2020 Abstracts

Allen, Brian; Covey, Tracy; Davies, Don; Eccles, Nick; Farnsworth, Brian; Ferguson, Parker; Hart, Sierra; Lowder, Jordan (Weber State University)
Faculty Advisor: Davies, Don (Weber State University, Chemistry and Biochemistry); Covey, Tracy (Weber State University, Chemistry and Biochemsitry)

Chalcones refer to biological molecules with the structure trans 1,3-diphenylprop-2-en-1-one. Biological chalcones and chalcone derivatives display anti-tumor, anti-fungal, anti-inflammatory and antibiotic properties. To understand the role of the chalcone structure in tumor cessation, derivatives to the original chalcone were synthesized using aldol condensation reactions. HeLa and HEK-293 cells were treated with the synthesized chalcone and an LD50, or the concentration of chalcone required to kill half of the cells, was calculated. The LD50 was then used to determine the efficiency of the chalcone derivative. Correlations between the structure and activity suggest that a Michael reaction occurs at the cell and indicate that that an aromatic ring at C3 is likely necessary. Further research will help determine the structures of more cytotoxic compounds.

Wasden, Kayla; Suisse, David; Kaundal, Amita (faculty mentor) (Utah State University)
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)

Many plants secrete substances to create a more favorable environment, including chemicals that kill pathogenic microbes or competing plants. Artemisia tridentata, also known as "Big Sagebrush," is prevalent in the Rocky Mountain region of the United States and is known to have antimicrobial capabilities. We will study the potential antimicrobial activity of Artemisia tridentata.

Studies report that chemicals released by the leaves and branches of A. tridentata affect bacteria native to deer rumen. Another study showed that 27 actinomycetes (anaerobic bacteria that form colonies) strains found in the rhizosphere of A. tridentata demonstrated antibacterial activities when tested on E. coli, Bacillus subtilis and Staphylococcus aureus. Native Americans traditionally used A. tridentata to relieve stomach pain, colds, coughs, sore eyes, snake bites and as an insect repellent. Researchers found several compounds, including flavonoids, that can affect antimicrobial activity. Articles regarding antimicrobial activities in A. tridentata were published between 1967 and 2004. With the chronological gaps and considering the progress that biological and molecular technology has made in recent years, knowledge of the chemicals released by A. tridentata lies largely untapped. In this study, we will investigate the antimicrobial activities of the leaves, stem, roots, and flowers of A. tridentata initially by the agar well diffusion method and followed by validating with the agar disk diffusion method. We will check the antimicrobial activity of the extract from different plant parts of A. tridentata on common bacteria such as E. coli, Bacillus subtilis, and some Pseudomonas spp. of plant pathogens.

The knowledge obtained from this research will further help in the identification and characterization of the secondary metabolites or chemicals involved in antimicrobial activity of sagebrush. Medicinal plants provide a healthy, natural alternative to conventional medication, and may lead to new insights on antibiotics and pharmaceuticals. Besides, Artemisia tridentata is a plant native to Utah and Idaho. It grows everywhere in the surrounding area, making it inexpensive (free) to produce.

Baptista, Gabriela; Payne, Andrew; Obray, J Daniel; Yorgason, Jordan; Weber, K Scott; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences, Psychology)

Cluster of differentiation 5 (CD5) is expressed in both T and B cells. CD5 has been found to display an altered expression profile following chronic ethanol use and during ethanol withdrawal. Specifically, the number of CD5+ B cells is reduced during withdrawal while the number of T cells is increased. Given the apparent sensitivity of these cells to ethanol and recent research suggesting that some ethanol effects are accounted for by neuroimmune interactions we assessed drinking behavior and ethanol induced sedation in CD5 knockout (KO) mice. We found that CD5 KO mice display decreased ethanol consumption as compared with wild-type controls and that ethanol consumption does not increase with repeated exposure in CD5 KO mice. Additionally, CD5 KO mice displayed considerable resistance to the sedating effects of ethanol. Further studies are underway to assess whether there are baseline differences in dopamine dynamics within the mesolimbic pathway between CD5 KO mice and wild-type controls as well as to whether neurons in the mesolimbic pathway differ in their response to ethanol in CD5 KO mice.

