Robert B. Glassman Memorial Brain, Mind, and Behavior Symposium

The 2023 Glassman Symposium will be held November 2–3.

The Glassman Symposium Poster Session seen from a balcony above

Honoring Outstanding Student and Faculty, and Alumni Research in Behavioral and Life Sciences

The Brain Awareness Week Faculty/Student Symposium was renamed the Robert B. Glassman Memorial Brain, Mind, and Behavior Symposium in 2013 in honor of the late Professor of Psychology Robert Glassman, who played a leading role in developing Lake Forest’s popular neuroscience major.

The symposium consists of a series of talks given by Lake Forest College faculty and alumni on November 2 and an poster session where Lake Forest students and alumni will present their original research on November 3. 

November 2 – Faculty/Alumni Talks

7–9 P.M., Tarble Room, Brown Hall 

7:00 P.M. Opening Remarks, Nu Rho Psi

Nora McLeanNora McLean, PhD

Assistant Professor of Psychology and Neuroscience, Lake Forest College

Beyond Cortisol: Moderators of Psychosocial Stress Reactivity

7:05 P.M.

Britany AvontsBrittany Avonts '17, PhD candidate

PhD candidate in Neuroscience, Rush University

Predicting Pain After Spinal Cord Injury

7:25 P.M.

Stephanie CaparelliStephanie Caparelli '04, JD

Chair of Legal Studies, Assistant Professor of Politics, Lake Forest College

The Dubious Science Behind Shaken Baby Syndrome and Its Dangerous Implications in the Courtroom

7:45 P.M.

Kayla HuberKayla Huber '16

PhD candidate in Psychology, University of Minnesota

Beyond Catcalls: Reassessing the Measurement and Mental Health Correlates of Street Harassment

8:05 P.M.

Steve McGaraughtySteve McGaraughty, PhD

Lecturer in Neuroscience, Lake Forest College, and Director, AbbVie labs

Finding Pain Relief

8:35 P.M.

8:55 P.M. Closing Remarks, Nu Rho Psi

November 3 – Undergraduate and Alumni Research Poster Session

4:30–6:30 p.m., Calvin Durand Hall, Mohr Student Center

4:30 P.M. Chicago Society for Neuroscience Reception and Unveiling of Annual Neuroscience Sculpture

4:45 - 6:10 P.M. Poster Viewing
Enjoy posters and exhibits by current students and recent alumni of original student/faculty research conducted at Lake Forest, Rosalind Franklin University of Medicine and Science, Midwestern University, Northwestern University, University of Minnesota, Illinois Institute of Technology, and Rush University

6:10 P.M. Closing Ceremony
Introduction,  Nu Rho Psi
Remarks, Tara Natarajan, Krebs Provost/Dean of Faculty
Recognition of student scholars and celebratory photo

2023 Student and Alumni Poster Abstracts

Alumni Abstracts

Sleep, Diet and Physical Activity Patterns among Latinx 2-to-5 year olds

Helena Blumenau ‘23, Amelia Bailey, and Augustine

Kang Center for Alcohol and Addiction Studies, Brown University School of Public Health, Providence, RI, USA

The early years of life lay the foundation for physical, cognitive, and emotional development, making them a crucial period to establish healthy lifestyle habits that extend well into adulthood. Among Latinx children (ages 2-5), there is a growing concern regarding potential disparities in sleep, diet and physical activity patterns when compared to their non-Latinx peers. Investigating these factors is crucial as they play a key role in shaping a child’s growth trajectory and overall wellbeing. This qualitative study aims to assess the relationship between diet, physical activity and sleeping habits among Latinx children. Results: We identified five major themes describing the interactions between sleep, diet and physical activity: (1) Diet affects sleep; (2) Play affects sleep; (3) Sleep affects diet preferences and physical activity; (4) Role of the parent and (5) Individualized factors. Within each of these themes, parents identified specific interactions such as poor sleep quality negatively impacts the child’s temperament, eating specific cultural foods helps the child to sleep, and physical activity increases overall sleep duration. Discussion: This study sheds light on the sleep, diet and physical activity patterns of Latinx preschoolers, highlighting the need for culturally sensitive interventions to promote healthy behaviors.

Regulation of Neuronal Inflammation in an Experimental Autoimmune Encephalomyelitis Model via LAG3 Under-Expression

Allison Coffell ‘23, Joseph Reynolds, Kristen Patterson

Center for Cancer Cell Biology and Immunology, Rosalind Franklin University of Medicine and Science, IL, 60064, USA

Multiple Sclerosis (MS) is an autoimmune T-Cell mediated disease of the Central Nervous System (CNS) resulting in degeneration of healthy myelin. The immune checkpoint receptor, Lymphocyte-Activation Gene 3 (LAG3) is typically under-expressed in MS patients. This study elucidates a therapeutic potential of a LAG3-targeting antibody in reducing neurodegenerative virulence in an Experimental Autoimmune Encephalomyelitis (EAE) mouse model. LAG3 antibody-treated mice exhibit decreased levels of proinflammatory Interleukin-17A (IL17a) CD4 presenting T-cells within the brain as well as increased levels of LAG3 in the spleen. Scoring data revealed a slowed progression of symptoms in EAE mice treated with a LAG3 antibody. A transfer experiment revealed the preventative effect of shRNA silencing of LAG3 on EAE pathogenesis. These findings thus demonstrate a role of LAG3 deregulation in reducing neuronal inflammation and altering typical trajectory of MS symptomology in an EAE model.

Targeting MeCP2-Associated Disorders with miRNA Site-Blocking Oligonucleotides

A.M. Vanderplow1, 2 , G.E. Dodis ‘211, 2 , J.J. Cikowski1 , R.G. Gogliotti1, 2

1 Department of Pharmacology, Loyola University Chicago, 2 Stritch School of Medicine, Loyola University Chicago

Rett syndrome (RTT) is a neurodevelopmental disorder caused by loss-of-function mutations in the methyl CpG binding protein 2 (MeCP2), a crucial methyl-reader protein. Despite extensive research spanning nearly two decades, progress in developing effective therapies for RTT has been limited. However, seminal studies have demonstrated that RTT can be reversed in mouse models by restoring MeCP2 levels. While gene therapy appears to be a promising approach for RTT at first glance; MeCP2 is a dose-sensitive gene, and even a 1-fold increase in expression could result in a related neurodevelopmental disorder known as MeCP2 duplication syndrome (MDS). This poses a significant practical challenge, as achieving efficient delivery of MeCP2 to the entire human brain is necessary while ensuring that each cell receives a precise and relatively small amount of the protein. The 3'untranslated region (3'UTR) plays a critical role in the post-transcriptional regulation of mRNA, where numerous microRNAs (miRNAs) and RNA binding proteins collectively contribute to fine-tuning gene expression. The impact of each miRNA on protein levels is generally modest, making strategies that disrupt miRNA regulation of the 3'UTR potentially valuable tools for disorders with a narrow therapeutic window for the target gene. The repressive role of miRNAs on MeCP2 expression has been shown at multiple sites and when overexpressed and this strategy can reverse MDS symptoms in mice. However, its therapeutic potential in RTT remains understudied. We have developed an innovative approach that utilizes a series of locked nucleic acid site-blocking antisense oligonucleotides (sbASOs). These sbASOs are specifically designed to “out-compete” with endogenous miRNAs for binding to MeCP2’s 3'UTR. This strategy effectively enhances or releases the repression of MeCP2 expression in a controlled manner, capping it at sub-toxic levels. We believe that this strategy could be a promising intervention for patients with MeCP2 mutations characterized by missense or latetruncating variants, as the increase of MeCP2 can compensate for the partial loss of function. Our initial data demonstrates that targeting microRNA binding sites using sbASOs can effectively elevate MeCP2 levels in RTT patient-derived fibroblast cell lines in a dose-dependent manner. Importantly, the increases are capped at non-toxic levels due to the modest impact of each miRNA on gene regulation. Ongoing studies aim to assess the efficacy of sbASOs in RTT-derived iPSCs neurons and in vivo, to explore whether the elevated expression of mutant MeCP2 variants is associated with improved function and mitigation of pathological features.

The Association Between Neighborhood Socioeconomic Deprivation and Colorectal Cancer Screening: Does Modality Matter?

Lauren Giurini ‘201 , Wei Zheng MD PhD2 , Loren Lipworth PhD2 , Harvey J Murff MD3 , Shaneda Warren Andersen PhD1,2,4

1 Department of Population Health Sciences, School of Medicine and Public Health, University of Wisconsin-Madison, 610 Walnut St, WARF Office Building, Suite 1007B, Madison, WI 53726, USA
2 Division of Epidemiology, Department of Medicine, Vanderbilt Epidemiology Center, Vanderbilt-Ingram Cancer Center, Vanderbilt University School of Medicine, 2525 West End Avenue, 8th floor, Suite 800, Nashville, TN 37203-1738, USA
3 Department of Medicine, Vanderbilt University Medical Center, 6012 Medical Center East, 1215 21st Avenue South, Nashville TN, 37232, USA
4 University of Wisconsin Carbone Cancer Center, Madison, WI, 53726, USA

Racial and socioeconomic disparities in colorectal cancer (CRC) outcomes persist, despite effective, preventive screening modalities. Such modalities include colonoscopy and sigmoidoscopy, which require an in-person doctor’s visit, bowel preparation, and an invasive test. Additionally, effective stool-based screening modalities can be done at home, require less prep, and are considered much less invasive. We hypothesized that stool-based tests may alleviate barriers to screening that may be a result of low neighborhood SES, such as transportation, and distance to a provider, and may be taken up by participants in low-income neighborhoods at a higher rate. Of the 85,000 participants of the Southern Community Cohort Study (SCCS), we utilized data from 47,740 who were eligible for screening at baseline, by guidelines from the American Cancer Society and US Preventive Services Task Force, and 30,191 who were eligible at follow-up. The SCCS is a large, longitudinal cohort of English-speaking adults aged 40-79 residing in the southeastern United States and is comprised mostly of participants self-identifying as Black and low-income. Neighborhood-level SES is measured via a Neighborhood Deprivation Index constructed from census-tract variables in the domains of housing, education, employment, occupation, and poverty. Participants self-reported screening with colonoscopy, and sigmoidoscopy at the baseline interview 2002- 2009 and additionally reported stool-based testing at the first follow-up in 2008-2012. Multinomial logistic regression models were utilized to assess the association between neighborhood-level SES and testing by each screening modality. Overall, 43% of those eligible at baseline had been screened prior to baseline, and 59% of those eligible at follow-up had been screened between the baseline and the follow-up interview. Of those eligible for screening at baseline, the percentage of participants reporting no screening increased as neighborhood SES decreased (Q1=42.98%, Q2=49.58%, Q3=52.53%, Q4=56.39%, Q5=62.86%), but of those that did report screening, colonoscopy was more commonly used over sigmoidoscopy (Q1=31.71%, Q2=29.47%, Q3=28.82%, Q4=25.77%, Q5=20.45%). Of those eligible at follow-up, the percentage of participants reporting no screening also increased as neighborhood SES decreased; however, stool-based screening was most common in every quintile of neighborhood SES (Q1=46.67%, Q2=41.01%, Q3=40.69%, Q4=43.0%, Q5=41.16%). The majority of odds ratios calculated at both time points indicated higher odds of reporting no screening over any screening modality and odds ratios comparing screening modalities were null. This study provides evidence that stool-based screening may be a tool to improve the uptake of CRC screening regardless of neighborhood SES and indicates that there may still be barriers to address in order to minimize disparities in CRC screening uptake.

