set up throughout the marsh and footage was annotated to quantify stigmal contact and visit duration; temperature data was recorded when videos began and ended. Human observations of individual bees through the marsh were used to quantify the number of flowers visited per minute. The analysis showed that bumblebees contacted the stigma more often and had an increased visitation frequency compared to honeybees with temperature not having a significant effect between the species. Our findings inform pollinator management practices and enhance our understanding of species-specific contributions to cranberry pollination.
Azospirillum (AZO) and Gluconacetobacter (GLU) are two genera of naturally occurring bacteria that convert nitrogen, a vital plant nutrient, from its atmospheric form into a form usable by plants. AZO is associative and functions around the plant roots, whereas GLU is endophytic, working within the plant tissue. Nitrogen-fixing bacteria are gaining popularity in sustainable agriculture efforts as biofertilizers to provide an alternative to environmentally problematic synthetic fertilizers. Companies such as TerraMax Inc. are interested in finding the most effective bacteria for biofertilizers to optimize crop yield. This research compared the impact on corn growth and function of the more widely used AZO and the novel biofertilizer candidate GLU, both with varying application methods: soil drench, leaf coating, or a combination of those two. The results favor GLU over AZO for increasing nitrogen availability in corn and indicate the potential for GLU to improve yield through significantly increasing leaf net photosynthesis, chlorophyll content, and shoot biomass compared to the control when applied directly to the leaves. These results suggest Gluconacetobacter (GLU) as a promising bacterial candidate for companies like TerraMax Inc. seeking to improve biofertilizer performance, ultimately reducing the need for synthetic fertilizers and promoting environmental sustainability.
Vocal responses in animal behavior are important because they are essential for survival, reproduction, and complex social dynamics. We explored how behavioral responses in Gray Wolves (Canis lupus) vary based on members of the Canidae family and members of the wolf pack. We hypothesized that captive gray wolves would vocally respond when exposed to auditory stimuli of other animals within their phylogenetic tree. We predicted wolves would respond to recordings to signal wolf presence or to establish claims on territory. Our second hypothesis and prediction were that a dominant wolf would initiate a howl, where subordinate members would subsequently howl. We played three playback howls each from five species in the Canidae family. We expected a vocal response to auditory stimuli, as wolves in smaller enclosures were more likely to respond to playback howls. We expected the dominant pair to be leading these behaviors, which explains that alpha breeding pairs influence the behaviors of the pack. Our results showed that there was no statistical significance, as phylogenetic distance and social hierarchy did not influence the proportion of vigilance. This may explain that vocal response is shaped by other factors aside from phylogenetic or social rank.
Antibiotic shortages in tandem with the rise of antibiotic resistance pose significant global public health threats. These obstacles emphasize the importance for continued research and support in the search for novel antibiotic-producing bacteria that carry new lines of defense against human pathogens. Soil contains a biodiverse community of microbial species with an extensive range of antibiotic producing pathways. In nutrient-poor soils, competition amongst microbes is enhanced driving the evolution of unique antimicrobial compounds that may be used in the production of novel antibiotics. This study sought to isolate and characterize antibiotic-producing bacteria from nutrient-poor soils. Soil samples were extracted using a core from three sites. The samples were plated to observe microbial growth. Unique colonies varying in morphology, pigmentation, and size were identified and transferred to a separate plate. The unique colonies were tested for antimicrobial activity against four pathogens: Escherichia coli, Staphylococcus aureus, Bacillus subtilis, and Salmonella typhimurium. Colonies that inhibited the growth of the pathogenic strains were isolated and characterized using the Gram stain procedure. Across the three sites 23 isolates were obtained. The isolates showed resistance to E. coli, S. aureus, and B.subtilis. Further genetic testing will be needed to taxonomically identify novel antibiotic-producing bacteria.
Understanding the effects chemical exposure has on neurodevelopment is crucial for protecting human health and improving environmental policies and safety. Our research uses zebrafish to investigate how environmental and chemical exposures influence early development, as their embryos allow us to observe changes across multiple biological levels. Within our collaborative lab structure, research teams work together to examine how various factors affect development from genes to behavior. My role in the lab includes training in zebrafish-based experimental techniques and beginning to conduct literature analysis to identify candidate chemicals for future experiments.
Methylmercury is a common environmental contaminant in the Great Lakes region of the United States. The glutathione pathway (GSH) involves antioxidant signaling and is responsible for the efflux of toxins such as methylmercury from cells. The object of this study was to verify and characterize mutations in the GSH-associated gene gstp2 in zebrafish. We crossed prospective mutant zebrafish with wildtype zebrafish in an effort to create heterozygous and homozygous mutant zebrafish. To identify mutant zebrafish, we used PCR and restriction enzyme reagents that assess the gstp2 DNA sequence targeted by CRISPR-Cas9 reagents for mutation. Gel electrophoresis was used to visualize the results. This approach was able to identify the presence of mutated gstp2 DNA sequences at the target site. Creation of a homozygous mutant gstp2 zebrafish line will allow for experiments on the influence of gstp2 in embryonic development and developmental toxicity.