Bartlett, Ashley; Phatak, Sumira; Hintze, Korry; Benninghoff, Abby (Utah State University)
Faculty Advisor: Benninghoff, Abby (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)

The gut microbiome modulates various physiological functions related to cancer development including inflammation, cell proliferation, apoptosis, and angiogenesis. Patients with inflammatory bowel disease have a microbiome distinct from healthy controls with consistent observations of reduced gut biomass, decreased diversity within the community, and altered relative abundance. Although a consensus cancer-related microbiome has not been identified, several pathogenic species play an instrumental role in the progression of colitis and tumorigenesis, including: Streptococcus bovis, Helicobacter pylori, Enterococcus faecalis, Clostridium septicum, and Escherichia coli. Gut microbial composition is highly responsive to diet and inadequate intake of micronutrients is a critical feature of the Western dietary pattern. Gut dysbiosis has been proposed to further limit mineral uptake and impair vitamin synthesis, predisposing the host to micronutrient deficiency. Dietary bioactives, such as those in green tea, may function as a mediator between the gut microbiome and basal diet to ultimately prevent colitis associated colorectal cancer (CAC). The overarching objective of our work is to determine the impact of ancestral or multi-generational consumption of the total Western diet (TWD) in a murine model of CAC. Our previous work is the first to investigate how diet induced transgenerational inheritance affects CAC outcome. Our data suggested that multigenerational patterns of exposure to the TWD altered both phenotype and gene expression in third generation offspring. Supplementation with green tea appeared to be most promising after consumption of TWD for multiple generations. Considering that gut microbes are inherited maternally after colonization during vaginal birth, the gut microbiome is a missing piece in this disease model puzzle. The hypothesis of our current project is to investigate whether intake of TWD influences the transmission of microbes and whether CAC outcome is reflected by altered gut microbial composition. Based on other work, we expect the healthy control to possess an abundance of varied bacterial taxa that maintain protective epithelial barrier function and overall homeostasis. On the other hand, a high fat diet would promote increased intestinal permeability, a substantial shift at the phyla level, and increased production of pro-inflammatory cytokines. After TWD consumption, we expect an overall negative phenotypic outcome within the gut microbiome, that includes a breakdown of the epithelial barrier and introduction of pathogenic bacteria. These harmful bacteria tend to thrive on simple sugars that are common in the Western dietary pattern and tend to produce metabolites known as endotoxins that promote dysbiosis.

Anderson, Lars; Baldwin, Haley; Christensen, Ben; Walker, Austen (Brigham Young University)
Faculty Advisor: Griffen, Blaine (Brigham Young University, Life Sciences)

This research looks at the blue coloration on uca pugnax crab carapace above the mouth and between the eyestalks and associates the coloration to the behavior, sexual maturity, and size of the crab, as well as the detection of metals in their environment. Up to ten crabs were photographed within twenty five isolated sites with the objective of gathering a high range of color difference among the uca pugnax. The photos of the crabs were set to match the same scale of light and RGB as to not have interference from external factors such as sunlight or overcast weather. The shade of blue on the carapace provides information about the surrounding environment where the uca pugnax are found.

Sagers, Rachel; Nguyen, Andy; Weston, Jake; Grooms, Nicholas; Eggleston, Morgan; Martin, Randy; Coulombe, Roger (Utah State University)
Faculty Advisor: Coulombe, Roger (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)

The scenic mountain views of Cache Valley in Northern Utah stand in stark contrast with the valley's high concentrations of fine particulate air pollution (PM2.5), some of the worst reported in the United States. The unique geography promotes formation of ammonium nitrate (NH4NO3) from nitrogen oxides produced by motor vehicles and ammonia from dairy cow excreta. Winter atmospheric inversions, exacerbated by the mountainous terrain, trap and concentrate air pollutants. Epidemiological studies have revealed an association between PM exposure and early all-cause mortality. Exposure to PM2.5 is also associated with a variety of cardiovascular, cardiopulmonary, and neurodegenerative diseases, including myocardial infarction, stroke, COPD, lung cancer, Alzheimer's disease, and Parkinson's disease. Previous studies have shown that Cache Valley PM (CVPM) has pro-inflammatory effects, which has been linked to enhanced activation of Akt in human pulmonary epithelial cells. This research examined the cellular responses of human lung (BEAS-2B) cells exposed to CVPM and diesel exhaust particles (DEP), at 1 and 12 µg/ml concentrations of each particle type for a 24 hour exposure period. The CVPM used was collected onto stainless steel plates by a Tisch impactor. Assessment by the comet assay reveal genetic damage to CVPM exposed cells with equal potency to DEP exposed cells. Flow cytometry (p < 0.05) showed CVPM exposed cells had a significant increase in the number of actively-dividing cells compared to control cells. Whole-genome microarray identified affected genes related to inflammatory pathways, as well as activated Akt-dependent pathways. Subsequent qRT-PCR showed that CVPM exposure significantly increased expression of inflammatory markers, including IL-6, CD40LG, PLAG27, and cytochrome P450 (CYP) 1A1 (p < 0.05). Immunoblotting confirmed activation of Akt by phosphorylation of Thr308 in both CVPM and DEP exposed cells. This data supports the hypothesis that CVPM may induce pro-carcinogenic pathways with potency similar to DEP.