Clinical Outcomes Following Robotic Abdominal Wall Reconstruction For Ventral Hernias Using Resorbable Biosynthetic Mesh

Alexandra C. Skoczek ‘181 , Patrick W. Ruane2 , & Dennis L. Fernandez3

Edward Via College of Osteopathic Medicine – Auburn1, Edward Via College of Osteopathic Medicine – Carolinas2, Crestwood Medical Center3

Background The open approach and the minimally invasive laparoscopic approach have been the mainstay for ventral hernia repair in the United States. While these techniques are successful in repairing ventral hernias, they tend to have high rates of surgical site occurrences (SSO), recurrence rates, and prolonged hospital length of stays. Compared to open and laparoscopic hernia repairs, robotic hernia repairs have been shown to have better postoperative outcomes. However, studies analyzing surgical outcomes following robotic hernia repair are limited in usefulness due to the variety of surgical techniques used. In addition, while previous studies have shown that modifiable comorbidities (MCMs) increase the odds of postoperative complications, no studies have been conducting evaluated MCMs using a robotic technique. Objective The objective of this study is to determine differences in postoperative outcomes for patients with and without MCMs following robotic transversus abdominis release (TAR) with resorbable biosynthetic mesh underlay for primary ventral hernia repair. Methods This study represents a retrospective cohort to assess outcomes for primary ventral hernia repair using robotic TAR with resorbable biosynthetic mesh underlay performed by a single surgeon. Medical records of patients who underwent the robotic hernia repair using this technique between 2015 to 2022 were collected and analyzed for the outcomes of interest for patients with and without comorbid conditions. Three groups were analyzed: patients with 0, 1, and 2+ MCMs. Primary outcomes included surgical site occurrences, prolonged length of hospital stay, and recurrence after 36 months. The study was approved by the VCOM IRB (#2022-079). Results A total of 334 subjects met the inclusion criteria for SSO and prolonged LOS analysis. Of those, 16.8% had no MCM, 56.1% had 1 MCM, and 27% had 2+ MCMs. No significant difference in SSO was seen between the 3 groups, however, having 2+ MCMs was significantly associated with increased odds of SSO (odds ratio 3.25, p=.019). When the groups were broken down, only having a history of diabetes and obesity was associated with significantly increased odds of SSO (odds ratio 3.54, p=.02). Though there was a difference between the three groups, none showed a significantly increased odds of prolonged LOS. Diabetes plus obesity was the only combination of MCMs resulting in increased odds of a prolonged LOS. A total of 175 subjects met the inclusion criteria for recurrence at 36-month follow-up analysis. Of those, 21.1% had no MCM, 48% had 1 MCM, and 30.9% had 2+ MCM. There was no significant difference seen in recurrence rate or significant increase in odds of recurrence between the 3 groups or between the different MCMs and combinations of MCMs. Conclusion The presence of 2+ MCMs significantly increases the odds of SSO, specifically in patients who have a history of diabetes and obesity. However, the presence of any number of comorbidities (0 MCM, 1 MCM, or 2+MCM) was not associated with increased odds of prolonged length of hospital stay or recurrence after 36 months. These findings suggest that the robotic TAR with resorbable biosynthetic mesh underlay for primary ventral hernia repair may be a preferred method of ventral hernia repair in patients with MCM to decrease adverse postoperative outcomes.

Effects of stress on risk assessment behaviors and response to diazepam in the elevated plus maze in adult rats.

Courtney Stickling ’21 (2) and J. Amiel Rosenkranz (1,2)

1 - Discipline of Cellular and Molecular Pharmacology, Department of Foundational Sciences and Humanities, Chicago Medical School, Rosalind Franklin University of Medicine and Science, IL 60064 North Chicago, USA
2 - Center for Neurobiology of Stress Resilience and Psychiatric Disorders, Rosalind Franklin University of Medicine and Science, North Chicago, IL, USA

Anxiety impacts more than a third of Americans at some time during their life. Stress is a major trigger for anxiety. But anxiety has multiple behavioral manifestations and we still do not fully understand what components are sensitive to stress. Risk assessment is an evaluation of the surrounding environment and biased risk assessment can contribute to anxiety. The Elevated Plus Maze (EPM) is a well-established model for assessing and predicting anxiety-related behavior in rodents. The present study uses EPM to investigate risk assessment behaviors in adult rats following a period of repeated stress. We hypothesize that repeated stress in rats will increase risk assessment behaviors and bias decisions after risk assessment towards less risky behavior. This was measured with stretching, rearing, and looking behaviors. We further evaluated the hypothesis by investigating the effects of diazepam, a central nervous system depressant and anxiolytic, on risk assessment. We propose that diazepam will reduce instances of risk avoidance and increase risk taking behavior after risk assessments. Repeated social defeat stress (RSDS) was modeled to expose adult rats to repeated stress. We found that stress increases some risk assessments and sequential behaviors after making the assessment. Open arm stretching was longer in stressed rats compared to non-stressed rats, and stressed rats in general spent less time in the open arm. Furthermore, stress increased the likelihood of risk assessment stretch behaviors prior to entering an open arm, and decreased the likelihood that a rat would enter an open arm after risk assessment. While rearing and looking behavior were not sensitive to stress in this assay, they are consistent with exploratory behavior that occurs after risk decision making took place. Diazepam was also found to ameliorate the effects of repeated stress within the EPM for both risk assessment and exploratory behavior. These results provide insight into which behaviors indicate risk assessment and what type of decisions are made after risk was evaluated.

Diabetes and colorectal cancer in traditionally underrepresented research participants: A prospective cohort study

Zoe L Walts ‘211,2*, Thomas Lawler, PhD1*, Timothy Su, MD, PhD3 , Qiuyin Cai, MD, PhD3 , Mark Steinwandel4 , Wei Zheng, MD, PhD, MPH3 , Shaneda Warren Andersen, PhD1,2,3
* These authors contributed equally to this work

1University of Wisconsin Carbone Cancer Center, Madison, WI, 53726, USA
2Department of Population Health Sciences, School of Medicine and Public Health, University of Wisconsin-Madison, 610 Walnut St, WARF Office Building, Madison, WI 53726, USA
3Division of Epidemiology, Department of Medicine, Vanderbilt Epidemiology Center, VanderbiltIngram Cancer Center, Vanderbilt University School of Medicine, 2525 West End Avenue, 8th floor, Suite 800, Nashville, TN 37203-1738, USA
4 International Epidemiology Field Station, Vanderbilt Institute for Clinical and Translational Research,1455 Research Blvd., Suite 550, Rockville, MD 20850, USA

Background: Type 2 diabetes and colorectal cancer (CRC) disproportionately burdens Americans of lower socioeconomic status and Black racial identity. Despite established population-level disparities, the welldocumented association between diabetes and CRC has been understudied in these demographic groups. We evaluate the association between diabetes and CRC risk in a cohort primarily composed of low SES and Black participants. Methods: Data arose from 74,827 participants of the Southern Community Cohort Study. At enrollment, most participants self-reported Black identity (68%), or annual household income <$15,000 (56%). Diabetes and diagnosis age were self-reported at enrollment (2002-2009) and three subsequent follow-ups. Incident CRC after study enrollment (n=901) was assessed via linkage to state cancer registries and the National Death Index. Hazard ratios (HRs) with 95% confidence intervals (CIs) were obtained via Cox proportional hazards models, using age as the time scale and a time-varying diabetes exposure. Results were stratified by several possible confounders. Results: Diabetes diagnosis was associated with higher CRC risk (HR [95% CI]: 1.35 [1.13-1.62]), with consistent results across tumor location, race, sex, and screening history. Increased risk was observed for current (1.50 [1.07-2.09]) and former (1.93 [1.35-2.76]), but not never smokers (1.00 [0.76-1.32]). This positive association was strongest among participants diagnosed with diabetes within five years of incident CRC (1.64 [1.27-2.14]). Conclusions: Diabetes was associated with elevated CRC risk in this cohort representing an understudied population subgroup. Analyses of majority White cohorts have found similar associations. The strength of the association of CRC with recent-onset diabetes suggests a critical exposure window related to CRC risk. In Black, low SES populations, diabetes prevention or control may reduce cancer burden, particularly for those with a history of smoking.

