Latent class analysis of a heterogeneous international sample of patients with ME/CFS
Kayla A. Huber, Madison Sunnquist, & Leonard A. Jason. DePaul University, Center for Community Research
Individuals with myalgic encephalomyelitis (ME), or chronic fatigue syndrome (CFS), routinely display differences in symptomatology, as well as illness course, onset, duration, and functional disability. In an attempt to elucidate potential subtypes of the illness, twelve less frequently-endorsed symptoms were included in a latent class analysis of 1,210 adults with ME or CFS. The resulting six-class solution consisted of one class that was likely to endorse all atypical symptoms, one class that was unlikely to endorse any atypical symptoms, and four classes that were likely to endorse either one or two atypical symptom domains. An exploratory multinomial logistic regression indicated that education, work status, illness course, and how long ago fatigue began were significant predictors of class membership. These results are suggestive of subtypes of ME and CFS and, if replicated, may assist physicians in providing tailored treatment to patients and allow researchers to form more homogeneous samples.
Understanding Compensatory Mechanisms in a Dopamine Deficient Mouse Model: Implications in Parkinson’s Disease
Sierra Smith, Poulin JF., Wang YZ., Savas J., Awatramani R. Department of Neurology, Northwestern University, Chicago IL, 60611
Parkinson’s Disease (PD) is a progressive neurodegenerative disease characterized by neuronal degradation, dopamine (DA) deficiency in the midbrain striatum, and motor dysfunction. Interestingly, motor symptoms of PD only begin to show after 60-80% of DA neurons have been lost. In PD patients, DA levels are restored by treatment with L-DOPA, the rate-limiting enzyme in DA production. For the long pre-symptomatic period, compensatory mechanisms can delay the onset of motor symptoms. It is not currently well understood how these mechanisms function at the molecular level. The goal of this project is to understand these compensatory changes at the molecular level in dopamine deficient mice as they relate to PD. A proteomics study of the striatum in a novel conditional Th knockout mice revealed a set of proteins that are either upregulated, downregulated, or have no significant change in expression. One pathway in particular showed multiple proteins upregulated significantly, the mitogen activated protein kinase (MAPK) pathway which is involved in many cellular processes including regulation of transcription and translation. We hypothesize that the MAPK pathway is overactive in a compensatory manner during the pre-symptomatic stage in PD and that agonizing this pathway during disease progression may have therapeutic benefits.
Perceptions and Barriers to Breastfeeding in African American (AA) Women at an Urban Chicago Hospital.
Jessica A. Dudley MS, RN. University of Illinois at Chicago | College of Nursing, Chicago, IL 60612
Intro/Background: Breastmilk is known as “superior nutrition” that confers benefits to both mother and child. However, African American (AA) women are least likely to breastfeed, with about 66.3 percent of AA mothers initiating breastfeeding compared to an 81.1 percent national average. Aim: This project aims to understand the perceptions that AA mothers on the postpartum unit at the University of Illinois Hospital (UIH) have towards breastfeeding. Methods: A modified paper version of the Iowa Infant Feeding Attitude Scale was created, and surveys were distributed to mothers on the postpartum unit of the hospital during lactation rounds. Mothers on the labor and delivery unit and pregnant mothers on the antepartum unit were excluded from the study. Twenty-nine items to measure maternal attitudes towards various feeding methods were divided and compared between AA women and Non-AA women. Results/Conclusions: AA women (compared to non-AA women) at UIH tend to have generally positive perceptions of breastfeeding, with fifty percent of women planning to breastfeed exclusively, and an even higher percentage of AA women planning to exclusively breastfeed for more than six months. Implications/Limitations: AA women generally have a plan to breastfeed; however, national trends show problems with initiation and continuation. Therefore, more support from Lactation nurses may be needed. Future studies may look into the history of breastfeeding and media contributions (e.g. baby dolls being sold with bottles) to breastfeeding perceptions. Limitations of this study included a limited sample size and limited analysis of items on questionnaires distributed to patients.
Characterization of cell proliferation and palate shelf elevation in Prdm16 null mutant embryos.
Sahishnu Patel1, Cosmo Vivirito2, Charlotte Imlach3, Angela Gomez1, Bryan C. Bjork1. Midwestern University, 1Chicago College of Osteopathic Medicine, Dept. of Biochemistry and College of 2Health Sciences, Masters of Arts in Biomedical Sciences Program; 3Cornell College, Mount Vernon, IA.
Cleft lip and palate is one of the most common human birth defects, and understanding its underlying developmental, genetic and cellular disturbances will eventually improve its prevention, treatment, and prognosis. Prdm16 hypomorphic mouse mutants exhibit cleft palate (CP) due to failed palate shelf elevation as a secondary consequence of micrognathia, which models human Pierre Robin sequence cleft palate. This study aims to evaluate cell proliferation in the developing mandible and palate in Prdm16 gene trap null mutant mice to determine if complete loss of Prdm16 function will uncover a palate-intrinsic role for Prdm16 in addition to it palate-extrinsic role. We hypothesized that Prdm16 null mutants will exhibit altered cell proliferation rates in the developing palate and that they will demonstrate a lower rate of palate fusion in suspension palate cultures with mandibles removed. A comprehensive assessment of cell count and cell proliferation rates in wt and Prdm16 mutant palate shelves through immunofluorescence using Anti-Phospho-Histone H3 labeling did not uncover differences in cell proliferation overall, although we did observe significant differences in a small number of the regions evaluated. We observed that mutant palate shelves had increased proliferation in the medial aspect of the anterior palate, both medially and laterally in the middle palate, and in the lateral posterior palate. Further, we identified an increase in the total number of cells in the perichondria of Prdm16 mutants and a corresponding decrease in Meckel’s cartilage (MC) cell count. No significant difference in MC cell proliferation was observed. In vitro suspension palate cultures further elucidated that Prdm16 mutant palate shelves are able to fuse in the absence of a mandible, thus disproving our hypothesis that the null mutation would lead to phenotypic expression intrinsic to palate shelves. These data remain consistent with our past observations that cleft palate occurring with Prdm16 loss is secondary to palate-extrinsic factors, specifically micrognathia, which physically obstructs palate shelf elevation.