In epidemiological studies, prenatal cortisol exposure has been linked to increased autism spectrum disorder (ASD) prevalence. Cortisol is the primary hormone released in the body in response to stress and plays a key role in development. Fragile X syndrome (FXS) is the most common inherited form of intellectual disability, and 30–50% of children with FXS are also diagnosed with ASD. FXS is caused by loss of function mutations in the FMR1 gene on the X chromosome. Our objective was to investigate how embryonic cortisol exposure interacts with FMR1 loss by assessing transcriptome changes in cortisol-treated fmr1 knockout zebrafish embryos. Embryos from wild-type zebrafish and fmr1 knockout zebrafish were exposed to 5uM cortisol solution or vehicle solution at 6 hours post fertilization. Total RNA samples were isolated at 24-, 48-, 72-, and 120-hours post-fertilization (hpf). Sample quality was assessed by spectrophotometry, and RNA concentrations were determined by DNA-binding fluorescent dye methods. Samples will be submitted for RNA-seq analysis to test for genotype-dependent, treatment-dependent, and gene x treatment interactions in terms of neurodevelopmental gene expression across timepoints. These results will help characterize the interaction of embryonic cortisol exposure and FXS genotype in terms of gene expression regulation.
Fragile X Syndrome (FXS) is a genetic condition caused by mutations in the fragile X messenger ribonucleoprotein 1 gene (FMR1) on the X chromosome. FXS is the most common inherited form of intellectual disability causing symptoms such as autism spectrum disorder (ASD). Studies have suggested links between higher prenatal cortisol levels and increased frequency of ASD. The purpose of this study was to compare how embryonic cortisol exposure influences early development in wildtype zebrafish embryos and fmr1 knockout zebrafish embryos at 48 hours post fertilization and 72 hours post fertilization (hpf). Zebrafish embryos were collected and dispersed into petri dishes and treated at 6hpf with cortisol concentrations of 0uM, 10uM, 25uM, or 250uM. At 48 hpf and 72 hpf, embryos were dechorionated and imaged via brightfield microscopy to assess developmental phenotypes. Embryos were analyzed and scored for severity of phenotypes, including pericardial edema, spine curvature, eye size, yolk sac abnormalities, and body size. Survival and phenotype severity were then compared between the genotypes across 48 hpf and 72 hpf. This analysis defines morphological sensitivity to cortisol exposure at different developmental stages. Comparison of wildtype zebrafish embryos to fmr1 knockout zebrafish embryos also provides insight into FXS gene-environment interactions.
Methylmercury (MeHg) toxicity is a notable public health concern in the Great Lakes region. The glutathione pathway (GSH) involves antioxidant signaling and is responsible for the removal of toxins like MeHg from cells. There are many genes involved in the GSH pathway; one of these genes is gclm. Previous lab work has developed a mutant zebrafish line by creating a germline mutation using CRISPR-Cas9 reagents that contains a loss-of-function deletion in the gclm gene. The objective of this study was to create additional gclm mutant fish for investigating developmental MeHg toxicity. Zebrafish of different gclm genotypes were bred to produce heterozygous and homozygous mutant zebrafish. To identify mutant zebrafish, we used PCR and restriction enzyme reagents that target the DNA mutation site in the gclm gene. Results were visualized using gel electrophoresis. The creation of a mutant gclm zebrafish line will allow for future experiments on the influence of gclm in embryonic development and developmental toxicity.
Fragile X syndrome (FXS) is a genetic disorder caused by a mutation of the fragile X messenger ribonucleoprotein 1 (FMR1) gene on the X chromosome. FXS is one of the most common causes of inherited intellectual disability and has a high comorbidity with autism spectrum disorder (ASD). Current research suggests that early embryonic cortisol exposure is associated with increased ASD prevalence, though the molecular mechanisms are still unknown. Our objective was to determine how embryonic cortisol exposure affects the physical development of zebrafish with and without the fmr1 gene. Wildtype zebrafish embryos and fmr1 knockout zebrafish embryos were treated at 6 hours post fertilization (hpf) with cortisol concentrations of 0 μM, 10 μM, 25 μM, or 250 μM. Images were taken and analyzed at 48 and 72 hpf via brightfield microscopy. Embryos were analyzed and separated into different categories based on the severity of morphological phenotypes, including body size, edema, eye size and spine curvature. Comparisons of phenotypes across cortisol concentrations were made within each genotype to assess dose-dependent responses. Morphological severity was then compared between wildtype zebrafish embryos and fmr1 knockout zebrafish embryos to assess genotype-specific differences.
Rare diseases affect 30 million Americans, many of whom remain undiagnosed due to limited functional characterization of DNA variants. Propionic acidemia is caused by variants in PCCA or PCCB that impair enzyme function, leading to severe metabolic dysfunction, often presenting in early infancy. While the Wisconsin newborn screening panel tests for this disorder, screening is neither 100% effective nor does it identify the cause of propionic acidemia in each patient. There are 979 reported DNA variants of uncertain significance or conflicting classification in PCCA and PCCB, meaning that it is unclear if these mutations cause the disease: thus, identification of one of these variants in a patient does not equal clear diagnosis. To address this gap, our lab uses a minigene system to examine whether variants have functional effects. Although we can effectively assess individual variants with this system, it is a relatively low-throughput method. We present our efforts at optimizing this system through improved sample processing, next-generation sequencing (NGS), and development of efficient R scripts. An improved pipeline should accelerate the resolution of variants of uncertain significance associated both with propionic acidemia and across rare genetic diseases.