Current Student Abstracts

Quantification of Collagen in Muscle Tissue of a Longitudinal Murine Model of Cancer Cachexia

Anne Ahern ‘23, Amber Willbanks, Ishan Roy, Biologics Lab, Shirley Ryan Ability Lab, Chicago, IL, 60605

Cancer cachexia is a debilitating, multi-factorial syndrome characterized by involuntary weight loss, muscle wasting, and systemic inflammation. Understanding molecular changes in essential for elucidating the mechanisms of muscle wasting in cancer cachexia. The structural protein collagen is a molecule found in muscle that contributes to the elasticity, force transmission, and contractile strength of muscle fibers. Increases of collagen in human skeletal muscle have been consistently reported in patients diagnosed with pancreatic cancer cachexia. Cancer cachexia has a high incidence in pancreatic cancer patients, with greater likelihood of presentation as a disease progresses. Therefore, a longitudinal mouse model using interperitoneally injected pancreatic ductal adenocarcinoma was appropriate. We hypothesize that the skeletal muscle tissue of mice later in the disease progression of pancreatic ductal adenocarcinoma would have an elevated level of relative collagen presence compared to earlier timepoints and controls. We biochemically analyzed mouse tibialis anterior muscle tissue specimens obtained from age-matched control and pancreatic cancer dosed mice using a hydroxyproline assay. By quantifying the hydroxylated form of the amino acid proline using UV-Vis spectroscopy and a colorimetric assay, we calculated the amount of collagen present in the tissue samples and relate it to the functional ability and cancer status of the model mice. Using a two-tailed student t test, our results indicate that animals relatively far along in disease progression (week 7) had a significant increase (p= 0.0047) in collagen content per mg of muscle tissue compared to controls. Earlier timepoint (week 6) mice were not found to have a significant difference (p= 0.3947) compared to age-matched controls indicating a potential role for collagen in the pathogenesis of muscle wasting associated with cancer cachexia on a temporal basis. This descriptive knowledge has potential to illuminate the mechanisms through which collagen increase may affect muscle function in the later progression of cancer cachexia. We recommend further research on the effects of an increase in relative presence of collagen in cachectic muscle on muscle mechanics to better contextualize our results and their application, and as a result, inform development of new therapeutic strategies.

The impact of pregnancy on mood, cognition, and microglia function in Alzheimer's disease mice

Perla Arias ‘25 and Dr. Holly Hunsberger, Center for Neurodegenerative Disease and Therapeutics, Rosalind Franklin University of Medicine and Science, North Chicago, IL, 60064

Alzheimer’s disease (AD) impacts six-million Americans, 2/3rds of whom are women. Women are more susceptible to developing AD due to a variety of social stress factors, differences in immune responses, and a decline in estrogen. Specifically, Fox et al. (2018) notes that a woman's cumulative months pregnant is indicative of a lower AD risk. Notably, pregnancy has the ability to modify a woman’s inflammatory response. This interaction between pregnancy and inflammation is critical given that there is microglioglial upregulation to clear amyloid plaque burden in the brain. In some instances, the activation of microglia can promote the phagocytosis and clearance of those toxic proteins, but if microglia become overactivated, a surplus of inflammatory mediators are released. Here we aim to understand how pregnancy impacts cognition and microglia activation in Alzheimer’s disease mice. To answer these questions, we used behavior testing and immunohistochemistry to analyze cellular activity. We found that 1) litter-bearing mice exhibit slight memory impairment with age and 2) litter size was indicative of memory impairment in AD mice. Furthermore, immunohistochemistry using microglia markers will give insights into the complexity of microglia function in naive and litter-bearing control or AD mice. With the addition of neurogenesis staining, this research will also provide insight into the detrimental pathology that follows amyloid plaque accumulation. Overall, this research provides a deeper understanding of how pregnancy influences brain pathology and AD symptoms.

A biochemical approach to Mtb; Investigating novel drug targets within the pathogen

Alvaro Arroyo ‘26 and Dr. William Conrad, Biochemistry & Molecular Biology Department, Lake Forest College, Lake Forest IL 60045

Mtb is a merciless pathogen with increasing occurrence of antibiotic resistance and long treatments; new drugs are needed. Once inside a host, Mtb will engage in a fundamental battle between the host’s macrophages. As a means of putting an end to this invasion, the host’s immune cells will surround and enclose the Mtb forming what is known as a granuloma: a trademark of this infection. Despite this effort by the host, the bacterium have the ability to withstand and propagate within said granulomas. Mtb contain transmembrane protein Rv2707, which our lab discovered for survival within intracellular and extracellular compartments of the host. Previous research has demonstrated that in the absence of Rv2707, M. tuberculosis is attenuated in vivo, but not in vitro. This has been demonstrated by research on MMAR_2006, an ortholog of Rv2707 (Will Jiang Cambridge 2018). The precise molecular mechanisms of Rv2707 remain unknown. To further knowledge on Rv2707, we set out to express Rv2707 within E. coli cultures and purify via his-tag purification. On top of expression and purification, we focused on refining our purification methods with hopes of expanding our purified protein yield. We discovered that yield is maximized when e. coli are grown under 16°C, the vector is induced by IPTG, and enzymes are used for cell lysis. The future outlook is to mass produce Rv2707 and incorporate it into giant unilamellar vesicles to conduct functional tests on properties such as ion channel activity. Overall, this research aims to expand current understandings of how Rv2707 protects Mtb against the host, so we can inhibit it and break the malignant chain of transmission.

Finding the right chemical agent to ablate the lateral line system of silver hatchetfish (Gasteropelecus sternicula) for future behavioral experiments

Anais Azevedo ‘26, Kyle Lassen ‘25, Jeremy Levin ’26, Jia Zheng ’25, and Margot Schwalbe, Biology Department, Lake Forest College, Lake Forest, IL 60045 

The lateral line system, a hair-cell based sensory system unique to fish and some aquatic amphibians, plays a pivotal role in several behaviors, including navigation, by detecting water flow and pressure changes immediately around the body. This specialized sensory organ is critical for a fish’s ability to perceive its hydrodynamic environment, aiding in crucial behavioral functions such as prey detection, predator avoidance, and bodily orientation. The lateral line system comprises a network of canal and superficial neuromasts distributed on the head and along the body and tail. Here, we studied the silver hatchetfish (Gasteropelecus sternicula), which have two very unique morphological features: a large pectoral girdle that allows them to jump out of the water and a very prominent and distinctive lateral line system. The goal of this study was to investigate the efficacy of gentamicin and neomycin, two aminoglycoside antibiotics, and cobalt chloride, a heavy metal, in disabling the lateral line system of silver hatchetfish. Adult silver hatchetfish were exposed to varying concentrations of these chemical agents and for specific time intervals. Fluorescence imaging and quantification of lateral line neuromasts were performed following the chemical treatments to assess the treatment's effects on the lateral line system. Our results demonstrate that gentamicin treatment effectively ablates the lateral line system in a dose- and time-dependent manner. The fluorescence imaging confirmed the absence of lateral line neuromasts in treated individuals, supporting the efficacy of gentamicin in disrupting this sensory organ. This study lays the groundwork for future behavioral experiments investigating the role of the lateral line system in the jumping behavior of silver hatchetfish, providing a potential avenue for further understanding of the mechanisms underlying their unique locomotor abilities.

Alprazolam Impairs Fear Memory and Alters Dorsoventral CA1 Neuronal Ensembles in Female Mice

Naina Beishembieva ’25, K. Kaplan, L. Toennies , H. C. Hunsberger, Center for Neurodegenerative Diseases & Therapeutics, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Benzodiazepines are a class of anxiolytic drugs prescribed to patients to treat anxiety, insomnia, seizures, panic disorders, and psychiatric conditions. However, benzodiazepines are also known to interrupt new memory formation along with other side effects. Although benzodiazepines are commonly used and prescribed, the brain regions and neuronal mechanisms responsible for this detrimental side effect are unknown. The purpose of this study was to determine if the benzodiazepine Alprazolam could impair the ability to acquire new memories and whether neuronal activation was altered. We used a contextual fear learning task to measure memory impairment and our activity dependent tagging mouse line (ArcCreERT2 x eYFP) to quantify cells active during encoding and retrieval of the context. ArcCreERT2 x eYFP were injected with Alprazolam or saline 30 minutes prior to a 3-shock contextual fear conditioning procedure. 5 days later mice were placed back in the context without shock and freezing behavior (a proxy for memory) was assessed. 1 hour after testing, mice were euthanized and brain tissue was analyzed for EYFP (encoding cells), c-fos (retrieval cells), and overlap cells (memory traces). We found that female mice showed impaired contextual fear memory during re-exposure (less freezing). In male mice, we found no significant memory deficit during contextual fear learning, however there was a reduction of EYFP indicating that Alprazolam still had an effect on the cellular level, which could lead to long-term changes in memory. Overall, our results suggest that Alprazolam can impair the ability to acquire new memories. Because this is a static view of memory mechanisms, we can next utilize our miniscope calcium imaging system to view cells firing in real time while the animal is injected with drug or during a behavior test.

Insight into Synucleinopathies: Molecular Dissection of Beta- and Gamma-Synuclein for potential toxicity in a yeast model

Federica Bertolotti ‘24, Holly Kiernan ‘26, Tracey Nassuna ‘23, Ryan Osselborn ‘23, Leslie Casares ‘26, and Shubhik DebBurman, Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

The family of synucleinopathies are linked to the misfolding and aggregation of proteins within the family of the synucleins (α-, β-, and γ-), include Parkinson’s Disease (PD), the second most prevalent neurodegenerative disease. While α-synuclein is well-studied for its direct contribution to PD, less is known about the role in neurodegeneration and toxicity potential of β- and γ-synuclein. Two β-synuclein mutants (P123H and V70M) were recently linked with Dementia with Lewy Bodies (DLB), and recently, γ-synuclein inclusions were reported observed with ALS pathology. Lessons from α-synuclein pathogenicity demonstrate that it is not only enhanced by point mutations within it, but modified by post-translational modifications and altered cellular environments linked with mitochondrial dysfunction, altered lysosomal pathways, nitrative stress, and lipid metabolism. Here, we further evaluated the toxicity potential of β- and γ-synuclein in these various neurodegeneration-related cellular environment strains, using our Saccharomyces cerevisiae (budding yeast) PD model system. Additionally, we evaluated substitution mutants for V70M and P123H β-synuclein, where the original amino acid was mutated to representatives of all four amino acid classes (A, R, N, E), for whether loss of the original amino acid (V70, P123) or gain of the new mutant (70M, 123H) is key to toxicity. Finally, we swapped several known familial mutations in α-synuclein and β-synuclein onto each other and onto γ-synuclein as assessed their toxicities. We report that: 1) V70M and P123H β-synuclein mutants aggregate and are more toxic than WT β-synuclein, in a strain- and expression-dependent manner. 2) Evaluation of β-synuclein substitution mutants demonstrate that that gain of histidine is key to P123H-β-synuclein toxicity. 3) The V70M b-synuclein mutation when swapped into a-synuclein makes the latter differentially more toxic than swapping in the P123H mutation. 4) WT and mutant β-synuclein toxicities are differentially aggravated by altered nitrative stress. Overall, this study expands the evaluation of β- and γ-synuclein, as well as genetically modified mutants in a yeast model, to understand mechanisms of toxicity for these two nervous system proteins and illuminates mutant toxicity in β-synuclein.