Corticotropin-releasing factor (CRF) neurons in the oval nucleus of the bed nucleus of the stria terminalis (BNSTov) modulate fear and anxiety in rats
Alexandra N. Roman, D. Martinon, and J. Dabrowska. Cellular and Molecular Pharmacology and Neuroscience Departments, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
Corticotropin-releasing factor (CRF) is a neuropeptide that regulates the autonomic, endocrine, and behavioral responses to stress. One significant population of CRF cell bodies is located within the oval nucleus of the bed nucleus of the stria terminalis (BNSTov), a region of the brain that mediates adaptive responses to stressors. This research investigates the role of BNSTov CRF neurons regarding the acquisition of conditioned cued and non-cued fear. We modulated this system using designer receptors exclusively activated by designer drugs (DREADDs) within adult transgenic CRF-Cre rats (developed by Pomrenze et al., 2015). In this model, Cre recombinase is exclusively expressed by CRF neurons. We performed bilateral BNSTov injections of viral vectors driving Cre-dependent expression of DREADDs and reporter protein, mCherry, in both Cre+ and Cre- rats. Viruses used encoded either DREADDs-Gi (pAAV-hSyn-DIO-hM4D(Gi)-mCherry(AAV8)) to silence CRF neurons or DREADDs-Gq (pAAV-hSyn-DIO-hM43(Gq)-mCherry(AAV8)) to activate them. Four weeks after injections, we measured baseline acoustic startle reactivity of all rats. Then, we used a fear-potentiated startle (FPS) paradigm to determine if activating or inhibiting BNSTov CRF neurons affects cued and non-cued fear. The DREADDs selective ligand, clozapine-N-oxide (CNO, 1 mg/kg/mL, IP), was administered prior to fear conditioning. Rats were tested for FPS expression 24 hours later. To confirm DREADDs expression, we used striatal-enriched protein tyrosine phosphatase (STEP), which selectively colocalizes with BNSTov CRF neurons. We confirmed through fluorescent immunohistochemistry that mCherry was exclusively expressed on CRF neurons in the BNSTov. We also found that activating or inhibiting CRF neurons in the BNSTov modulates cued and non-cued fear measured in FPS in distinct ways. In light of this data, future experiments will aim to further elucidate the functional role of CRF neurons in the BNSTov and confirm the specificity of DREADDs expression in this promising animal model. This work is supported by a National Institute of Mental Health grant (R00MH096746) and CMS RFUMS start-up funds to JD.
Paired recordings of pyramidal cells in the subiculum reveal local excitatory microcircuits
Michael Fiske and G. Maccaferri. Department of Physiology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611; Northwestern University Interdepartmental Neuroscience Program, Northwestern University, Chicago, IL 60611
Information exchange between neurons is accomplished using sequences of action potentials that result from the integration of local microcircuits. Unraveling the connectivity of these microcircuits and how they contribute to network activity is vital for understanding how information is relayed through the brain. Interestingly, despite its role as the main output region of the hippocampus, the microcircuity of the subiculum remains understudied. Additionally, recent evidence suggests that the subiculum is involved in generating both interictal and ictal activity in epileptic patients, providing impetus to study how these microcircuits contribute to disease. Most work involving the subiculum has focused on the excitable properties of the constituent pyramidal cells, which can be classified as either regular spiking or bursting. However, little is known about the local synaptic connectivity between these distinct neuron types. My project aims to physiologically and anatomically characterize the excitatory connections of the subiculum at the individual neuron level. Using simultaneous, triple whole-cell recordings, I have shown significant levels of connectivity between the principal cells of the subiculum (n=1828 tested connections). Connections were observed between bursting to bursting, regular to regular, bursting to regular, and regular to bursting neurons. These synaptic connections are excitatory and mediated by AMPA receptors at resting potential. Future experiments will evaluate the kinetics of the recorded responses, and assess the contribution of NMDA receptors to these synaptic connections. Additionally, anatomical reconstruction of recorded cells will allow me to map the location of putative synapses in recorded pairs. Accumulating evidence has also implicated excitatory GABAergic signaling, resulting from decreased expression of the KCC2 chloride transporter, in generating population activity linked to seizures. My preliminary experiments show that a KCC2 transporter antagonist triggers synchronous depolarizations in the subiculum, even when outside inputs are disrupted. Ultimately, this work will provide insight into the population dynamics of the subiculum, which is vital for understanding the normal physiology of the subiculum, as well as its role in epilepsy.