Over time, biodiversity can change and vary in many ways, for example, in response to urbanization, climate change and invasive species. Pollinators are important animals as they facilitate plant reproduction and maintain plant biodiversity. Our research compares historical data about plant-pollinator relationships in Putnam Park and how it compares to modern data we collected over the summer. We expected to see similar numbers of plant-pollinator relationships, but with some new additions and missing links. To test this hypothesis, we first collected data by observing and catching non-bee pollinators in Putnam Park. We compared the data we collected to the historical data obtained from years of 1882-1928, and this allowed us to see if there was a change in plant-pollinator interactions. We evaluated present and past plant-pollinator relationships by creating a plant-pollinator network to model the consistent and changing relationships. We collected 65 specimens throughout our 10 days of observation between June 1st and September 1st of 2025. We also observed 49 different flowering plant species. We found multiple plant-pollinator relationships that have disappeared, as well as new ones that have appeared, telling us that there are both new additions and missing links that contribute to the biodiversity of Putnam Park.
Bumble bees (Bombus spp.) are among the most important wild pollinators, contributing significantly to ecosystem health and stability. However, due to habitat loss, climate change, and the continuous increase of pesticide use, many species have experienced population declines. Eau Claire, WI, has been reported as home to 15 Bombus spp. including the endangered Rusty Patched Bumble bee (Bombus affinis), making it an important location for bumble bee conservation. This study aims to characterize foraging behavior and preferences of local Bombus species to improve our understanding of preferred food plants to support conservation. To do this, a 12-week field study monitored bumblebee abundance and available flowers at three locations around Eau Claire. By conducting timed field surveys at each location each week and noting flowers visited, we evaluated species presence and abundance and created a flower-Bombus interaction network. Over 177 surveys conducted, 13 different species of Bombus were observed. Each species of Bombus observed had unique peaks in abundance and responses to ambient temperature. 50 different species of flowers were visited by bumble bees over the surveying period (June-September), with the most visited species being Monarda fistulosa. Knowing the bumble bees diet preferences can inform pollinator planting practices in Western WI.
Fragile X syndrome (FXS) is the most common cause of inherited intellectual disability and the leading genetic cause of autism spectrum disorder. FXS is caused by loss-of-function mutations of the FMR1 gene; how FMR1 loss affects neurodevelopment is still being characterized. Our objective was to assess how the FXS genotype affects swimming behavior development in wildtype zebrafish vs fmr1 knockout zebrafish. Zebrafish larvae of wildtype and fmr1 knockout backgrounds were assessed at 5 days post fertilization for distance moved, time spent moving, and velocity over an alternating light-dark cycle using DanioVision hardware with EthoVision XT software (Noldus). RStudio was used for performing Kruskal-Wallis tests to test significant differences between means. Wildtype zebrafish and fmr1 knockout zebrafish had similar swimming patterns in response to alternating light-dark exposure. These results indicate that fmr1 does not regulate swimming behavior development in larval zebrafish.
Fragile X syndrome (FXS) is a genetic disorder caused by loss-of-function mutations of the FMR1 gene and is the most common genetic cause of autism spectrum disorder. Prenatal maternal stress, resulting in increased embryonic cortisol exposure, may also be related to the development of intellectual disability, however, the impact of cortisol on the neurodevelopment of individuals with FXS is not well characterized. To investigate the interaction of cortisol and FXS genotype, wildtype zebrafish embryos and FMR1 knockout zebrafish embryos were treated with cortisol concentrations of 5 µM, 0.5 µM, 0.05 µM, or a vehicle solution between 6 hours post-fertilization (hpf) and 5 days post-fertilization (dpf). At 5dpf, zebrafish were placed in a 96-well plate and evoked swimming velocities were captured across alternating light-dark cycles using a commercial motor tracking system. While zebrafish larvae showed significant difference in light vs dark swimming behaviors, there was no difference in velocity at any cortisol concentration for both AB and fmr1 strains. These results indicate cortisol exposure does not impact swimming behavior development in wildtype or FMR1 knockout zebrafish strains, suggesting no interaction of cortisol and FXS genotype for this phenotype.
A major question in biology involves our understanding of how social connections influence the spread of behavioral innovations in natural populations. In 2024, we documented one such behavioral innovation—the novel emergence of vole hunting in the California ground squirrel (Otospermophilus beecheyi). In the current study, we aimed to explain the mechanisms by which this behavior spread and to use DNA evidence to uncover whether members of our study population consumed other vertebrate prey during the study period. We isolated DNA from 100 fecal samples and used PCR to amplify a 12S local of the vertebrate mitochondrial genome while applying a blocker oligonucleotide to minimize the amplification of ground squirrel DNA. Our genetic data complemented our behavioral observations of vole consumption, and, surprisingly, also revealed the consumption of pocket gophers, salamanders, and three species of mice. Next, we will use network-based diffusion analyses (NBDA) to examine the extent to which the spread of vole consumption is explained by social interactions or shared space use among members of the California ground squirrel study population. Together, our study furthers our understanding of dietary flexibility and highlights the value of molecular tools to inform ecological field studies.