Determining Phylogenetic Trees Using Algebraic Statistics

Emily Bross ’25 and Sara Jamshidi, Data Science Department, Lake Forest College, Lake Forest, IL 60045

Inferring phylogenetic trees—the process of matching an evolutionary tree to existing species— is a fundamental challenge in evolutionary studies. While there are mathematical processes to construct trees using clustering methods, this project tests and verifies an information-theoretic approach from Algebraic Statistics that matches DNA data to a proposed phylogenetic tree. The methodology uses vectors to capture the relative frequency of base pairs from a sufficiently informative DNA segment of a set of existent species. A tensor is constructed from these vectors based on the order of the leaves within the phylogenetic tree. To capture the algebraic geometry of the proposed tree, the tensor is tested by assessing tensor flattenings based on internal edges of the tree, after the phylogenetic tree is heavily simplified. While this methodology is sound from an information theoretic perspective, not much work has been done to demonstrate the efficacy using real DNA data. Here, we write a Python script from scratch to replicate this process with both synthetic and ribosomal DNA data. In the case of real data, we compare the DNA to a tree generated by Clustal Omega and verify the tree does match the DNA with this technique—a fundamentally different approach from the mathematics used to construct the tree (“Clustal Omega”).

The effects of pollen limitation on life history fitness in long-lived perennial coneflower, Echinacea angustifolia

Sophia Chen ’23 and Stuart Wagenius, Department of Plant Conservation Sciences, Chicago Botanic Garden, Glencoe, IL 60022

Pollen limitation is an event where plant populations have limited access to pollen. This effect has been observed in remnant plant populations and leads to negative consequences on reproductive fitness. However, few studies have provided a comprehensive analysis on the effects of life history fitness in perennial plants. This study aims to investigate pollen limitation effects on components of life history fitness through the Aster Model Analysis. This mathematical model takes a holistic approach to assessing life history fitness in perennial plant species by taking into account allocation to survival and reproduction at multiple time points throughout an individual’s life cycle. Echinacea angustifolia is used as a model perennial plant as it is self-incompatible and past studies found them to be pollen-limited in fragmented prairie remnants. A cohort of common garden E. angustifolia has been separated into a pollen exclusion group and a pollen addition group to measure life history fitness over the course of 12 years. We hypothesize that pollen limitation may elicit a trade-off in reproductive fitness by alternate flowering patterns, number of heads produced and decrease mortality.

Prey Detection: Exploring vision and the lateral line system of Bumblebee Goby Brachygobius doraie

Shrija Chhetri ‘24, Djina Jovanovic '23, Violet Anderson '23, and Margot Schwalbe, Biology Department, Lake Forest College, Lake Forest, IL 60045 

The mechanosensory lateral line system of fishes is thought to form hydrodynamic images of flow disturbances immediately around the body and can mediate several behaviors, including prey detection. The lateral line system is composed of a stereotypical array of neuromasts on the body, which can be located within bony canals (canal neuromasts), and on the skin (superficial neuromasts). There are four types of lateral line canal patterns among teleost fishes and one type is called reduced cranial canals. This canal type is characterized by a reduction of canals on the head, the absence of a trunk canal, and extensive proliferation of superficial neuromasts on the body. Further, taxa with proliferation of superficial neuromasts tend to occupy hydrodynamically quiet and/or light-limited environments, and more superficial neuromasts on the skin may increase the sensitivity to local flows and to prey. A diverse group of fishes called gobies (~1360 species) have reduced canals, including the bumblebee goby (Brachygobius doraie). Few studies have linked this lateral line canal pattern to prey detection, and thus we asked the question – how does a goby use its reduced lateral line canal system to find prey? We tested the role of vision and the reduced lateral line canal system of the bumblebee gobies during feeding trials with live, mobile prey under light and dark conditions, and with an intact or chemically ablated lateral line system. We evaluated several parameters of search and prey detection behaviors from recorded feeding trials, including number of prey eaten, prey detection and reaction distances and angles, and time to first strike. Through our experiment, we demonstrated that bumblebee gobies can localize and capture prey under both light and dark conditions, and they perform different search strategies depending on available sensory systems. Prey detection behavior involves a combination of several sensory modalities; depriving a goby of a single sensory system impacts its ability to detect prey. This study provides important insights into the functional and ecological significance of lateral line-mediated behavior in a species with reduced lateral line canals.

Precision mapping of neuronal activity in sea slug motor programs reveals spatial organization of functional clusters

Elma Choi ‘25, Viral Mistry, Jeffrey Brown, Evan Hill, and William Frost, Stanson Toshok Center for Brain Function and Repair, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Neurons organize themselves into networks in order to produce complex behaviors. By recording their activity during such behavior, it is possible to identify clusters of cells with similar firing patterns and thus potentially similar functions. However, the ways in which these functional clusters relate to the actual spatial organization of neurons in the brain remain elusive. Using isolated Tritonia diomedea and Aplysia californica brain preparations stained with voltage-sensitive dye, we optically imaged the activity of neurons in the pedal ganglia during escape locomotion motor programs. We then used computer scripts to map neural traces back onto their cells of origin on a photograph of the pedal ganglia, create functional clusters based on similarity between traces, and color-code the mapped cells according to which cluster they belonged to. We found that cells belonging to the same functional cluster had a tendency to also be discretely spatially clustered, consistent with a spatial-functional organization of different sub-groups within the locomotion motor programs. Future work will identify how learning changes these subgroups and if they represent dedicated re-identifiable neuronal sub-populations within the motor program.

The Cx50D51A mutation displays increased hemichannel activity.

Kesang Y. Dorji ’25 and Lisa Ebihara, Department of Biophysics and Physiology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Undocked connexin hemichannels are found in the plasma membrane of most cells. Under physiological conditions, they are maintained mainly in the closed state by the binding of calcium to the extracellular side of the pore. This property appears to be critical for the maintenance of normal cellular homeostasis. Connexin mutations that result in increased hemichannel activity have been associated with decreased cell survival and are implicated in a number of inherited diseases including KIDS syndrome and congenital cataract. However, the molecular mechanisms of calcium-induced hemichannel closure remain unclear. Here we examine the role of the conserved charged residue, D51, at the extracellular entrance of the Cx50 hemichannel in extracellular calcium blockade by expressing Cx50D51A in Xenopus oocytes and studying its functional properties using the two-microelectrode-voltage-clamp technique. Our results show that alanine substitution at position 51 resulted in greatly enhanced hemichannel currents with altered voltage-gating properties compared to wild-type Cx50. These enhanced currents were associated with a reduction in cell resting potential and increased cell death. To determine whether the D51A channels were inhibited by external calcium, currents were recorded for cells in 0, .2, 1, 3 mM calcium containing MB solutions. Increasing external calcium reduced but did not completely block the hemichannel currents. Co-expression of mutant D51A with wild-type Cx50 resulted in hemichannel currents whose properties were indistinguishable from those induced by D51A alone, suggesting that the mutant had a dominant effect. These results suggest D51A stabilizes the Cx50 hemichannels in the open state and decreases cell survival but does not completely eliminate the effect of external calcium.

Insight into Synucleinopathies: Comparative Evaluation of Wild-Type Alpha, Beta-, and Gamma-Synucleins in a Yeast Model

Sebastian Gacek ‘25, Leslie Casares, Federica Bertolotti ‘24, Isaiah Moonlight ‘25, Ryan Osselborn ’23, Marcelo Camba Alamban ‘26, and Shubhik DebBurman, Neuroscience Program, Lake Forest College, Lake Forest, IL 60045

Synucleinopathies are a group of neurodegenerative disorders linked with the misfolding and aggregation of ⍺-synuclein, the most well-known among them being Parkinson’s disease (PD). ⍺-Synuclein belongs to a larger family of proteins that include β- and 𝛾-synuclein. Mutant forms (P123H and V70M) of β-synuclein have been shown to cause Dementia with Lewy Bodies (DLB). However, the extent to which β- and 𝛾-synuclein are neurotoxic is still highly understudied compared to ⍺-synuclein. While specific alterations in cellular environments (including nitrative stress, lysosomal degradation, mitochondrial dysfunction) and post-translational modifications alter the toxicity and aggregation of α-synuclein, less is known of their impacts on β- and 𝛾-synuclein. Here, we used our lab’s budding yeast (Saccharomyces cerevisiae) model to comparatively evaluate these three wildtype synucleins and explore their pathological potential through the assessment of toxicity, localization, and expression. We report that: 1) Both wildtype ⍺- and β-synuclein are differentially toxic in control yeast strains, whereas 𝛾-synuclein is non-toxic. 2) Expression levels of these proteins are tied to extent of aggregation and cytotoxicity observed. 3) Toxicity potential of all synucleins are impacted to varying degrees by specific synucleinopathy-related altered cellular environments in genetically altered yeast strains. 4) β-synuclein, when expressed in budding yeast, displays a higher molecular weight than expected. This study adds to a growing body of research exploring disease association of the synuclein family of proteins and illustrates the importance of further evaluation of β-synuclein’s role in neurodegeneration.

Identification and Analysis of Organic Anion Transporter 1 (OAT1) Through Western Blotting and Fluorescent Uptake Assays – Development and Testing of Anti-HIV Agents.