Identifying sex differences in the rat basolateral amygdala to the central amygdala circuitry through in vivo electrophysiological stimulation
Brittany Avonts, J. Vantrease, A. Rosenkranz. Department of Cellular and Molecular Pharmacology, Rosalind Franklin University, North Chicago, 60064; Department of Neuroscience, Lake Forest College, Lake Forest, IL, 60045
Anxiety disorders are the most common type of psychiatric disorders with the prevalence two times higher among females than males. The amygdala is hyperactive in patients with anxiety disorders and has sex differences in activation in response to emotional provoking stimuli. The basolateral amygdala (BLA) is known to play a role in fear and anxiety and our lab has previously shown that BLA activity in naïve female rats is increased compared to naïve male rats. The BLA has many downstream projections, so we specifically wanted to look at the projection to the central amygdala (CeA), which is necessary for cued fear conditioning and has connections controlling the motor expression associated with fear behaviors, such as freezing. Sex differences have also been observed in behavioral responses to cued fear, with females freezing more than males in response to a conditioned tone. Since the CeA is required for cue conditioned freezing, we hypothesized that there will be a stronger output. in females. In order to test this hypothesis, we used single unit in vivo electrophysiology and stimulated the CeA to antidromically identify BLA neurons that project to the CeA. Understanding the specific outputs of the BLA could lead to novel target therapies.
Starved of Oxygen: The insulin regulation of growth in response to hypoxia in Drosophila
Sam Gascoigne and A. W. Shingleton. Department of Biology, Lake Forest College, IL 60045
In almost all animals, a lack of oxygen during development increases developmental time and reduces growth rate and final body size. Despite the ubiquity of this phenomenon, the mechanisms that underpin a hypoxic response are largely unknown. Using the fruit fly Drosophila melanogaster as a model organism, we have shown previously that the steroid hormone ecdysone, which inhibits growth, is necessary for a hypoxic response. In addition, we have shown that hypoxia inhibits insulin/insulin-like growth factor signaling (IIS). Since ecdysone also inhibits IIS, we therefore tested whether a functional IIS is necessary for flies to mount a hypoxic response. Our data show that IIS-mutant flies do not reduce their growth rate or increase their developmental time in hypoxia. Collectively, our data suggest that the hypoxic response is mediated through the IIS in Drosophila.
Evaluation of the Hippocampus After Repeated Concussive Traumatic Brain Injury (TBI) at a Chronic Time Point in Rats
Danielle J. Sychowski, E. Reisenbigler, J. Love, M. Thaqi, and D. A. Peterson. Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
Mild forms of traumatic brain injury (TBI), such as concussions from falls, sports, or combat, are often associated with memory deficits that may or may not be reversible. Even a single TBI can alter the brain in such a way that leaves it vulnerable to further injury. Experiencing multiple TBIs has been shown to increase risk of neurodegenerative disease, particularly chronic traumatic encephalopathy (CTE), which has been increasingly found in the brains of former American football players. We recently described a clinically relevant closed-head model for mild TBI, and are using this model in rats to evaluate mechanisms of neural injury and to explore possible therapeutic strategies. Our analysis is focused upon injury in the hippocampus of rats that underwent mild repeated TBI at a chronic time point. Sagittal sections of the brains were immunostained to detect mature neurons (NeuN), early neuroblasts (DCX), and proliferating cells (BrdU). These sections are being assessed by confocal microscopy to assess neuronal loss and impact upon proliferating cells in the hippocampus. Darkfield images throughout the entire hippocampus are being measured by stereology (Cavalieri Estimator) to establish the extent of changes in brain volume. Establishing neuronal loss in the hippocampus would provide a useful model for investigating regenerative approaches to replacing neurons through stimulating adult neurogenesis in the hippocampal dentate gyrus or reprogramming induced neurons in the hippocampal CA subfields.
Mild Chronic Controlled Cortical Impact in Alzheimer’s-like Mice
Trevor Buhr, Kathleen Maigler, and Robert Marr Department of Neuroscience, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, IL
Traumatic brain injury (TBI) has become a fast-growing area of study as more athletes and young people have been documented with cognitive deficits after sustaining repeated head injuries. Previous research has shown that repeated traumatic brain injury can result in an increased risk of dementia and diseases such as chronic traumatic encephalopathy (CTE). Although how these diseases manifest after head injury is not well studied. Models are well-developed in rats and in mice using open-skull impacts, however, we attempt to establish a more clinically relevant model of repeated head injury in mice using repeated mild closed-skull controlled cortical impact. Our model examines mice with Alzheimer’s Disease-like pathology by transgenic expression of the mutant human Amyloid Precursor Protein (APP) gene as well as mice bred with the human wild-type form of APP. Using this clinically relevant method of disease and injury, we expect to uncover the acute effects of Amyloid-beta in TBI pathology. Behavioral tasks such as Morris water maze and novel object recognition tests as well as rotarod assessment will be used to determine whether mice have received TBI. Brain dissection and tissue staining will then be used to examine the brain pathology of TBI and discern the role of Amyloid-beta and enzymes involved in its clearance in chronic mild TBI.