Given rapid urbanization across the globe, coping with humans is an important aspect of life for most animals. Although human presence is often accompanied by human associated predators such as dogs, the effects of humans and dogs on animal physiology are rarely considered together. Here, we leverage over 4,500 measures of fecal glucocorticoid metabolites (FGMs), a biomarker of physiological “stress”, collected from California ground squirrels (Otospermophilus beecheyi) from two sites differing in human activity. Drawing from twelve years of study, we explored the effects of dispersal status, human presence, and dog activity on FGMs in adults and juveniles. Between-site dispersal revealed site-level effects on stress physiology. Whereas FGMs increased after immigrants moved to the more disturbed site, those dispersing to the less disturbed site failed to subsequently reduce FGMs. Responses to human and dog activity were age- and mass- dependent. Whereas juvenile FGMs did not covary with dog activity, juveniles trapped in areas with high human activity had elevated FGMs. In contrast, dogs represented a major stressor for adult animals, but the effects were generally shielded by human presence. These findings uncover complex relationships among anthropogenic factors, stress physiology, and habitat selection in wild animals living in a human-influenced world.
In the context of human-induced rapid environmental change, long-term data on individually recognized social mammals have the potential to offer novel insights into the extent to which animals can cope with social and ecological stressors. Long-term studies are needed to investigate the impact of climate change on the fitness of beavers, and minimally invasive measures of stress physiology offer meaningful biomarkers of these effects in the lives of free-living mammals. Here we present preliminary results of a newly established collaboration with the Norwegian Beaver Project. Glucocorticoids (GC) are steroid hormones that reflect physiological stressors. Because we are extracting GC metabolites from hair samples rather than directly from blood, species-specific validation is required. Thus, we performed an analytical validation for Enzyme-linked Immunosorbent Assays (ELISAs) for (i) cortisol and (ii) corticosterone to identify the predominant hormones in our samples. Our analytical validation will inform our ability to make meaningful connections between hundreds of GC samples and individual-level life-history and habitat measures collected from approximately 25 beaver families since 1997 (i.e., 25 years). Going forward, uncovering the social, ecological, and anthropogenic factors linked to this important biomarker and its associated fitness consequences will advance conservation goals.
In the context of human-induced rapid environmental change, monitoring of wildlife at different sites can offer useful information about how local fauna are responding to human presence. Here we build upon a previous study on the behavioral ecology of North American beavers (Castor canadensis), a native ecosystem engineer, through monitoring using camera traps. We surveyed various public lands and identified several active family groups of beavers in West-Central Wisconsin. Our camera trap data also captured the behavior of other local fauna based on our video analysis. Using Passive Infrared (PIR) sensors, our trail cameras detect captured video footage across each 24-hour cycle including social behaviors of river otters and beavers as well as inquisitive bears and foraging behaviors by squirrels, deer, and native birds. We also detected flying squirrels gliding down from trees at night, and bobcats transversing local habitats. Future research will focus on the extent to which vertebrate assemblages vary across sites and with human activity. The plethora of wildlife we monitored on our camera traps illustrates how beavers play a key role in shaping diverse and healthy wetland ecosystems. Our work has important implications for the management of wetlands and conservation of beaver populations in the region.
The ongoing rise of antibiotic-resistant microbes is a major concern in the medical field. Many pathogenic bacteria that were once vulnerable to common antibiotics have developed resistance, posing significant challenges to medicine and public health. Infections caused by these resistant bacteria are both expensive and challenging to treat. To address this growing crisis, we aim to identify new antibiotic-producing microbes isolated from soil samples. Following the collection of the soil, testing was done to determine the presence of any substances produced that either inhibit or kill any of the tester strains used (Salmonella typhimurium, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis). We have identified eleven strains of microbes that show evidence of producing an antimicrobial substance. We are working to identify these strains and determine if the antimicrobial substance they are producing is novel.
Major food pulses can shift the diets of wildlife over short periods, but little is known about the downstream consequences of foraging decisions on parasite loads. In the current study, we capitalized on our observations of the novel emergence of widespread hunting and carnivory of California voles (Microtus californicus) by marked individual California ground squirrels (Otospermophilus beecheyi) in June and July of 2024. Rodents can be definitive hosts, harboring adult tapeworms in their gut if they ingest tapeworm eggs or intermediate hosts (i.e., infected insects harboring tapeworm eggs). We predicted that sudden dietary shift towards eating other mammals may influence the prevalence of tapeworms in ground squirrels. As part of long-term study on California ground squirrels in the San Fransico Bay Area, we conducted fecal floats to quantify the prevalence of tapeworms in two study populations of ground squirrels before (2023), during (2024), and after (2025) the vole hunting year. Whereas tapeworms were prevalent in squirrel fecal samples before and after the vole boom, tapeworms were rarely present in vole-year samples. Our data is consistent with the notion that dietary shifts can radically influence the prevalence of parasites suggesting that ecological shifts can influence the health of animals.