Jorge A. Garcia ‘24, Natalia Rak, Yalitza Jimenez, and John K. Buolamwini, Department of Pharmaceutical Sciences, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Human immunodeficiency virus (HIV) is a viral infection that causes acquired immunodeficiency syndryme (AIDS). It infects and weakens the host’s immune system by killing white blood cells, specifically CD4 T lymphocytes (Doitsh & Greene, 2016). If HIV infection is not treated, then it proceeds to develop into acquired immunodeficiency syndrome (AIDS) (HIV and AIDS, n.d.). Tenofovir is a common antiretroviral therapy that effectively targets HIV (Rodríguez-Nóvoa et al., 2009). Although Tenofovir is a great anti-HIV agent, it has also been linked to cause severe kidney tubular dysfunction (KTD) which can be lethal (Rodríguez-Nóvoa et al., 2009). Tenofovir can enter epithelial cells of the kidney tubule by way of organic anion transporters (OATs) – specifically OAT1 – since they are expressed in the basolateral membrane of kidney cells (Rodríguez- Nóvoa et al., 2009). Because Tenofovir can cause lethal damage to its host, it is imperative that new drug treatments and therapies are developed that are effective and minimize side effects. OATs are ideal for studying substrate binding because they can also bind to carboxylfluoresceins (CFs) – fluorescent compounds that bind to OATs with higher affinity and are transported by them – and thus, measure fluorescent uptake through fluorescent-based assays (Truong et al., 2008). In this experiment, human kidney 2 (HK-2) cells were used since they retain functional characteristics of proximal tubular epithelium cells (Ryan et al., 1994). HK-2 cells therefore should have the OAT1 protein and thus, we can use CFs along with multiple substrate inhibitors and pro-drugs – including Tenofovir – to study OAT1 fluorescent substrate uptake. 6-carboxylfluorescein (6-FAM) was used as the substrate in this experiment since it has a stronger affinity for OAT1, the protein of interest. Furthermore, it was also essential to demonstrate the expression of the OAT1 protein on the HK-2 cells, so a protocol was developed to isolate and perform Western blotting to detect OAT1 protein. The OAT of interest does appear on the western blot development and there is differential fluorescent substrate uptake with varying inhibitors and pro- drugs – including an inhibitor that is just as effective as Tenofovir. Thus, this new inhibitor is of interest and will be foundational for future experiments.

AI-Based Characterization of 6-OHDA-Induced Parkinsonian Motor Symptoms in Mice

Ying Han ’24, Department of Neuroscience and Data Science, Lake Forest College, Lake Forest, IL 60045, Terry Cirrencione, Nadine Pickering, Sayli Korde, Eun Jung Hwang, Discipline of Cell Biology and Anatomy, Stanson Toshok Center for Brain Function and Repair, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, 60064

Parkinson’s disease (PD) is a prevalent neurodegenerative condition that is associated with progressive loss of dopaminergic neurons. Prominent PD motor symptoms include tremor, slowness, rigidity, and postural instability. Animal models of PD are invaluable for understanding the disease mechanisms and developing effective treatments. Behavioral assessments are essential for evaluating the efficacy of potential PD treatment options in these animal models. To enhance the precision, reproducibility and efficiency of behavioral assay in animal PD models, we set out to leverage advanced artificial intelligence (AI) technology to detect and score PD symptoms. Toward this goal, we first generated PD mice through intracranial injection of neurotoxin 6-OHDA into the right striatum, which selectively degenerates dopaminergic neurons. During the interval of 1 day before and 21 days after the surgery, we video-recorded free-roaming behavior of these mice and extracted movement data from their key body parts using an AI machine vision tool, DeepLabCut. Then, we characterized the changes in various kinematic metrics including the total distance traveled, roaming, speed, freezing duration, ipsilateral turnings, and body length. Validating our 6-OHDA model, mice became hypokinetic, i.e., decreased travel distance and speed and in total travel distance. Furthermore, PD mice exhibited an increased propensity for rightward turns as expected from the unilateral brain lesion on the right hemisphere. Immunohistochemistry further confirmed the unilateral degeneration of dopaminergic neurons. No difference in body length was observed. Statistically significant changes in kinematic metrics were not always readily discernible through visual inspection. This suggests that machine learning algorithms utilizing kinematic data as input have the potential to outperform human observers in the detection and quantification of PD symptoms. Our future work aims to utilize kinematics data to train machine learning algorithms to not only discriminate between PD mice and non-PD mice, but also to compute grades symptom severity scores. Such algorithms will enhance our understanding of PD in animal models and pave the way for more precise evaluations and potential therapeutic interventions.

The Influence of Flooring Surface on Walking Mechanics of Persons with Diabetes

Salman Javed ‘24, Sanem Jhaveri, Brittany Cook, and Ryan Crews, Center for Lower Extremity Ambulatory Research (CLEAR), Rosalind Franklin School of Medicine and Science, North Chicago, IL 60045

Diabetic foot ulcers (DFUs) are critical wounds that form on patients with diabetes. Particularly, the loss of sensation on the feet caused by diabetic peripheral neuropathy leads to the formation of DFUs. Therapeutic shoes are used to protect the feet of persons with diabetes from excessive loading. However, adherence is an issue, particularly in the home. This study examined the effect of low pile, high pile, and non-carpeted tile on the pressure between foot and surface (peak plantar pressure) during walking and standing trial with shoes (shod) and barefoot. Insoles designed to measure plantar pressure were configured to collect 9 plantar masks covering all regions of the feet. Kinematic sensors were used to ensure walking speed did not differ between trials. Trials were conducted to confirm that walking barefoot on all flooring surfaces resulted in higher peak pressure than in shod walking on participants at risk for diabetes or exhibiting peripheral neuropathy.

Identifying Genes That Regulate CaV2/UNC-2 Channel Levels at the Presynaptic Terminal in C. elegans

Wambui Kahende ’24 and Hongkyun Kim, Department of Neuroscience, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

The C. elegans CaV2/UNC-2 Channel is a voltage-gated calcium channel that functions at the presynaptic terminal. We study the mechanisms that regulate UNC-2 levels at the pre-synaptic terminal. Our interest is in discovering novel genes regulate UNC-2 channel levels, and finding the location of these genes in the C. elegans genome. To identify the location of the genes, we use genetic mapping based on single nucleotide polymorphism between two different strains, Bristol N2 and Hawaiian CB4856. Because these genes are expected to be conserved with human homologous genes, the discovery of these novel genes will reveal the possible causes of human diseases such as ataxia and seizure which are caused by calcium channels.

Measuring Spasticity of Lower limbs in Humans with Spinal Cord Injury

Hridey Kapoor ‘251,2, Dalia De Santis2, and Monica A. Perez2,3,4, 1Department of Neuroscience, Lake Forest College, Lake Forest IL, 60045, 2Shirley Ryan Ability Lab, Chicago IL, 3Department of Physical Medicine and Rehabilitation, Northwestern University, Chicago, IL, 4Edward Hines Jr., VA Hospital, Hine, IL, USA.

Lower Limb Spasticity, especially in the quadriceps, is a common symptom after damage to upper motor neurons (UMN). Approximately 70-80% of individuals with Spinal Cord Injury (SCI) develop spasticity in the early stages of their recovery, which may lead to muscle weakness, pain and decreased mobility. Spasticity is a lifelong symptom and there is a need to quantify its severity for tailoring pharmacological treatments and therapeutic interventions. In the clinical settings, spasticity is subjectively rated though scales like the Modified-Ashworth Scale (MAS), which is simple to administer but has poor inter-rater reliability and validity especially in the lower-limb. Quantitative tests mostly used in the research setting, as the Pendulum Test, are a better alternative to provide sensitive and repeatable measurements of quadriceps spasticity that are assessor-independent yet easy to administer.
The pendulum test measures spasticity as the angle of maximum flexion of the leg about the knee during gravity-induced swings. The Pendulum test can be instrumented in many ways, but here we highlight two. The first method is commonly used in research settings (MoCap – Utilizing Infrared cameras and emitters) and requires expensive equipment, a controlled environment, calibration time, proprietary software (Motive-OptiTrack), and technical skills. The second method is a new inexpensive and portable system utilizing Inertial Measurement Unit sensors (IMUs – Utilizing a Gyroscope, and an Accelerometer) that is most suitable to use in the clinical setting. We compare the IMU and MoCap systems for accuracy, difficulty in calibrating, the environment required, ease of use, and cost.
The IMU-based system, showed to have consistent results with MoCap while being more cost, time, and resource effective and is a robust and flexible alternative to camera-based motion capture system to quantify knee extensor spasticity following SCI.

Illuminating neural behaviour in C.elegans utilizing optogenetics.

Joysiaeli Kimambo ’25 and Hongkyun Kim, Department of Cell Biology & Anatomy Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Optogenetics is a technique that allows researchers to genetically modify neurons to produce light-sensitive proteins. Chrimson is a light-sensitive protein which is activated by yellow light and increases neuronal electrical activity. Another protein, GtAcR2, responds to blue light and inhibits neuronal firing. This study involves the exploration of optogenetics in the Caenorhabditis elegans model. Stimulation of all C. elegans neurons results in temporary constricted paralysis and a decrease in body length; inhibition of neurons results in flaccid paralysis and an increase in body length. Through genetic manipulations, the varying effect of optogenetics stimulation and inhibition is investigated between two strains of C. elegans, wild type and the GtAcr2::mScarlet::Chrimson transgene worms. Statistical analyses highlight the stimulatory and inhibitory function of yellow and blue light respectively, whereas WT worms exhibit expected behaviour, while the GtAcr2::mScarlet::Chrimson transgene worms exhibit a significant increase in body length when blue light is illuminated, however a non-significant change in body length is observed when yellow light is illuminated. The results indicate that optogenetics provides the most practical and simplest way of increasing or decreasing the activity level of cells so as to evaluate their functional role and the overall C. elegans neuroscience.

Targeting Phospholipase D1 (PLD1) in Kaposi’s sarcoma-associated herpesvirus (KSHV)-associated primary effusion lymphoma (PEL).

Rick Liao ’25, Serdar Gayybov, Khalil Qadri and Neelam Sharma-Walia, Microbiology and Immunology, 3333 N Green Bay Road, North Chicago, IL 60064

Kaposi’s sarcoma-associated herpesvirus (KSHV), which is also known as human herpesvirus 8 (HHV8), is a type of herpesvirus that is linked to Kaposi's sarcoma (KS) and B-cell lymphoproliferative primary effusion lymphoma (PEL). PEL is a highly aggressive cancer that is commonly found in AIDS patients. Unfortunately, PEL patients have a poor prognosis and limited treatment options. Therefore, it is crucial to identify molecular targets of PEL pathogenesis that can be effectively targeted to control malignant cell survival and proliferation.
Lipophosphodiesterase (also referred to as PLD or by its superfamily phospholipase) is an enzyme found in mammalian cells, and two genes encoding PLD1 and PLD2 have been sequenced and manipulated. Previous research has shown that PLD regulates the actin cytoskeleton and that actinin and tubulins are necessary structural proteins for cytokinesis. The stabilization or disruption of these proteins can affect cell movement and division. Our lab has conducted research that showed PLD1 inhibitor (VUO155069) promotes apoptosis in PEL cells. We demonstrated that PLD1 inhibitor treatment blocked the metabolic pathway regulator mammalian target of rapamycin (mTOR) signaling. Here, we hypothesized that targeting PLD1 would affect the microtubule organization, cell survival, and proliferation of PEL cells, ultimately leading to apoptosis.
As part of this study, we treated the KSHV-infected BCBL-1 and BC-3 and KSHV uninfected BJAB PEL cell lines with a PLD1 inhibitor. We then prepared whole cell lysates and tested for total tubulin content using Western blotting and specific antibodies. Our results showed no change in the tubulin content in PEL cells following PLD1 inhibitor treatment. Studies are currently underway to determine the mechanism of PLD1 inhibition-induced cell death in PEL cells.