Effects of autonomic arousal on math performance in children with and without ADHD
Krista Meuli, R. Gyorfi, and N. Wentworth. Department of Psychology and Neuroscience Program, Lake Forest College, IL 60045
Attention Deficit/ Hyperactivity Disorder (ADHD) is one of the most prevalent neurodevelopmental disorders, affecting 1 in 20 U.S. children. ADHD is characterized by chronic inattention, hyperactivity, and impulsivity. Though ADHD is a well-studied disorder, the underlying attention deficits in ADHD remain poorly understood. Previous research on attention and task engagement has found pupil dilation, an autonomic nervous system response of arousal, to be an effective measure of fluctuations in mental effort related to arousal, such as attention. One area of ADHD research has investigated decreases in academic performance observed in children with ADHD. Underperformance in mathematics, particularly, has been described in children with ADHD compared to typically developing peers. However, the role attention deficits play in this decreased math performance is still unknown. This study investigates how fluctuations in autonomic arousal correlate with mental arithmetic performance in children with and without ADHD. First, a mental arithmetic task with 40 basic addition problems was developed. Participants ages 9-12 in either the ADHD group or typically developing group will be given the time-constrained mental addition task, while recording pupil dilation at 60 Hz and latency to answer. Between each problem, a baseline of pupil dilation will be recorded. Differences in pupil dilation patterns and latency to answer on correctly and incorrectly answered problems will be compared, as well as patterns between ADHD and typically developing groups.
Differential impact of several PD-associated genes on the toxicity associated with wild-type and familial mutants forms of α-synuclein in a yeast model
Paul Jones, Emily Ong, Ariane Balaram, Alexsandra Biel, Chisomo Mwale, Maiwase Tembo, and Shubhik DebBurman. Neuroscience Program and Biology Department, Lake Forest College, IL 60045
Parkinson’s disease (PD) is a neurodegenerative disease characterized by motor impairment. The aggregation of the α-synuclein in the midbrain dopaminergic neurons and the death of these cells underlie these symptoms. PD can be both genetic and sporadic. Familial PD is directly caused by a mutation in one of at least 10 genes, including α-synuclein, DJ1, Vps35, and ATP13A2. α-Synuclein has six identified missense mutations (A30P, E46K, H50Q, G51D, A53E, and A53T) that each cause early-onset PD. Sporadic PD is linked with several risk genes, including Vps13, Sac I, and Swa2. In our budding yeast model system, we find wild-type, E46K, A53T, H50Q, and A53E α-synuclein to be toxic to yeast and they show varying degrees of membrane binding and aggregation, while A30P and G51D α-synuclein are relatively non-toxic and show cytoplasmic diffuse localization. What is still not well understood is whether the PD-causing and PD-risk genes mentioned above can influence the toxicity properties six familial mutants of α-synuclein. To explore this connection, wild-type and familial forms α-synuclein were studied in yeast strains that were singly deleted for these six PD-linked genes. Results indicate that some gene deletions increased α-synuclein toxicity (DJ1, VPS13), others reduced toxicity (ATP13A2, VPS35), and some have no effect (Swa2, Sac1). In each case, those genes that altered α-synuclein toxicity did so in a familial mutation-specific way. Our findings suggest that each familial mutant creates cellular toxicity in a unique way regulated by different subsets of genes, opening doors for mechanistic insight into the mode of toxicity with each mutant.
α-Synuclein expression in YCLN-3 yeast strains demonstrate differentiated response patterns under various conditions of stress
Tom Steen and David Mueller. Department of Biochemistry and Molecular Biology, Rosalind Franklin University Medical School
Many years of cellular and molecular research has allowed us to perform experiments with efficiency and ease. Yeast has proven to be one of the most effective model organisms to utilize when analyzing protein level interactions. In these experiments, isolations and transformations were done on the yeast strain DMY929 with a null mutation in the yeast homolog (YHC3) for the human gene (Cln3) defective in Batten disease. Plasmids expressing the Alzheimer’s associated protein α-Synuclein were introduced into yeast DMY929, YPH 499, and LWY 078. We examined the effect of expression of α-Synuclein and α-Synuclein with the pathogenic, A53T mutation in the daughter strains under stressors of heat and different “food” conditions. Growth of each strain and mutation was analyzed qualitatively through dot assays and quantitatively through growth curves. These experiments uncovered that the stressor of heat hindered the growth of the A53T mutations independent of the daughter strain. Our results also demonstrated that SGAL (a galactose medium) hinders the growth of the α-synuclein mutations.