The North American gray wolf (Canis lupus) is a native keystone predator that contributes to healthy ecosystems across Wisconsin. Despite its broad historical range, anthropogenic development caused friction between pack territories and local farming operations, ultimately leading to the species’ extirpation from the state in the late 1950s. Over the past 25 years, wolf packs have recolonized Wisconsin forests from source populations in Minnesota, and local populations have now reached saturation in the greater Eau Claire region. Our team partnered with the Timber Wolf Information Network (TWIN), an organization established in 1989 to promote wolf awareness and conservation, to support efforts aimed at reducing human-wolf conflict. We monitored wolf activity by identifying tracks in snow and mud. Observations of raised-leg urination at scent posts provided insights into pack boundaries, breeding pair presence, and estrous cycles. Camera traps were deployed within established pack territories to quantify wolf presence and behavior, allowing us to characterize pack dynamics more accurately. These in-depth behavioral surveys will inform responses to population control and maintain beneficial populations of wolves in Wisconsin. More broadly, our project is raising public awareness about the historical impacts of wolves and the ecosystem benefits of their restored presence in Wisconsin forests.
Widespread human impacts – from climate change to unprecedented rates of human visits in once relatively untouched wilderness areas – contribute to rapidly changing selective pressures on wildlife. Long-term studies on marked individuals can offer insights into population and community dynamics over time. The current research aims to reveal the demographic and behavioral patterns of California ground squirrels (Otospermophilus beecheyi) before and after a boom year of California voles (Microtus californicus). Our field study in California has live-trapped, marked, and released individual ground squirrels since 2013 at two study sites. We also directly observed behavioral interactions among squirrels, recorded predator sightings at landmarks, and conducted behavioral assays to assess fear responses to humans. In Summer 2024, we documented the emergence of widespread hunting and consumption of voles by ground squirrels in our two study populations. By Summer 2025, the ground squirrel population crashed, whereas predator numbers soared. We report on these patterns as well as increased fearfulness by squirrels and increased predator sightings, which exceeded numbers documented in previous years. Our findings offer insights into how periods of prosperity (e.g., boom years) and catastrophic turnover events (e.g., demographic crashes) shape wildlife populations that generate ripple effects within ecological communities.
Pollinators are crucial for plant reproduction and diversification. The plant genus Protea of the Cape Floristic Region of South Africa, a global biodiversity hotspot, is an example of how evolutionary radiations can potentially be driven by transitions among primary pollinators. The sprawling shrub Protea venusta has intermediate morphology and is listed as either mammal or bird pollinated, yet has no empirical documentation of pollination. Although it is difficult to document field observations of ground-dwelling mammal pollination, remote motion-activated camera analysis has enhanced our ability to capture 24-hours pollinator activity. This technology allows us to both decipher temporal activity patterns and identify novel pollinators. We deployed six camera traps in a population of Protea venusta at Swartberg Pass in the Western Cape to assess whether this species is predominantly bird pollinated, mammal pollinated, or whether it functions within a mixed pollination network. Preliminary evidence suggests that both birds (such as the Cape sugarbird, Promerops cafer) and rodents (such as the spectacled dormouse, Graphiurus ocularis) visit and likely pollinate this species. Understanding an individual species such as Protea venusta is crucial in providing insight into how unique plant-pollinator networks function, adapt, and persist in one of the world's most biodiverse hotspots.
In the summer of 2025, UW – Eau Claire sent four students to the Galápagos Islands to participate in research internships at the Charles Darwin Research Station. The Galápagos Islands are a highly significant archipelago in the Pacific, renowned for their incredible diversity of wildlife, as well as their contributions to Darwin’s theory of evolution. Presently a hub for scientific research and conservation, students were assigned to various projects at the station, where they worked directly with international scientists to aid in cutting-edge research projects mainly focused on habitat and wildlife conservation. Our poster will feature a discussion of what each of us worked on during our 3 months there, with additional information about how students can get involved.
Overuse of antibiotics in agriculture and healthcare settings results in increased microbial resistance to antibiotics. In order to maintain an advantage in treating disease caused by microorganisms, we must discover and develop new antibiotics and minimize misuse. This in turn creates difficulty in treating diseases that were previously treatable with antibiotics. In previous studies we surveyed 4 different farm soils and discovered many antibiotic-resistant bacteria. In this study we used the same soil samples to determine the incidence of antibiotic-secreting bacteria across WI and MN. We tested resistance to gram-positive and gram-negative bacteria. We discovered 24 isolates that produced antimicrobial effects against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). Out of the 8 isolated strains, we found 4 strains that inhibited both E. coli and S. aureus, 3 strains inhibited only S. aureus, and 1 isolate was only effective against E. coli. Preliminary characterization of the isolates indicates that we have 7 gram-positive rods and 1 gram-negative rod. We are in the process of further characterizing strains and identification of antimicrobial properties.
The glutathione (GSH) pathway is an antioxidant pathway found in humans and zebrafish. This GSH pathway plays a critical role in detoxifying cells from substances such as methylmercury (MeHg), which is a prevalent environmental contaminant in the Great Lakes region. Genetic variations in certain GSH genes have been reported to affect methylmercury detoxification efficiency in adult tissue; however, less is known about these environment-gene interactions during embryonic development. To investigate this context, we are creating knockout zebrafish lines with loss-of-function mutations in the GSH pathway genes gclm and gstp2. Mutations were created by microinjection of CRISPR reagents into 1-cell zebrafish embryos. Offspring of mature zebrafish identified with potential mutation were genotyped to characterize the DNA sequence at the CRISPR target area using PCR + restriction enzyme digestion with gel electrophoresis. The purpose of this project is to confirm loss-of-function mutations and generate homozygous mutant zebrafish lines to study how these genes influence developmental toxicity of contaminants during development.