Littlefield, Connor; Tessem, Jeffery (Brigham Young University)
Faculty Advisor: Tessem, Jeffery (Brigham Young University, NDFS)

The transcription factor Nkx6.1 is essential for beta cell growth and function. Given that Nkx6.1 is expressed in beta cells undergoing high level expansion, our lab demonstrated that Nkx6.1 overexpression in primary rat islets was sufficient to induce beta cell proliferation and enhance glucose stimulated insulin secretion. However, while these phenotypes are evident in islets from young animals, islets from aged animals fail to induce proliferation or increased insulin secretion. One reason for why Nkx6.1 fails to drive proliferation or increase insulin secretion is due to lost binding partners that allow it to control gene transcription. We hypothesize that loss of Nkx6.1 binding partners curtails its ability to induce gene transcription that leads to proliferation and enhanced glucose stimulated insulin secretion. To test this hypothesis we have used Nkx6.1 BioID to define by mass spectrometry the proteins that interact with Nkx6.1 Here we define three novel interactors, Mef2D, Sirt7, PDX1. This finding will provide us with a greater understanding of Nkx6.1 function in the beta cell, provide us with new gene targets essential for Nkx6.1 function, and allow us to begin to apply these findings to aged beta cells.

Wood, Branzen; Oberg, Taylor; Culumber, Michele; Oberg, Craig (Weber State University)
Faculty Advisor: Oberg, Taylor (Utah State University, Nutrition and Food Science); Culumber, Michele (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology)

The unique flavorings and textures of Cheddar cheese are produced by the degradation of the major milk proteins. One of those proteins, casein, is degraded by the enzyme chymosin and a series of peptidases produced by the starter Lactococcus added to the milk. As casein is degraded, several small peptides accumulate. One of these peptides, ß-casein, can have an adverse bitter taste that is non-desirable and considered a defect in Cheddar cheese. The two main starter cultures used industrially in Cheddar cheese making are Lactococcus lactis subsp. lactis and L. lactis subsp. cremoris. L. lactis subsp. cremoris has been used traditionally in Cheddar cheese making, however, L. lactis subsp. lactis ferments more quickly and is becoming more popular in the cheese industry. With the transition creameries have seen a sharp rise in bitterness during production. Our hypothesis was that while closely related, cremoris synthesizes some peptidases that help with ß-casein degradation that lactis does not. Peptidases found in cremoris include PrtP I and II, Pep X, Pep C, Pep A, Pep T, Pep Q, Pep N, Pep V among others. We searched the genomes of both strains using RAST bioinformatic software, and the databases NCBI and UniProt. The peptidases common in cremoris were also found in lactis. We are now trying to determine if the location of the peptidases on the genomes change how they are regulated or produced. Further, we will begin looking into the genome for other, novel, enzymes that might have peptidase activity that influence bitterness.