Impact of 4-1BB on the Germinal Centers during Plasmoidum infection

Taylor Lyons ’24, Carolina Caloba, Kateryna Malkina, and Rahul Vijay, Centre for Cancer, Cell Biology, Immunology, and Infection, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064

Malaria is a vector borne disease caused by the genus Plasmodium, which are unicellular eukaryotic protozoan parasites with a complex life cycle. In 2021 alone, 247 million cases were reported, many of these in tropical regions1. Although anti-malarial drugs are available to cure an active infection, sterilizing immunity against malaria is not effective. Germinal centers (GC) are not optimally induced following malaria, resulting in weaker immune memory. The purpose of this study was to determine what role 4-1BB plays in the immune response to non-lethal malarial infection through the use of immunofluorescence . 4-1BB is a member of the TNF receptor superfamily and functions as costimulatory molecule that induces T cell activation . By staining for specific immune cell surface proteins in the spleen, once could visualize where different types of cells congregate during Py infection, and whether the absence of 4-1BB impact on this response. Surface markers targeted for staining include B220 (B-cell follicles), GL-7 (GC B cells), CD138 (extrafollicular plasmablasts) and CD4 (T-helper cells). WT and 4-1BB KO B6 mouse spleens were cut at 12 mm thickness, 18 days post Plasmodium infection. The process of creating a successful confocal procedure was trial and error. Initial success came with the addition of Triton X100, which allowed for the permeabilization of cells and the first visual stain of B220. Subsequent success stemmed from staining with a secondary antibody, allowing for visual results with GL-7 and CD4 (CD3e). While CD138 was not successfully stained, results still presented evidence that the absence of 41BB negatively impacted GC immune response, with weaker germinal center formation seen in 4-1BB KO.

Hypoxia Reversal Improves Antimalarial Immune Response

Kateryna Malkina ‘24*#, Lijo John#, Carolina Caloba#, Dr. Rahul Vijay#

*Lake Forest College, Lake Forest, Illinois, 60045
#Center for Cancer, Cell Biology, Immunology, and Infection, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

The effective clearance of the malarial parasite Plasmodium relies on high-affinity antibodies secreted by germinal center (GC)-derived plasma cells (PC) and memory B cells (MBC). However, evidence from malaria endemic regions suggest that naturally acquired anti-malarial immunity from previous malaria episodes are inefficient. Hemolysis and resultant hypoxia are among the main features during malaria, both which have clinical implications. Previously we demonstrated that anemia drives the exposure of phosphatidylserine (PtS) which in turn binds to its receptor AXL on B cells to promote the accumulation of low-affinity plasmablasts (PB) that suppress the GC response during malaria. Whether hypoxia resulting from anemia contributes to this phenotype is still unknown. Here we demonstrate that treating Plasmodium infected mice with an air mixture to reverse hypoxia (95% O2 and 5% CO2) not only reverse hypoxia but also rescues GC response and parasite control. Thus we propose a plausible therapeutic approach to enhance anti-malarial immunity.

TOX1 gene cloning for potential future use to determine its influence on Multiple Sclerosis disease progression in EAE model mice.

Monica Marusarz ’24, Joseph Reynolds, Kristen Patterson, and Allison Coffell, Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Multiple Sclerosis (MS) is an inflammatory auto-immune disease that leads to neurological disabilities caused by the demyelination in the central nervous system. It involves progressive tissue damage mediated by T-cells. Thymocyte selection-associated high mobility group box protein 1 (TOX1) is a gene from the TOX DNA-binding protein family. TOX1 specifically has been found to be crucial in regulating immune system differentiation. TOX1 is also important for the formation of T exhaustion cells which is generally caused by chronic infections. T cell exhaustion is characterized primarily by a loss of cytokine productions as well as impaired proliferation and survival. Here, the TOX1 gene was cloned and sequenced. In the future, the cloned TOX1 gene will be used for in vitro experiments to silence TOX1 genes in mouse spleens and lymph nodes. If successful, in vivo experiments will be done on model EAE mice to determine what effect silencing the gene will have on the disease.

Circadian Control of the Bed Nucleus of the Stria Terminalis (BNST) via Hypothalamic Vasopressin (AVP) Projections

Lorena Monroy ‘241, Susan H. Olson1, Valery Grinevich2, Joanna Dabrowska1, 1Department of Cellular and Molecular Pharmacology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064, 2Laboratory of Neuropeptides, German Cancer Research Center, Central Institute of Mental Health, University of Heidelberg, Heidelberg, Germany.

The bed nucleus of the stria terminalis (BNST) is a limbic forebrain structure that regulates many physiological and behavioral processes, notably, fear and anxiety-like behaviors. The BNST receives inputs from many hypothalamic nuclei that regulate crucial physiological processes (water/electrolyte balance, temperature regulation, feeding, wake/sleep cycle). Our current studies demonstrate that hypothalamic hormone and neuromodulator arginine-vasopressin (AVP) has a direct excitatory effects on neurons in dorsolateral BNST (BNSTDL), particularly neurons expressing striatal-enriched protein tyrosine phosphatase (STEP). However, the source of specific peptidergic inputs containing AVP in the BNSTDL are elusive. Hence, we used AVP-Cre transgenic rats (Cre-recombinase under the AVP promoter) injected with Cre-dependent pAAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby into the suprachiasmatic nucleus (SCN) or the supraoptic nucleus (SON) of the hypothalamus and we show that both these nuclei send AVP-containing inputs to the BNSTDL. To determine whether the SCN and BNSTDL have synchronous activity, we quantified immunoreactivity patterns for AVP and STEP in the SCN and BNSTDL across four zeitgeber times in rats housed in 12:12 light-dark cycles. We used STEP as an indirect marker of BNSTDL neuronal inactivity. We found the highest AVP expression in the SCN occurred at ZT 11, which correspond to the lowest STEP expression in BNSTDL neurons, whereas the lower AVP expression in the SCN and higher STEP expression in BNSTDL occurred at ZT 1. These results suggest that SCN-AVP and BNSTDL-STEP neurons show direct relationship, such as the higher AVP-SCN signals, the more active (expressing less STEP) BNSTDL neurons are. We next used the Period2 Clock protein (PER2), a key gene of the circadian clock, to determine whether the SCN imposes circadian control over the BNSTDL. We found higher PER2 expression at ZT 1 in the BNSTDL and SCN compared to PER2 expression at ZT 11. These results demonstrate an anatomical and functional connection between the SCN and BNSTDL and have implications on how changing physiological conditions, in particular the wake/sleep cycle, through AVP prompt, can directly impact BNST activity and BNST-dependent fear- and anxiety-related behaviors.

Repeated restraint stress induces sex-dependent activation of BNST and anxiety-like before

Rose Jasmin Montes ‘24, Jaime Vantrease, and J. Amiel Rosenkranz, Department of Cellular and Molecular Pharmacology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Anxiety disorders are among the most prevalent psychiatric disorders, with women being twice as likely as men to be diagnosed. This suggests underlying sex differences in the neural circuitry may contribute to the prevalence of anxiety disorders in females. The bed nucleus of the stria terminalis (BNST) is sexually dimorphic, stress-sensitive, and activated during anticipatory anxiety or anticipation of a threat. Prior studies show that female rodents extinguish conditioned freezing to anticipatory anxiety faster than males. However, the extent to which stress modulates anticipatory anxiety in parallel with BNST activation is unknown. Since females are more sensitive to stress, we hypothesized that social interaction would facilitate greater BNST activation in stressed females compared to stressed males and control animals. To test our hypothesis, we used male and female rats exposed social interaction (for 10 minutes) with naïve rats and then to repeated restraint stress (20 min/day for 7 out of 9 days) or control handling. Following treatment, rats underwent another round of social interaction (with naïve rats and for 10 minutes) Afterwards, we immunostained the BNST for c-Fos expression. There was no difference in time spent interacting in the first day of the social interaction. We found significant interactions between sex in that males displayed more avoidance before than females. Preliminary data further suggests that these stress-induced sex differences on anxiety-like behaviors may be related to c-Fos expression in the BNST. We found increased expression in the BNST of stressed rats compared to controls, we also found that they were more prominent in female rats. Together, these studies show how stress influences anxiety-like behaviors in a sex-dependent fashion, possibly through differences in activation of the BNST.

Linguistic Markers of the Dark Triad: A Systematic Review

Amelie Motzer ’24 and Vivian Ta-Johnson, Psychology Department, Lake Forest College, Lake Forest, IL 60045

Although studies have examined the linguistic markers of the dark triad (Narcissism, Machiavellianism, Psychopathy), no systematic review of this body of literature has been conducted to better understand the nature of this relationship. As such, the aim of this present research is to conduct a systematic review of the association between dark triad traits and word use. To ensure coverage, several major databases were searched, and sixteen articles were identified after applying exclusion criteria. Our review revealed a consistent link between Narcissism and the usage of words about Friends, Social Interaction, and self-image.Psychopathy was most consistently linked with the usage of anger words, swear words and words referring to basic needs, particularly money. Machiavellianism was least consistently related to any linguistic markers. Our review also highlights methodological inconsistencies across this body of literature that may impact the field’s understanding of how the dark triad reveals itself through language. As such, we provide recommendations for future research to enable more reliable and robust examinations of the linguistic markers of dark triad traits.

Anatomical Tracing of Convergent Inputs to the Striatum for Decision-making

Ama Owusu-Ofori ‘24, Sayli Korde, and Eun Jung Hwang, Rosalind Franklin University of Medicine and Science, Stanson Toshok Center for Brain Function and Repair, North Chicago, IL, 60045.