Exploring the Role of Specific Amino Acids in Protein Aggregation: Site-Directed Mutagenesis of the Swi1 Prion Domain
Sarah V. Applebey, D. K Goncharoff, and L. Li. Department of Biochemistry and Molecular Genetics, Feinberg School of Medicine, Northwestern University, Chicago IL 60611
Prion are proteins that have the ability to misfold into an alternative, infectious conformation. These alternative conformations cause diseases such as Creutzfeldt-Jacob and transmissible spongiform encephalopathies. In addition, proteins that play a role in neurodegenerative diseases have been shown to propagate in a prion-like fashion, such as, β-amyloid and tau in Alzheimer’s disease and α-synuclein in Parkinson’s disease. Using the budding yeast Saccharomyces cerevisiae as a model organism, this project sought to elucidate the effects of specific amino acids on prion formation, maintenance, and propagation. This was done by manipulating the prion domain of endogenous protein Swi1, which can misfold into the prion conformation [SWI+]. Through site-directed mutagenesis, we analyzed the ability of several Swi1 variants to aggregate with different levels of Swi1FL (full-length) expression and maintain [SWI+] in the absence of Swi1FL. Preliminary results implicate the importance of phenylalanine residues in aggregation, as mutants without phenylamine residues do not maintain the prion. The replacement of threonine repeats with alanine, but not asparagine produced a reduction in prion expression. However, singular threonine residues were not required for prion formation. To confirm these findings, future research will include western blots of mutants and analysis additional Swi1 variants
Exon Skipping for the Therapeutic Treatment of Juvenile Batten Disease
Grant Brady, J. Centa, and M. L. Hastings. Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
Batten disease is a fatal neurodegenerative disease in the neuronal ceroid lipofuscinoses (NCL) family. Disorders in this family are characterized by excessive accumulations of lipopigments in body tissues, including the brain. Juvenile Batten disease is the most common type of NCL. Symptoms usually arise between the ages of 4 and 8, and include vision loss, seizures, loss of motor function, personality change, and overall cognitive decline. Juvenile Batten disease is most commonly caused by the deletion of exons 7 and 8 in the CLN3 gene. The mutation shifts the reading frame of the mRNA leading to a premature stop codon resulting in a nonfunctional protein. There is currently no effective therapy or treatment for Batten disease. We hypothesize that correcting the reading frame by deleting exon 5, 6, or 9 will restore CLN3 protein function. By comparing the wildtype, CLN3∆ex7/8, and the corrected reading frame isoforms CLN3 ∆ex5/7/8, CLN3 ∆ex6/7/8, and CLN3 ∆ex7/8/9, we are working to determine whether or not deleting an extra exon on top of the mutation deletion of exons 7 and 8 will be therapeutically beneficial. In this study, we used cell lines, yeast, and CLN3 mouse models to compare the characteristics of the different isoforms of the CLN3 gene. In addition, we are trying to determine the overall function of CLN3, since it is still not clear. We are looking at where CLN3 is localized in cells, as well as growth phenotypes in yeast via drop testing to see if CLN3 is needed for growth at a higher temperature. Seizure susceptibility testing in mice can also help determine a possible function. CLN3 ∆ex7/8 mice are more susceptible to pharmacologically induced seizures compared to the wildtype, suggesting that CLN3 may have a role in reducing seizure activity in the wildtype mice. These data improve the knowledge of the functionality of CLN3, as well as assess the therapeutic strategy of exon skipping in Juvenile Batten disease.
The role of nucleolin in inflammatory breast cancer and in Kaposi’s sarcoma-associated virus malignancies
Sandra P. Campos, M. Repak, N. Sharma-Walia. Department of Microbiology and Immunology, Rosalind Franklin University of Medicine and Science, Chicago, IL
Nucleolin is a type of protein found to be involved in cell proliferation and cell death in many different types of cancers including inflammatory breast cancer and primary effusion lymphoma. Inflammatory breast cancer is a rare type of breast cancer characterized by redness and swelling of the skin around the breast. Diagnosis of inflammatory breast cancer can be difficult and further research is needed to clearly understand this disease. Primary effusion lymphoma, also known as B-cell body cavity lymphoma (BCBL), is a rare type of Kaposi’s sarcoma-associated herpesvirus (KSHV) malignancy. Patients have poor prognosis, and they usually have about 6 months left to live after diagnosis. Previous studies of mass spectrometry data from our lab showed nucleolin to a binding partner of osteoprotegerin (OPG) in inflammatory breast cancer cell lines. Human mammary epithelial cells (HMEC) were used as non-IBC control cells. Inflammatory breast cancer cells (SUM149PT, SUM190PT) were used as IBC cell lines. Nucleolin expression was found to be higher in IBC cell lines SUM149PT, SUM190PT, and MDA IBC-3, in comparison to control HMEC cells. Previous studies of immunoprecipitation from our lab showed nucleolin to be expressed in KSHV-positive cells, with the virus in the lytic stage. BJAB cells were used as KSHV-negative control cells and BCBL cells were used as KSHV-positive cell lines. Nucleolin was found to be expressed more in the cytoplasm of KSHV-positive cells, than in the KSHV-negative cells. By examining the role of nucleolin on the apoptotic pathway, we will get a molecular understanding of its function in cell death in inflammatory breast cancer and in primary effusion lymphoma. Understanding the functionality of nucleolin in the signaling pathways of inflammation in breast cancer scenario will provide new avenues of promising therapeutic targets.
Perineuronal Net Expression in Amygdala
Michael Buabeng, M. Bompolaki, and J. Urban. Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
Stress resiliency is the ability of an individual to successfully adapt to an environment with adverse conditions. Previous studies show that the amygdala is associated with stress, and that neuropeptide Y (NPY) promotes stress resilience through actions in the basolateral amygdala (BLA) yet a mechanism has not been defined. Perineuronal nets are extracellular matrices surrounding neurons, predominantly parvalbumin interneurons, to stabilize cellular activity. Therefore, we hypothesized that there will be an increase in the expression of cells expressing perineuronal nets in in the basolateral amygdala induced by NPY. To test this theory, we sliced sections of brains from vehicle- and NPY-treated rats, our model organism, containing the amygdala: lateral and basolateral regions. Then, we did double-labeling immunohistochemistry to detect the parvalbumin cells and wisteria floribunda agglutinin (WFA) to detect the perineuronal nets sections of the brain were atlas-matched to standardize the area of the amygdala and the number of WFA-positive cells were counted unilaterally. There was a strong association of WFA with parvalbumin neurons as determined previously. However, there were fewer WFA-positive neurons in the lateral amygdala compared with the basolateral aspect. While there were overall more WFA-positive cells in the BLA of NPY-treated animals, this was not statistically significant. Currently, we are adding more animals to the groups to strengthen the analysis. These data will be important to determine possible mechanisms in the amygdala which support the development of resilience.