Nanasi Sekona, Ashley Balderrama, Carlos Nunez, Kyle Hendricks, Tyson Hillock, and Dr. Olga Kopp (Utah Valley University)
Faculty Advisor: Kopp, Olga (Utah Valley University, Biology)

Over the last 30 years, the incidence of fungal infections has gradually increased. Mucormycosis is a fungal infection primarily caused by Rhizopus Oryzae. The majority of patients who develop invasive mucormycosis die within 12 weeks of diagnosis. Mucormycosis is commonly treated with an antifungal agent called Amphotericin B (AMB). When used in high concentrations, AMB causes severe side effects such as nephrotoxicity. It has been reported that 99% of microbes exist as biofilm: thus, there is a direct association between mucormycosis and biofilms. Shockwave has been shown to inhibit living bacteria in biofilm, but few studies have focused on the effects of shockwave on fungal biofilm. Previous work in our lab showed that shockwaves were effective in damaging biofilms of R. oryzae; but at the same time helped promote the metabolism of surviving R. oryzae. This study aims to investigate the proteins expressed in fungal biofilms when introduced to different intensities of shockwave coupled with the treatment of AMB. This will be accomplished by culturing sporangiospores and propagating R. oryzae biofilms. Standardized biofilm will be treated with 0.5 µg/mL AMB in 1% DMSO, and/or shockwave treatment of 300 pulses at 0.19 mJ/mm2 energy density to be measured against a control group. The proteins will be extracted, determined by 2D gel electrophoresis, and identified by mass spectrometry. Studying protein expression resulting from combination therapy of extracorporeal shockwave and AMB on R. oryzae biofilm could progress research surrounding the difficulties of mucormycosis treatments. Particularly, research aimed at counteracting the antifungal and antimicrobial resistance contributed by proteins in the fungi's biofilm.

Jones, Abigail; Hevel, Joan (Utah State University)
Faculty Advisor: Hevel, Joan (College of Science, Chemistry and Biochemistry Department)

PRMT1 is one of nine known mammalian Protein Arginine Methyltransferases (PRMTs) whose function are to transfer methyl groups from S-adenosyl methionine (SAM) to arginine residues of specific proteins. PRMT1 is known to methylate many different proteins in cells, but the mechanism of target recognition and binding is still unknown. Correct regulation of PRMT1 is critical to proper cellular function; thus, the action of PRMT1 is important to understand. In this study, we seek to elucidate how PRMT1 recognizes and binds its targets by identifying protein substrates of PRMT1 that form a stable complex with the enzyme. Such a protein would allow for additional studies (e.g. crystallographic or cryo-EM studies) to help visualize PRMT1-substrate interactions. Two substrates of PRMT1, TWIST1 and Smad6, have been purified, and the binding affinity of each to PRMT1 has been qualitatively assessed via pull-down assay and Western blot. Ligation-independent-cloning has been used to clone each substrate gene out of a GST-tagged vector and into a His-tagged vector, which will allow for further experiments assessing the stoichiometry of PRMT1-substrate binding.

Hirschi, Blake; Pickett, Brad; Thompson, Jared; Telford, Mady; Berges, Bradford (Brigham Young University)
Faculty Advisor: Berges, Brad (Life Sciences, Microbiology and Molecular Biology)

Staphylococcus aureus (S. aureus) is a commensal bacterium commonly found amongst livestock and near 30% of humans' nostrils. However, through acquisition of certain genes S. aureus may develop antibiotic resistance such as in methicillin-resistant Staphylococcus aureus (MRSA). One hypothesized component lending to acquisition of genetic resistance in S. aureus is the synthesis of colony biofilms. Biofilms are comprised of a variety of substances including secreted polysaccharides, protein and even extracellular DNA. Our work postulates that extracellular DNA-based biofilms will transfer genes for antibiotic resistance at a higher rate than in polysaccharide/protein biofilms. Through employment of polymerase chain reaction (PCR), we aim to characterize a wide sample of methicillin-susceptible S. aureus (MSSA) human associated strains and MRSA livestock associated strains for multiple antibiotic resistances. Co-inoculating pairs of human associated and livestock associated strains, each lacking the other's resistance genes, will provide an environment wherein biofilm-mediated gene transfer may occur. Further pairing based on biofilm composition (DNA or polysaccharide/protein) will yield data concerning which biofilm facilitates gene transfer more efficiently. Subsequent genotyping will confirm whether resulting isolates acquired new antibiotic resistance through biofilm-mediated transfer, thus increasing pathogenicity.