Aedes aegypti is a significant arbovirus vector responsible for transmitting diseases such as dengue, Zika, and yellow fever, which collectively cause millions of symptomatic cases globally each year. Consequently, extensive research has been conducted to understand the basic biology of these vectors and inform public health initiatives worldwide. The sensory behaviors which allow adult mosquitoes to forage and locate hosts have been thoroughly investigated, while less is known about chemosensory behaviors of the larvae. We conducted behavioral assays using custom-built arenas and an array of high-resolution cameras to evaluate the response of Aedes aegypti larvae exposed to chemosensory gradients. By analyzing poses using deep learning models that allow for precise tracking of individual anatomical features, we confirmed positive chemotaxis in response to food. This platform is now being used to explore larval functional anatomy, specifically those associated with chemosensation and olfaction, and their impact on larva behavior and foraging ability.
Schistosomiasis is a neglected tropical disease caused by Schistosoma blood flukes. Intestinal schistosomiasis is primarily due to Schistosoma mansoni, transmitted by Biomphalaria snails. In mammalian hosts, paired adult parasites reside in the mesenteric vasculature; females lay eggs that traverse the intestinal wall to be excreted by the host, but many become trapped in tissues, especially the liver, eliciting immunopathology. S. mansoni is the major laboratory model, and liver-derived eggs are typically used to maintain life cycles and study miracidia behaviors such as photosensation and chemosensation. However, eggs from liver and intestine differ morphometrically, transcriptomically, and antigenically, which may affect experimental outcomes. To test whether these differences endure after hatching to miracidia, the first larval stage, we compared miracidia hatched from mouse liver- versus intestine-derived eggs, sequencing their transcriptomes and quantifying behaviors using high-resolution tracking. Miracidia transcriptomes were distinguishable by egg source, though only a small subset of genes was differentially expressed. In contrast, basic behavior differed markedly. These distinct behavioral programs highlight egg source as a critical variable in experimental design and interpretation.
The goal of many vaccines is to eventually eradicate a disease by getting enough individuals vaccinated to create herd immunity. For example, in 1980, the World Health Organization (WHO) deemed smallpox eradicated globally. This almost became true for measles in the early 2000, but due to increased rates of vaccine hesitancy, infection rates are on the rise. This research will determine the infection rates of measles before the creation of the measles vaccine, after the public release of the measles vaccine, and after the 2020 COVID pandemic, where cases of vaccine hesitancy started to rise. Measles infection rates will also be compared between different demographics. Based on the results, I will discuss why individuals choose not to get vaccinated and what the rise in vaccine hesitancy will mean for cases of other diseases like polio.
Adenosine A2A receptor (A2AR) antagonists are known to increase hippocampal neurogenesis and goal-directed reward-seeking behaviors. However, the connection between neurogenesis and goal-directed behaviors remained unknown. To study the possible causal relationship, first, we sought to examine whether the A2AR antagonist, istradefylline, which is also recently approved for treating Parkinson’s disease. First, 20 mice were divided into two groups. The first group (10 mice) was injected intraperitoneally (i.p.) daily with istradefylline at 5 mg/kg per day, an hour before behavioral testing. The second group (10 mice) was administered the vehicle. To train the mice toward goal- directed behavior, we used a random ratio (RR) training schedule using a 20% sucrose solution. To evaluate goal-directed or habitual reward-seeking behaviors, we examined nosepoke-based reward-seeking on valued (V) and devalued (DV) conditioning in an extinction test (no reward delivery upon nosepoking) after the training was completed. Goal-directed mice showed a sensitivity to outcome devaluation (stop nosepoking on DV condition). Also, we performed the open field test (OFT) to measure locomotor activity and a Y-maze test to test cognitive function. Our results showed that istradefylline-treated mice exhibited increasing trends of goal-directed behaviors and locomotor activities. Future experiments with additional mice will reveal the role of potentiating goal-directed behaviors and possible correlation with neurogenesis.
Widespread human impacts – from climate change to unprecedented rates of human visits to once relatively untouched wilderness areas – contribute to rapidly changing selective pressures on wildlife. Long-term studies on marked individuals can offer insights into population and community dynamics over time. The current research aims to reveal the demographic and behavioral patterns of California ground squirrels (Otospermophilus beecheyi) before and after a boom year of California voles (Microtus californicus). As part of a long-term field study in California, we have live-trapped, marked, and observed individual ground squirrels since 2013. In Summer 2024, we documented the emergence of widespread hunting and consumption of voles by ground squirrels in our two study populations. Here we document the subsequent crash in squirrel abundance and surge in predator sightings in Summer 2025. We report on these demographic, behavioral, and physiological consequences of the vole boom year using an integrated approach. Our findings offer insights into how periods of prosperity (e.g., boom years) and catastrophic turnover events (e.g., demographic crashes) shape wildlife populations that generate ripple effects within ecological communities.