The Effects of 5-Azacytidine and Reversine treatment on Muscle Fiber Formation in Cerebral Palsy.
Johanna E. Hendriksen1,3, A. M. Taylor2,3, A. Domenighetti, Ph.D3.1 Lake Forest College, Lake Forest, IL; 2 Alabama College of Osteopathic Medicine; 3 Shirley Ryan Ability Lab, Chicago, IL
Cerebral palsy (CP) is the most common group of childhood disability. It is caused by a non-progressive perinatal brain lesion. The impact of the brain lesion leads to progressively shorter, thinner, and contracted skeletal muscles during postnatal development. We wanted to test two drugs, 5-Azacytidine and Reversine, to see if they had restorative properties on muscle growth in CP. We hypothesized that 5-Azacytidine, with or without Reversine, would trigger muscle stem cells (MuSCs) to restore the size and number of muscle fibers in CP to typically developing (TD) levels. MuSCs were isolated from muscle biopsies obtained from CP or TD children and differentiated into muscle fibers in culture. We cultured MuSCs under multiple conditions, including control, 6 or 24 hours treatment with 5-Azacytidine, 3-day treatment with Reversine, or 24 hours co-treatment with 5-Azacytidine and Reversine followed by 2 days with Reversine. The Reversine and 5-Azacytidine co-treatment proved the most favorable in restoring muscle fiber growth to TD levels in CP. Our results will assist in the development of drug therapies that may support muscle rehabilitation in children with CP.
Effect of removable cast walkers’ size and use of a contralateral lift on postural stability of diabetic neuropathic patients
Jacob Tilsley B.S.1, Abagayle King2, Ryan Crews MS, CCRP1. 1Center for Lower Extremity Ambulatory Research (CLEAR), Dr. William M. Scholl College of Podiatric Medicine at Rosalind Franklin University of Medicine and Science; 2Lake Forest College
Diabetes mellitus affects one of every eleven adults, and each patient has a 25 percent risk of developing a diabetic foot ulcer (DFU) over their lifetime. The primary treatment for DFUs includes reducing plantar pressures (offloading) using a removable cast walker (RCW); however, compliance to these devices are poor. These devices are generally large, heavy and create a limb length discrepancy. Previous research has shown self-reported postural instability is predictive of RCW adherence levels. Prior to this study, investigations concerning RCWs and postural stability have generally been limited to assessing body sway during quiet standing. This study seeks to determine if different offloading options impacted users’ compensatory responses to perturbations as well as whether the different offloading options influenced users’ ability to avoid obstacles while walking. In this study, we are testing three conditions (bilateral standard shoes, knee-high RCW and contralateral standard shoe, ankle-high RCW and contralateral standard shoe with external lift) to determine if postural stability is affected by the varied offloading options. Data has been collected on the first three patients to date. Presently there is a trend indicating the ankle-high RCW and contralateral lift make stability better than a traditional knee-high RCW. Participants also report feeling more stable in the ankle-high RCW and contralateral lift than in the knee-high RCW. Should this trend hold once full enrollment is met (25 subjects) the results will indicate the ankle-high RCW and contralateral lift are likely to yield greater adherence levels by individuals with active DFU than a traditional knee-high RCW.
Mutating the Telomere Gene
Anna Sandler and Brett Palmero. Department of Biology, Lake Forest College, IL.
Telomeres are non-coding nucleotide sequences at the ends of chromosomes and serve to prevent the loss of genetic information during cell replication. To prevent telomeres from shortening, a telomerase complex consisting of a protein (TERT) and an RNA template (TER) helps maintain the length of the telomeres. Crucial to the success of telomerase is its regulation. Downregulation of telomerase is the basis of aging while upregulation of telomerase can result in cancerous cells. Our lab has been studying the biogenesis, or synthesis and localization, of telomerase. Previous research in our lab studied the localization of TER in Aspergillus nidulans, a species of fungi that can exhibit a multi-nucleate state known as a heterokaryon and can help determine if a gene is essential. Researchers in our lab knocked out the gene for TER to determine if it leaves the nucleus, and initial results suggest that it does not. In our project, we sought to knockout the TERT gene by creating a three-piece knockout construct that would ultimately be used to test the localization of the TERT gene product. We designed primers to amplify the fragments and ultimately fuse them together to create the construct. Although we successfully amplified the three pieces, we were not able to fuse them due to limited time of our project. In the future, the lab hopes to fuse all three pieces and knockout TERT in Aspergillus nidulans to determine its localization.