Vilela, Ernie; Seal, Isaac; Ogden, Heath T. (Utah Valley University)
Faculty Advisor: Ogden, Thomas (College of Science, Biology Deapartment)

We are interested in using transcriptome data, generated with next generation sequencing technology, to investigate the evolutionary trends of specific genes and their associated expression in mayflies. We generated an additional transcriptome for mayflies. RNA was extracted from a freshly frozen specimen preserved in RNAlater® (Ambion) using TRIzol® Reagent (Ambion) and cDNA libraries were prepared from mRNA. RNA-seq data was generated using a paired-end protocol (PE100) on Illumina HiSeq2000 with an expected 60 million reads. In order to effectively investigate the large amount of sequences, we created a bioinformatics workflow to analyze the newly generated transcriptome data along with previous data for mayflies. The workflow consists of these main steps: Trinity (Assemblying the transcripts), Transdecoder (Identifying candidate coding regions), HMMER (Searching biological sequence databases for homologous sequences). We tested the workflow looking at opsin genes.

Forbush, Micah; Gold, Roger (Southern Utah University)
Faculty Advisor: Gold, Roger (Southern Utah University, Biology)

The health of a horse's teeth and oral cavity is strongly correlated with the horse's overall health and well-being. As a horse ages the oral cavity undergoes many distinct physical changes, which may lead to changes in the composition of the oral microbiome as well. While there has been extensive research performed on the oral microbiomes of healthy horses compared to horses with various oral diseases, very little is known about normal changes to the horse oral microbiome as they age. The aim of this study was to use high-throughput sequencing to compare the oral microbiomes of horses in different age categories. Total genomic DNA was isolated from oral swabs taken from horses in the 7 day-old, 1-7 years old, 8-15 years old and 16+ years old age categories. The V3-V4- region of the 16S rRNA gene was amplified by PCR and amplicons were submitted for paired-end sequencing on the Illumina HiSeq system. Sample reads were analyzed using the QIIME 2.0 microbiome bioinformatics platform and overall bacterial diversity was compared among age classes.

Terreros, Eduardo N.; Dr. Roberts, Andrew G. (University of Utah)
Faculty Advisor: Roberts, Andrew G. (University of Utah, Chemistry)

Chromatography is an indispensable tool in most chemistry laboratories, as it provides a way to separate multiple components within a chemical mixture. The separation can be evaluated by the use of various chromatographic methodologies that have been developed to target specific chemical properties (e.g. size, mass, molecular interactions) of the desired compounds. Chromatographic methods are utilized to gather qualitative information in order to observe more general trends, or quantitative data to further explore observations in greater detail.

Thin Layer Chromatography (TLC) is a well-established method for collecting qualitative data from chemical reactions. Although Quantitative TLC (QTLC) methods have been developed, they are limited due to their high complexity, and cost prohibitive nature. The advancement of a Semi-Quantitative Thin Layer Chromatography (S-QTLC) method will provide a quick, low cost method to track organic reactions. We developed a low-cost alternative that uses readily available materials to gather images of TLC plates and process them into semi-quantitative data. In order to achieve this, we have constructed a small image gathering box made of opaque acrylic fitted with two 254 nm UV lamps and a Raspberry Pi Zero W with a camera module to acquire the images. The images are imported to a PC and processed by a custom built MatLab function. We demonstrated that this system can be utilized to gather qualitative data. Current studies will validate semi-quantitative data collection and reaction tracking and work is currently being done to validate semi-quantitative data collection.

Fazio, Nicholas; Russell, Michael; Krapohl, John; Andrus, Brayden; Hansen, Marc (Brigham Young University)
Faculty Advisor: Hansen, Marc (Brigham Young University, Physiology and Developmental Biology)

PIM 3 is a proto-oncogene with serine/threonine kinase activity that can prevent apoptosis, promote cell survival and protein translation. Abnormal PIM3 activity contributes to tumorigenesis by phosphorylation of targets that release anti-apoptotic proteins. Pathological PIM3 expression is common in pancreatic and prostate cancer. Inhibiting this kinase activity can be used to therapeutically suppress uncontrolled cell growth in cancerous tissues. Synthetic inhibitors are being developed as therapeutics to treat PIM3 related disorders. Compounds derived from plants and natural sources have therapeutically-relevant biological activity. Additionally, they often well tolerated, making them important starting points for drug discovery efforts. A less widely used approach to discover the biological activity of molecules is built around using a large scale in-silico molecular screening, which has emerged as a critical drug discovery tool. Here, we screen a large (>100,000 compound) virtual library of natural product compounds for binding in the PIM3 ATP binding site, then validate compounds with using cell-based and immuno-based assays. This approach reveals PIM3 inhibition by a saponin scaffold, which suggests potential utility as a therapeutic or as a lead for further optimization.