Background: Human Mesenchymal Stromal Cells (hMSCs) are multi-potent cells that have are known to secrete therapeutic factors that can provide trophic support, immunomodulation, and vascular repair. However, like most cell therapies, using whole hMSCs comes with risks, such immune rejection and tumor formation. We wanted to address this challenge and find a way to improve hMSC therapies to make them safer and more effective. The therapeutic secreted products of hMSCs can be packaged into tiny particles called extracellular vesicles (EVs). We investigated the method of isolating EVs from hMSCs after transfecting them with mRNA, encoding for an anti-inflammatory compound called interleukin 10 (IL-10). Our goals were to determine if mRNA can be used for transient production of IL-10 by hMSCs, and to assess the IL-10 levels expressed freely and the amount being packaged into EVs from the genetically modified cells.
Hypothesis: We hypothesized that hMSCs transfected with IL-10 mRNA will produce more IL-10 free floating in the media and contained within the EVs than the GFP transfected cells.
Methods: To test this, hMSCs were cultured to 80-90% confluency and transfected with IL-10 mRNA (IL10-MSCs) or GFP mRNA (GFP-MSCs) using Lipofectamine for four hours. Cells were then incubated in complete media for one day, followed by serum-free conditioned media for two days. The conditioned media was collected and EVs were isolated using size exclusion chromatography. Nanoparticle tracking analysis was used to confirm successful EV isolation and the amount of IL-10 in whole media or EVs was quantified using ELISA.
Results: Our results confirmed we successfully isolated EVs from conditioned media with ~65% of EVs being in the exosome size range (50nm-150nm) and ~33% in the microvesicle range (100nm-1000nm). The average concentration of EVs was 5.8 x 109 particles/mL. We also demonstrated that IL-10 over secreting hMSCs produced significantly more IL-10 than GFP controls in whole media and showed a trend in increased levels in EVs. IL-10 was detected freely in whole media and in EVs; however, the majority was being secreted freely into the media.
Conclusion: In conclusion, we demonstrated that hMSCs can be transfected with IL-10 mRNA and produce IL-10 freely and also package it into EVs at greater amounts than the control. Future studies can focus on improving the loading efficiency of IL-10 into EVs, through more genetic modifications. The currently mRNA technology tested here is effect for enhancing hMSC cell therapies, with great potential to also be used for cell-free therapy that is safer, more targeted, and potentially more effective for treating many diseases and injuries.
Deep brain stimulation (DBS) of the subthalamic nucleus (STN) and globus pallidus internus (GPi) is an established neuromodulation intervention for the treatment of Parkinson’s Disease (PD). Although highly successful, DBS of these nuclei can produce side effects due to their involvement as part of the complex hyperdirect pathway. The primary motor cortex (M1) projects to the STN and plays a role in PD. Improving characterization of M1 projections through the hyperdirect pathway could present potential new targets for improved selectivity and symptom relief in DBS for PD. In this study, we injected red fluorescent retrobeads into the STN of a healthy rat to retrogradely label its afferent projections. Tissue was sliced in 40 um sections, processed, and analyzed using an immunohistochemistry approach with DAPI staining, imaged at 20x magnification and quantified via fluorescent density. Retrobeads were found in a total of nineteen brain structures. The structures which appeared most often containing retrobeads across all twenty-four images were the ventral posteromedial thalamic nucleus (VPM), ventral posterolateral thalamic nucleus (VPL), zona incerta-ventral (ZIV), subthalamic nucleus (STN), internal capsule (IC), reticular nucleus of the thalamus (Rt), and the primary somatosensory cortex (S1). All other structures containing retrobeads had very limited fluorescence intensity. Contrary to expectations, no retrobead labeling was found in the M1. Additionally, the internal capsule showed the highest retrobead signals, suggesting a critical role in this pathway. The structure with the lowest retrobead signal was the basal nucleus of Meynert. While limited by the sample size, these preliminary findings support the complexity of the hyperdirect pathway and the important role of the internal capsule in the basal ganglia and hyperdirect pathway. Above all, these results highlight the potential of retrobead-based circuit mapping to re ne understanding of the STN’s afferents, ultimately improve DBS targeting strategies for PD.