Decision-making is a complex process that relies on multiple lines of information including sensory stimulus and internal bias. Previous studies discovered that the posterior parietal cortex (PPC) represents internal bias of individuals and sends that information to the striatum (STR) to influence stimulus-guided decision-making process. However, it remains unknown how the bias information from the PPC interacts with the sensory stimulus-guided decisions. We hypothesize that the STR that receives PPC input also receives input from the primary visual cortex (V1) to integrate the two lines of information. If so, we predict that the axon terminals of cortico-striatal projection neurons from the PPC and V1 are converged in the same region within the STR. To test this hypothesis, we injected viral fluorescence tracers in PPC and V1, each in different colors and inspected their axon terminals in the STR through histological assay. Consistent with our prediction, we found that PPC and V1 project to the same locus in the STR. Based on this result, we plan to examine the causal roles of the V1-STR projection in the activity of STR neurons and decision-making process.

Haptic Feedback Enhanced Mobility (‘White’) Cane Increases Environmental Awareness in the Absence of Vision

Suma Pasupulati ’24 and Frederick R. Prete, Department of Biology, Lake Forest College, Lake Forest, IL 60045

Approximately 23 million people who are blind or visually impaired (B/VI) use mobility canes. Although effective, canes cannot detect obstacles beyond the reach of their tip. We explored the possibility of increasing cane effectiveness by incorporating tip-mounted distance sensors that triggered haptic (vibratory) feedback in the cane’s handle when nearby objects were detected. Experiment 1 compared the accuracy of ultrasonic versus infrared distance sensors by sweeping canes toward and away from a stationary obstacle at four frequencies (40, 50, 60, 70 sweeps/ min). Sensors were most accurate at 40 and 60 sweeps/min which is consistent with cane sweep rates (one sweep/ step) for B/VI users. Optic sensors were more accurate than ultrasonic but were more susceptible to ambient (light) interference. Experiment 2 compared three cane configurations in an obstacle-avoidance task. Canes differed in tip type (roller ball vs stationary), haptic stimuli (short vs long, high vs low frequency), and object sensing range (max=12, 18, 20 inches). Blindfolded volunteers (N=8) tested each configuration by walking back and forth across a room (8 total trips, approx. 40 ft./trip) between two rows of boxes using haptic feedback on alternate trips. We scored the number of box locations detected without touching (identify), prior to touching (bump), or after inadvertently touching (collision) each with the cane. Volunteers identified significantly more boxes and had fewer collisions with haptic feedback. They also preferred canes with roller tips, short duration/low amplitude stimuli, and feedback beginning within 18 inches of potential obstacles.

Investigating the bidirectional relationship between traumatic brain injury and sleep homeostasis in D. melanogaster

Rebecca N. Ray ‘24 and Rebecca Delventhal, Department of Biology, Lake Forest College, Lake Forest IL, 60045

Traumatic brain injuries (TBI) occur when a sudden, severe impact to the head causes brain damage. This can affect short- and long-term physical health, cognition, and homeostatic behaviors, such as sleep. Many mechanisms affected by TBI in humans are also found in Drosophila melanogaster, making them an ideal model to study TBI. Considering sleep is a homeostatic behavior found in nearly all animals, we wanted to investigate the relationship between TBI and sleep and how TBI may affect long-term sleep behavior by using D. melanogaster as a model organism. To test this, we used the High-Impact Trauma (HIT) device to injure wild-type flies and measured their sleep in the days and weeks following TBI. We found that 1week post-TBI, flies had significantly more fragmented sleep than non-injured controls, while 2wks post-TBI, flies slept significantly less than controls. Considering this evidence for TBI-induced disruption in sleep homeostasis, we then asked if disrupting sleep could affect recovery from TBI. We used a genetically short-sleeping fly to test how decreased sleep affects TBI recovery, measured by acute mortality and locomotor function. We found that injured short-sleeping flies had a significantly higher acute mortality than injured control flies with normal sleep. Since there is evidence to suggest a bidirectional relationship between TBI and sleep, we then asked what the underlying molecular mechanism of this relationship might be. Prior research has shown that sleep is related to the clearance of reactive oxygen species (ROS) and the protection against oxidative stress. To investigate whether this association plays a role in the relationship between sleep and TBI, we manipulated the neuronal expression of the antioxidant SOD2, which inversely affects ROS levels, and measured TBI recovery and sleep. Injured flies with either decreased or increased neuronal antioxidant expression showed no significant difference in acute mortality compared to injured controls. Future research manipulating ROS levels outside of the brain could further our understanding of the role of ROS in the relationship between TBI and sleep, which could have implications in new TBI prevention or recovery strategies.

Development of a natural infection model in zebrafish outlines a plausible method for a better understanding of the role of lymphocyte cytosolic protein 1 in tuberculosis infection

Ceylin Sahin ’24, Max Guyott ‘26, Zoe Draper ‘26, and Will Conrad, Biochemistry and Molecular Biology Department, Lake Forest College, Lake Forest, IL 60045

Tuberculosis (TB) remains a global health threat, posing a need for improved research methods to better understand the molecular response to bacterial pathogenesis. TB, caused by Mycobacterium tuberculosis, remains a threat to global health with 10.6 million cases in 2021 alone, reflecting a need for new therapeutic avenues such as host-based therapies. Toward this end, we sought to investigate the role of the host gene lcp1 in tuberculosis pathogenesis. lcp1 encodes the protein product, lymphocyte cytosolic protein 1 (LCP1 or l-plastin). lcp1 is expressed exclusively in hemopoietic cells, including macrophages. Macrophages are the targets of Mycobacterium tuberculosis, which is the etiologic agent of tuberculosis infection. LCP1 is an actin-binding protein theorized to influence macrophage motility. However, the role of LCP1 in tuberculosis pathology is not known. We present an infection method utilizing the zebrafish-Mycobacterium marinum model to study the role of lcp1 in TB. Recognizing the need for an infection model that is more reflective of the natural course of infection, we optimized a novel bath infection method for zebrafish larvae. This model mirrors human TB infections, with M. marinum localizing primarily in the gut. It successfully enables the colocalization of macrophages and bacteria seen via confocal microscopy, validating its potential for immune response analysis. We investigated the time and dose-dependent effects of bath infection on bacterial burden. Although inconclusive with limited samples, a trend of increasing bacterial burden over time was observed, while a dose-dependent effect was absent. The results allow for optimized experimental conditions to ensure valid observations. In conclusion, this infection model provides methods for better understanding TB pathogenesis, with the cultivation of the bath infection model offering promise for future infection studies. By offering valuable insights into the immune response and bacterial behavior, this model contributes to the hunt for improved TB treatments and prevention strategies. Future directions involve exploring ways to induce systematic infections in larvae and analyzing the genotypic response, allowing us to extend the model’s utility in advancing TB research.

Investigating the effects of juvenile hormone (JH) on behavior and morphology in adult bean beetles (Callosobruchus maculatus)

Raneem Samman ’24, Isabella Wojewski’24 and Flavia Barbosa, Biology Department, Lake Forest College, Lake Forest, IL 60045

Juvenile hormone is a sesquiterpenoid hormone produced and released by the corpus allatum (CA) in insects during their juvenile and adult stages. It is a critical regulator of both insect development and physiology. JH has previously been linked to resource allocation trade-offs between wings and gonads during the larval stage in bean beetles. However, the effects of JH on adult mating behavior are not fully understood. Building on this, we hypothesize that the effects of JH will depend on the timing of hormonal manipulation. Specifically, we predict that manipulations of JH during adulthood will modulate behavioral traits in adult males and females while exerting limited effects on morphological traits such as wing and gonad size. Upon emergence from pupae, we administered a JH analog (Methoprene) and a JH-synthesis inhibitor (Precocene) to both male and female bean beetles. We measured mating behaviors such as latency to copulate, male antennation rate, female latency to kick, and copulation duration. Concurrently, we measured gonad and wing sizes. This research aims to uncover the pivotal role of JH in shaping the behavior and morphology of adult bean beetles, while also shedding light on the hormonal mechanisms governing these processes and their broader implications in insect morphology and behavior.

Synthesis of Chaetoxanthone D with a Scandium Coupling Catalyst

Alexander Shanov ‘25, Leenah Ehsan ‘25, and Paul Gladen, Department of Chemistry, Lake Forest College, Lake Forest, IL, 60045

Xanthones are a structurally diverse natural product that come from the Chaetomium fungus. Chaetoxanthones have presented antiprotozoal, anticancer, and antimalarial properties from their natural counterparts. The bioactivity of the compounds is greatly influenced by their specific structure. Thus, we are developing a synthetic route to form Chaetoxanthone D to allow for further exploration into the biological activity. Past research was utilized in synthesizing the revised Chaetoxanthone D isomer by manipulating reaction conditions to minimize the formation of possible undesired products.

Reasonability of Force Assessments as a Mediator of the Link between Police Experience and Use-of-Force Decision Making

Zuzanna Smilnakova ‘24, Vivian P. Ta-Johnson, Brian Lande, Joel Suss, Amelie Motzer, Isabel Krupica, Sophie Rasof, Lauren Wright, Ceanna Loberg, Xianru Yu, Matias Fonolla, Wiktoria Pedryc, and Nilufar Immondodova, Psychology Department, Lake Forest College, Lake Forest IL 60045

Use-of-force decisions among police officers typically occur under stressful and fast-paced conditions. Because judgment and decision-making from more experienced individuals are considered more effective, efficient, and accurate compared to less experienced individuals, researchers have sought to understand the specific skills involved in the selection of appropriate uses-of-force among expert officers compared to novice officers. In the current study, we examined how a sample of expert (n = 42) and novice (n = 36) officers differ in their decisions in use-of-force scenarios. We also tested if officers’ justification of selecting a particular force option mediates the link between officer experience and force selection. Officers observed a series of body-worn camera footage of real-world police-citizen encounters across the US that were temporally occluded at several decision points. At these decision points, officers were prompted to describe the course of action they would take in the next few seconds if they were the officer on scene. Responses were coded for behaviors (n = 19) that officers indicated they would engage in. A series of linear mixed-effects models revealed that expert and novice officers differed on their decisions to (1) pursue a suspect; (2) deescalate the situation, and (3) subject themselves into a particular scenario. The results also indicate that officers’ reasonability of force assessments play a role in UoF selection differences between more and less experienced officers. Specifically, the increased odds of using less and lower levels of force among more experienced officers was largely mediated by officers’ assessment of the availability of force mitigation opportunities, nearby weapons/weapons of opportunity, and the suspect’s likelihood of escape were relevant in determining the appropriate level of force. Practical and research implications for expert performance in use-of-force training and decision-making are discussed.