Crystal Structure Analysis of Gallic Acid Decarboxylase and Pea Pathogenicity Protein
Nebojsa Markovic1, Cristina V. Iancu2, and Jun-Yong Choe2. 1Chemistry Major, Lake Forest College, Lake Forest, IL 60045; 2Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
Tannins are plant metabolites that play a role in plant defense. They are known for their binding and ability to precipitate proteins and thus are linked to different pathologies, including cancer. The hydrolysis of tannins produces Gallic Acids, which are used in the pharmaceutical industries for their anticancer and antioxidant properties. Gallic acids are difficult to degrade and often contribute to pollution. Gallic Acid Decarboxylase (GADC), a protein found in several different types of bacteria and yeast, is an enzyme that decarboxylates gallic acid to form pyrogallol, allowing the natural degradation of gallic acid. This research was conducted to determine the three-dimensional structure of GADC from Arxula adeninivorans (a type of yeast) and also of its homologue, Pea pathogenicity protein (PPP) form Madurella mycetomatis. For protein preparation, we cultured Escherichia coli cells containing the plasmid with GADC or PPP tagged with polyHistidine, and purified the proteins by immobilized metal affinity chromatography (IMAC). We crystallized the purified proteins and collected x-ray diffraction data on the protein crystals at Advanced Photon Source, Argonne National Laboratory. Crystal structures revealed the active site of the enzymes along with several metal binding sites, including cobalt and potassium sites. As expected, mutants of residues in the active site decreased or abolished enzyme activity. We are currently investigating the role of the metal sites in GADC and PPP.
Quantifying Functional Citrate Transport Protein from HEK293 Cells
Ronald Kaplan, June Mayor, Steven Stark and Tariq Aldaas. Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, North Chicago, IL, 60085
Plasma Membrane Citrate Transport (PMCT) is a protein found in H. sapiens, M. musculus, R. rattus, and C. elegans plasma membranes that allows cells to transport citrate from the circulatory system, leading to the production of fatty acid, triacylglycerol, and cholesterol. Citrate is produced in the mitochondria during the citric acid cycle and eventually leads to a yield of ATP through oxidative-phosphorylation. The PMCT has not been studied as much as other citrate transporters due to the difficulty of reproducing a favorable eukaryotic environment for the protein. Human Embryonic Kidney cells (HEK293) were used for the study due to past scientists’ successes in overexpressing membrane proteins from the cells. The goal of the experiments was to determine the best procedure for obtaining large quantities of functional citrate transport protein from HEK293 cells. The results would prove very important in later testing specific inhibitors that could limit the function of CTP in humans. The hypothesis was that reconstituting CTP in model liposomes would lead to functional CTP that would constitute a model system of citrate transport. Tests were run to determine the quantity of purified CTP and how the level of CTP function. The results indicated that CTP was overexpressed but the CTP was not functional. Future studies could be run to obtain functional CTP for testing inhibitors that could be used in manufacturing drugs for targeting CTP in humans.
Developing a Viral Vector For Neurodifferentiation and Retrograde Tracing
Schuyler Kogan, S. Schrank, R. Marr; Department of Biology and Neuroscience Program, Lake Forest College, IL 60045, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
As part of a larger study to investigate the roots of neurodegeneration in Alzheimer’s disease using induced neuron cultures as a model organism, it is necessary to assess the functional properties of these cells. In order to determine the extent that induced neurons can form functional synaptic connections, we aimed to develop a viral vector carrying genes capable of inducing neurodifferentiation, as well as tracing genes so that the cells’ connective properties can be measured using a rabies-based virus for retrograde labeling. Three initial goals to accomplish this task were to prepare a plasmid carrying the transcription factor Neurogenin2 (Ngn2) for the addition of tracing genes, to assess the strength of the CAG promoter, and to improve the CAG promoter by shortening the intron. We successfully prepared a vector based on the original Ngn2 plasmid and a DNA sequence with multiple restriction enzyme sites for the tracing genes to be implanted. We developed a shorter intron to replace the original sequence, although the promoter did not demonstrate as high expression as another promoter. We are currently in the process of constructing the new vector so that the connective function of induced neurons can be measured in culture and, eventually, in vivo.
UNDERSTANDING PARKINSON’S DISEASE IN YEAST MODELS: What α-synuclein’s New Familial Mutants Tell Us About Toxicity
Chisomo Mwale, Emily Ong, Maiwase Tembo, Charles Alvarado,Michael Buabeng, Natalie Kukulka, and Shubhik DebBurman. Department of Biology and Neuroscience Program, Lake Forest College, IL 60045
Parkinson’s disease (PD) is associated with the aggregation and misfolding of alpha-synuclein in midbrain dopaminergic neurons. The gene for alpha-synuclein has six known mutations that directly cause familial forms of PD. The pathological determinants of three of these mutants (A30P, E46K, and A53T) are well characterized in diverse model systems and they that reveal that each mutant affects cellular toxicity in distinctive ways. The three more recently discovered familial mutants (H50Q, G51D, and A53E) are not extensively studied. We expressed H50Q, G51D, and A53E mutants in budding and fission yeasts model systems and hypothesized that each would generate toxicity by altering their membrane association and aggregation properties, and by disrupting cellular pathways including nitrative stress responses and endocytosis, but each would do so in distinctive ways. First, we found that the H50Q and A53E mutants were toxic to yeast, and bound membranes and aggregated within yeast, while G51D was cytoplasmically diffuse and nontoxic. Secondly, we asked whether the loss of the original amino acid or the gain of the new amino acid in each new familial mutant is responsible for disease. We created four substitution mutations for H50Q, G51D, and A53E in both yeasts models corresponding to the four functional classes of amino acids. We found that H50D was cytoplasmically diffuse and nontoxic, G51A bound membranes and aggregated like WT, G51E was cytoplasmically diffuse and nontoxic like G51D, and A53R was cytoplasmically diffuse and nontoxic, suggesting both the loss of the original amino acid and the gain of the new amino acid are key. Thirdly, we found that the new mutants aggregated more and were more toxic in certain strains altered for increased nitrative stress. Collectively, this work adds insight into the pathogenicity of different familial PD mutants of alpha-synuclein.