Multiple sclerosis (MS) is a chronic autoimmune disease that affects millions each year. It causes the body to mistakenly attack its own immune system, causing brain inflammation. One of the ways it does this is through a process called demyelination, which destroys the protective coverings of nerves called myelin sheaths. It is known that inflammation is what triggers demyelination, which is contributing to the progression of MS. Current therapies focus on targeting the inflammatory stage of the disease, but therapies targeting progressive phase of MS are not available. A previous study done in my lab, found that during demyelination, there is an increase of Interferon Stimulated Gene 15 (ISG15) expression in cortical neurons in mice demyelinating models and is elevated in postmortem tissue from the cortex of MS patients suggesting that ISG15 expression, might be a shared stress response in neurons following primary oligodendrocyte loss and inflammatory demyelination. ISG15 is known to be a key contributor to the immune system and is primarily known for its role in modulating immune cells. However, its role in demyelination and neurodegenerative disorders is still unknown. However, we hypothesize that by ablating ISG15 in all cells, as well as specifically in neurons, this will lead to a preservation of NeuN+ cells in the cortex during cuprizone induced demyelination. To test this hypothesis, two groups (A and B) were selected, each with two genotypes of mice in each group: a control, and experimental. In group A, the control group was the B6 wildtype (WT) mice with ISG15 expression in all cell types. The experimental group had the ISG15 conventional knockout (KO) mice group, that did not have ISG15 expression in any of the cell types. In group B, the regular littermate control mice group was Syn Cre-, with ISG15 expression in all cell types. The experimental mice group was the ISG15 conditional knockout (cKO; Syn Cre+ ISG15 fl/fl) group, having ISG15 present in all cell types except neurons; allowing to study its role only in neurons. Experimental design started with inducing demyelination in these mice through a cuprizone diet, starting at 6 weeks old. For the WT and KO mice, at 3, 6 and 12 weeks after starting the cuprizone diet. For the Syn Cre+ and Syn Cre- mice, they were only perfused and collected at 6 weeks after starting the cuprizone diet. Brain slices were stained with a NeuN antibody and counterstained with DAPI. Using the confocal microscope, the stained slices were imaged, and they were quantified for NeuN+ cells in the cortex using stereological sampling parameters. At 12 weeks on a cuprizone diet, preliminary results showed neuron density was preserved in the ISG15 knockout genotype compared to the wildtype control in demyelinating mouse models. In conclusion, our preliminary finding was that ISG15 deficiency protects neurons during demyelination.
Background & Objectives: Spinal cord injury (SCI) abruptly alters the protease microenvironment, including elevations in thrombin capable of activating Protease Activated Receptor 1 (PAR1). PAR1 activation in astrocytes promotes pro-inflammatory and neurotoxic responses that hinder neural repair. Previous studies using PAR1 knockout mice demonstrated that knockout of the PAR1 gene reduces reactive astrogliosis and inflammation while preserving neurons and axonal integrity (Radulovic et al., 2016; Kim et al., 2021). Building on these findings, we employed a thoracic compression SCI model (T8–T9) in adult female C57BL6J mice to test whether pharmacological PAR1 antagonism could similarly sway astrocytes toward pro-repair states and thereby remodel the injury microenvironment.
Methods: The PAR1 antagonist (SCH79797) was delivered following thoracic compression (T8-T9) SCI in adult female C57BL6J mice. Immunophenotyping was completed to quantify astrocyte markers, including GFAP in addition to pro-repair markers (EMP1, S100A10) and a pro-inflammatory marker (C3D).
Results: PAR1 antagonism was associated with a downregulation of astrocytic pro-inflammatory signatures and concomitant upregulation of pro-repair markers.
Conclusion & Implications: Our findings suggest that PAR1 antagonism may promote astrocyte compartment skewing from inflammatory states toward a pro-repair phenotype, which could in turn facilitate neural regeneration. Integrating interventions targeting PAR1 with precise cell-phenotype mapping could unlock new therapeutic pathways to reprogram glial behavior and optimize CNS regeneration after injury.
Acetyl-CoA is a central metabolite crucial for energy production, neuroprotection, and the activity of the citric acid cycle to support oxidative phosphorylation in brain cells. Alzheimer’s disease (AD) is a neurodegenerative disorder that affects more than 7 million Americans characterized by metabolic dysfunction, synaptic degeneration, and alterations in mitochondrial function in early-stage disease mechanisms. One prominent characteristic of the disease is a reduction in glucose uptake, resulting in a diminished level of acetyl-CoA. Along with decreased acetyl-CoA levels, AD also presents reduced histone acetylation, one of the main downstream targets of acetyl-CoA, subsequently resulting in a downregulation of gene transcription. Previous studies demonstrated that mild inhibition of mitochondrial complex I (mtCI) with tricyclic pyrone compound CP2 induces the adaptive stress response, activating multiple neuroprotective mechanisms in brain cells, including increased glucose uptake and utilization. C273 is a small molecule with similar dynamics to that of CP2. It binds to the same site of mtCI as CP2 with differing efficacy and concentration. Optimized for clinical applications, C273 shows greater promise as an alternative therapeutic approach to treating AD. The aim of this project is to assess whether the inhibition of mtCI with C273 affect to the abundance of acetyl-CoA and improves transcription of genes involved in cognition. Neuroblastoma SH-SY5Y cells that express mutant human APP protein (APPswe) and control cells were treated with C273 (50 nM or 500 nM) and CP2 (2.5 μM) for 24 hours. Mass spectrometry was used to measure the acetyl-coA level. The change in acetyl-CoA–sensitive targets (H3K27 acetylation and total histone H3) were analyzed by Western blot. At 24 hours of treatment with C273 or CP2, acetyl-coA levels increased significantly in both control and APPswe cells. Confirmation of acetyl-CoA level changes by assessing downstream effects on H3K27 acetylation. Importantly, C273 induced a similar percentage increase acetyl-coA as CP2, but at a 50-fold-lower concentration. These findings suggest that C273-dependent activation of multiple mechanisms essential for improved energy homeostasis in APPswe cells results in an increase of whole cell acetyl-CoA levels enhanced gene expression associated with cognition via histone acetylation. These preliminary results will be validated in future studies.