Impact of a Four-Week Nutritional Summer Camp on Middle Schoolers’ Diets

Lauren Steidl ‘23, Niki Alyari, Ken Barber, and Dr. Kristin Schneider, Department of Psychology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Obesity resulting from poor diet is increasingly prevalent among children in America with nearly 20% of children between the ages of 2 and 19 having an obese BMI according to the CDC. Prevalence is especially high for children in low-income families that may find processed fast foods more affordable than fresh produce. Yet school gardening and nutrition programs have found success in improving children’s diets and reducing obesity by educating them on nutrition in a way that increases excitement about growing and cooking healthy foods. One such programs was introduced to John R. Lewis Middle School in Waukegan, Illinois, whose student body is composed mostly of racial and ethnic minorities, 95% of which qualify for free or reduced-price lunch programs based on family income. “Seed to Table” was a four-week summer camp for middle schoolers that aimed to teach students about the process through which we grow, harvest, and then prepare food. In addition to completing activities at the school’s greenhouse, students went on field trips to local farms and kitchens. Pre- and post-camp surveys were given to participating students in order to evaluate the effectiveness of this program at improving middle schoolers’ diets. Questions from validated adolescent nutrition measures, such as Project EATS Gen2 Survey and the CATCH Middle School Project, were used to measure change in processed food consumption, unprocessed food consumption, interest in cooking and nutrition, and engagement in camp activities. While the sample size was relatively small at only 13 students and the timeframe was a brief four weeks, the general trend of our results shows that continuation of such programs might have a positive impact on the nutritional health of middle schoolers.

Understanding the Structure of the preBötzinger Complex

Sara Taha ‘25, Babita Thadari and Kaiwen Kam, Department of Cell Biology and Anatomy, Rosalind Franklin University of Medicine and Science, North Chicago IL 60064

Breathing is an astonishing and crucial motor behavior that allows gas exchange through the lungs. The kernel for respiratory rhythm is found in a region of the medulla known as the preBötzinger Complex (preBötC), the borders of which are not well-defined.
The aim of this study was to determine the anatomical landmarks that identify the preBötC. It was hypothesized that examining the spatial distribution of medullary structures (i.e., facial nucleus, nucleus ambiguus, and inferior olives) would reveal the landmarks. To visualize neurons, thick medullary slices (550-650 μm) were generated from neonatal mice where preBötC had been localized electro-physiologically. Then, the boundaries of these structures were examined on the rostral and caudal sides of the slice in reference to preBötC. Most of the slices containing nucleus ambiguous were at a distance of ~200-300 μm from the facial nucleus. Hence, the preBötzingerC was near the nucleus ambiguus (NA), on the rostral side of the brain.

Subunit 4 of the ER membrane complex (EMC4) plays a key role in cells of the nervous system

Otoha Tatami ’24, Inés Riojas ’25, Maria Jose Orozco Fuentes ’24 and Dr. Rebecca Delventhal Ph.D, Department of Biology and Neuroscience, Lake Forest College, Lake Forest, IL 60045

The endoplasmic reticulum (ER) is involved in the modification, packaging, and insertion of membrane proteins in the cell. The ER membrane protein complex (EMC) is composed of 8-10 subunits that work together to facilitate protein biogenesis. Through a previous random RNAi screen, our lab observed that fruit flies (Drosophila melanogaster) without the EMC subunit 4 in glial cells resulted in severe phenotypes, such as delayed development and a very short lifespan of 5-6 days instead of the normal 2-3 months. From this, we investigated the role of EMC4 in other cells of the fly’s brain and body. We knocked down expression of EMC4 in all neurons, and subtypes of neurons, such as insulin-producing and circadian regulatory neurons. Knockdown of EMC4 in all neurons and just insulin-producing neurons led to developmental delay, similar to what was observed in glia. Knockdown of EMC4 in circadian regulatory neurons led to a shorter lifespan of 23-28 days, similar to knockdown in glia, but not as severe. These findings suggest that EMC4 plays a key role in the cells of the nervous system. Additionally, we investigated the effect of the EMC4 knockdown during the developmental stages or adulthood by restricting the knockdown timing. We found that restricting the knockdown of EMC4 to adulthood resulted in a mild reduction in lifespan relative to controls, but not as severe as observed with the continual knockdown, suggesting a more important role of EMC4 in glia during development. Future research will focus on the behavioral effects of EMC4 knockdowns during the larval stage as well as the impact of temporally restricted EMC4 knockdowns in all neurons and insulin-producing neurons. Overall, our research highlights the importance of better understanding the role of EMC in producing membrane proteins, which are crucial for many aspects of the nervous system.

Telomerase Levels Dramatically Increase in Ascospores

Petra Urgacova ‘25, Meklit Yimenu ‘23, and Karen Kirk, Biology Department, Lake Forest College, Lake Forest, IL 60045 

Telomeres are repetitive and noncoding sequences at chromosome ends that prevent DNA loss during replication. The primary enzyme responsible for telomere replication is telomerase. Telomerase is a ribonucleoprotein with two essential components, TER and TERT. TER is an RNA that contains the template for repetitive telomere synthesis, and TERT is a reverse transcriptase. Assembly of the two components occurs in the cytoplasm in Saccharomyces cerevisiae and in the nucleus in mammalian cells. The structure of TER in the filamentous fungus Aspergillus nidulans is partially conserved in both S. cerevisiae and mammalian cells but whether TER migrates to the cytoplasm or remains in the nucleus during telomerase assembly was unknown. We used the heterokaryon rescue technique in A. nidulans hyphae and found that TER remains in the nucleus and, thus, telomerase assembly occurs here. To further investigate this observation, we wanted to explore the expression levels of the TER and TERT genes at various life cycle stages. Nearly ten replications of RT-qPCR were used to determine expression levels in muti-nucleate hyphae, asexual conidiospores, sexual ascospores, germinating conidiospores, and germinating ascospores. Surprisingly, we found that the relative expression levels of TER and TERT were at least two-fold higher in the germinating ascospores compared to hyphae, even though, based on PCR, the telomere lengths were similar. We also looked at expression levels in nkuA and POT1 mutant cells. We used these cells because nkuA and POT1 are involved in nonhomologous end joining and telomere protection, respectively, and because we wanted to test whether these mutations affected expression levels in the different cell types. The relative expression levels of TER and TERT were also higher in the germinating ascospores in these mutant cells. Hence, TER and TERT have higher expression in ascospores in both wild-type and mutant A. nidulans cells. These results imply not only the upregulation of TER and TERT but possibly longer telomeres in the sexual spores.

Sex differences in cognitive decline, affective behaviors, and PV interneuron activation in AD mice after isolation

Megha Verma ‘25, Lainey Toennies, Kameron Kaplan, Nicole Ferrara, and Holly Hunsberger, Center for Neurodegenerative Diseases and Therapeutics, Stress Center: Center for Neurobiology of Stress Resilience and Psychiatric Disorders, Rosalind Franklin University of Medicine and Science, North Chicago, IL.

As innately social beings, humans are motivated by and dependent on social relationships throughout their lifetime. The importance of social interaction is emphasized by studies that demonstrate social isolation as a risk factor for negative health outcomes such as cardiovascular disease, increased risk for mortality, depression and anxiety, and cognitive decline. Elderly people are particularly susceptible to becoming isolated and cognitive decline is higher among seniors who report loneliness, and this is greater among females. In contrast, intact social networks are highly protective against dementia in both humans and rodents. While there are familial AD mutations, most cases of AD are sporadic, and the cause is unknown. However, one promising avenue to reduce AD prevalence by 4% is to eliminate social isolation as a risk factor. This would provide a greater reduction in prevalence than combatting physical activity (2%) and hypertension (2%). Moreover, the recent COVID-19 pandemic forced millions into repeated social isolation periods ranging from several weeks to months, a situation which could occur in the future. Therefore, it is essential to understand risk factors, such as isolation, for early interventions. The goal of this proposal is to understand the sex-specific effects of social isolation on memory and social interaction in aging and AD mice. Our central hypothesis is that social isolation alters both long-term memory and social behavior to a greater extent in AD phenotypes that will be differentially affected between sexes. To explore the impact of social isolation on AD, we ran a battery of social, anxiety, and cognitive behavioral tests. Although most isolation studies result in memory impairment, our AD female mice exhibited enhanced memory retention after acute isolation as measured by contextual fear conditioning. Isolated male control and AD mice showed a memory deficit compared to group housed mice. Social interaction was decreased after a stressor in all groups, and this was exacerbated by isolation in female mice. We are currently examining brain tissue to determine excitatory/inhibitory cell activation using cFos, an immediate early gene, and Parvalbumin (PV), an inhibitory interneuron. We predict increased cFos and decreased PV activation in isolated AD females, a disturbance in the excitation/inhibition balance in the amygdala and hippocampal regions. The results from the proposed project would provide foundational evidence for the impact of the social environment on age-related cognitive decline. These results would mechanistically identify a novel pathway through which social circumstances impact degenerating neural circuits to drive age-related alterations in behavior.

Advancements in Gene Therapy for Alzheimer's Disease: Brain-Targeted Inhibitors in Mouse Models

Lira Zajmi ‘25, Afrah Rehman, Mike Milovanovic, and Dr.Robert Marr, Department of Neuroscience, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60045

Age-associated dementias, like Alzheimer's disease (AD), have significant consequences as the elderly population grows. Conventional drug development for AD has faced challenges, necessitating alternative approaches. In this research, we investigated the role of inflammation in AD, focusing on the NLRP3 inflammasome complex, a promoter of AD pathology. Inhibiting NLRP3 has shown promise in protecting against AD in mouse models. Our research explores gene therapy using adeno-associated virus (AAV) vectors to express a secreted dominant negative inflammasome inhibitor, offering the potential for widespread delivery. We present results from our study, demonstrating the successful expression of the secreted product from the AAV vector in cell culture, confirming the vector's functionality. Additionally, we engineered a lentiviral vector expressing the blood-brain barrier (BBB) crossing recombinant inhibitor. Subsequently, we also confirmed the secretion of this new product in cell culture. This research will increase our understanding of the role of the inflammasome and anti-inflammatory treatments for Alzheimer's disease.