Leininger, Sara; Minteer, Shelley; Rhodes, Zayn; Sigman, Matt; Pancoast, Adam (University of Utah)
Faculty Advisor: Minteer, Shelley (University of Utah College of Science, Chemisty)

In the effort to improve renewable energy as a response to the depletion of fossil fuels, one important aspect to consider is the availability of such sources. The supply of solar and wind power, for example, faces issues with intermittency. Therefore, it is crucial to develop reliable energy storage methods, with redox flow batteries (RFBs) being of particular interest given their potential low cost and high efficiency. RFBs operate similarly to conventional batteries, except the anode and cathode materials are dissolved in electrolyte solutions, and pumped into the electrochemical cell from external storage tanks. Within the cell, the anode and cathode species are separated by a membrane to prevent them from mixing, which would cause the battery to self-discharge. RFBs can utilize aqueous- or organic-based electrolyte solutions, with organic solvents being especially appealing, as the electrochemical potential window is larger than water. However, one major impediment of using organic solvent is the high chemical crossover rate of anode and cathode species through the membrane, causing rapid capacity fade of the battery. Several research studies have shown that the use of redox active polymers (RAPs) with high molecular weights, paired with a size-exclusion membrane effectively counteracts this problem. The resulting steric hindrance between the small pores of the membrane and these large molecules blocks any crossover from the active species. This study will include the construction of an RFB using two previously developed RAPs demonstrated to have high electrochemical cycling stability as electrolytes. By using RAPs as both anode and cathode materials, it is expected that chemical crossover will be minimized, and the lifetime of the battery will be elongated compared to an RFB with one or both species in monomeric form. This study will be significant in the advancement of RFBs, potentially leading to their widespread use for energy storage.

Marshman, Evan; Peterson, Samara; Call, Gerald; Chaston, John (Brigham Young University)
Faculty Advisor: Chaston, John (Life Science, Plant and Wildlife Science)

In recent years the association between the human gut microbiome and the brain has become a promising field of study. Often referred to as the "gut-brain axis", this connection has greatly enriched our scientific understanding of many disorders that affect the brain and nervous system. A recent study showed the differing richness of bacteria in the microbiota of Parkinson's patients and healthy control subjects. Because recent research shows this connection, we predicted that we would detect variation in the microbiota of D. melanogaster (fruit flies) models of Parkinson's disease, relative to wild type flies. To test this hypothesis, I analyzed 16s rRNA sequence data, reporting the microbiota composition in flies that are a model of Parkinson's Disease, as well as wild type flies. I found one strain of the genus Acetobacter that was differentially abundant between the two fly types. Therefore, for my CURA I will extend my analysis by performing similar analyses by taking a larger set of Parkinson's fly models. Once they are sequenced, I will use QIIME, the same software I used in my preliminary analyses to further our understanding of the taxonomic differences between the gut bacteria of Parkinson's models and wild type flies.

Murley, Jordan; Stockard, Alyssa; Payne, Andrew; Steffensen, Scott (Brigham Young University)
Faculty Advisor: Steffensen, Scott (Family, Home, and Social Sciences; Psychology)

Brain-derived neurotrophic factor (BDNF) is implicated in varied physiological processes. Its main receptor in the central nervous system is tyrosine receptor kinase B (TrkB), and the main ligand for TrkB in the central nervous system is BDNF. It has been shown previously that activating TrkB can cause a downregulation of the chloride-exporting potassium chloride cotransporter 2 (KCC2), presumably resulting in a decreased chloride gradient. It is hypothesized that this down-regulation, caused by an increase in BDNF levels, creates hyperexcitable GABA neurons in the ventral tegmental area (VTA) due to a reduced efficacy of inhibitory currents. Here we investigate this mechanism of adaptation in the context of chronic alcohol exposure. We demonstrate that BDNF levels are elevated in the VTA during withdrawal from chronic alcohol exposure. We also observe that blocking TrkB activity decreases alcohol seeking behavior. Further, we investigate the expression patterns of KCC2 in connection with chronic alcohol administration. Additional work is underway to validate this mechanism and further elucidate its putative role in alcohol dependence.