The effect of phenytoin on respiratory rhythm and pattern generation
Emily Fink, and K. Kam, Department of Cell Biology, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
Phenytoin is an anti-epileptic drug that reduces seizures by blocking specific sodium channel subtypes, but also has sedating effects when administered. These same sodium conductances are important in the normal function of many neural circuits, including the medullary nucleus controlling breathing, the preBötzinger Complex, that may contribute to its sedating effects. To determine how phenytoin may be affecting other brain regions, we explored the effects of phenytoin on the neural control of breathing in rhythmic medullary slices from neonatal mice. Applying phenytoin to slices had varying effects on both the amplitude and frequency of rhythm. After the initial application of phenytoin, the interval between bursts increased, but then decreased with continued application. Application of phenytoin initially increased burst amplitude, but then decreased burst amplitude after a few minutes. Long applications of phenytoin reversibly blocked rhythmic activity. These effects were seen in both preBötzinger Complex and hypoglossal nerve, a physiologically relevant respiratory motor output. We conclude that phenytoin has a modest inhibitory effect on the rhythmic activity of the preBötzinger complex that may contribute to its sedating effect.
Quantification of Small-conductance Calcium-activated Potassium Channel SK2 Expression in the Rat Basolateral Amygdala
Phillip Freund, J. Vantrease, and J. A. Rosenkranz. Department of Biology, Lake Forest College, IL 60045; Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
The basolateral amygdala (BLA) is a brain region that has been shown to be important in the neural processing of stress and anxiety, and may be contribute to sex differences in anxiety disorders, as increased neuronal activity in the BLA has been connected to increased levels of anxiety. The small-conductance calcium-activated potassium channel SK2 protein is an important ion channel in central nervous system neurons, and regulates neuronal activity by decreasing and controlling the firing rate of neurons. Preliminary quantitative polymerase chain reaction (qPCR) data showed no sex difference in SK2 mRNA levels in the rat BLA, however, Western blot data showed higher levels of SK2 protein in male rats. There was no significant difference in SK2 mRNA or protein expression in the female rat population based on estrus (reproductive) cycle phase. This suggests SK2 expression differences arise after the SK2 protein is made, and are estrus cycle independent. Based on this data it was hypothesized that BLA SK2 protein expression would be higher in male rats. Before this hypothesis could be investigated, the immunohistochemistry (IHC) protocol (which visually displays SK2 protein expression and distribution in rat brain tissue) had to be optimized to ensure that it is reliable and accurate. To do this, the estrus cycle stage of the female rats was tracked using vaginal lavage. IHC staining for SK2 protein was conducted on rat brain tissue, and it was found that perfusing rats with glutaraldehyde (a fixing agent) led to better staining, as did higher concentrations of the IHC antibodies. These results will prove important when conducting IHC to quantify SK2 expression and distribution in the rat BLA.
The relationship between the nucleoporin complex, NUP88/214, with the viral latency protein, LANA2, in KSHV pathogenesis
Sabrina Najibi, Sharma-Walia, N. Microbiology and Immunology Department of RFUMS, North Chicago, IL 60064
KSHV (Kaposi’s sarcoma-associated herpesvirus) is a cancer virus affecting people affected with HIV and AIDS. Herpes viruses are part of three classes of viruses (alpha, beta and gamma) where KSHV is in the gamma class of herpes viruses. The Nucleoporin complex (NPC) which is comprised of roughly 30 proteins that are found at the nuclear envelope and are comprised of nucleoporins. Nucleoporins help transport proteins, mRNA, and even viral DNA such as KSHV in and out of the nucleus. In KSHV infected cells, NUP88 and NUP214 were studied as they form a complex together and are found at the cytoplasmic side of the NPC. In addition, LANA2, which is a latent protein found specifically in a type of cell known as Primary Effusion Cells (PEL) was studied. LANA2 is shown to be involved in maintaining the integrity and promotes tumorigenic growth in the cells as well as upregulating various inflammatory pathways which contribute to the pathogenesis of KSHV. In other words, this protein helps the cell grow abnormally. I predict that LANA2 interacts with NUP88 and NUP214 based on the recent primary literature viewed and I hypothesized that LANA2 becomes misplaced when Lipoxin, which is an anti-inflammatory drug used to treat KSHV infected cells. Several proteins that have been shown to pass through NUP88 and NUP214 have been expressed in cells infected with KSHV and those cells being treated by Lipoxin. Previous findings from this lab has shown the effects of Lipoxins as they have been seen to promote cell programmed death, as well as reducing the expression of several lytic genes, thus reducing cellular proliferation. In conclusion, I predict that LANA2 would be misplaced from NUP88 and NUP214 under the effects of Lipoxin.
Not All Part-Set Cues are Created Equally
Magdalen G. Kroeger, N.L. Hueng, S.D. Curry, M.B. Copeland, M. Kelley. Department of Psychology and Neuroscience program, Lake Forest College, IL. 60045
The present study manipulated the memorability (high vs. low) of the part-set cues provided to participants after they read a series of three short paragraphs, as well as whether cues were present or absent at test and whether the test required free or ordered recall. The paragraphs, idea units, and memorability data were drawn from Reysen et al. (2009)—the high memorability cues comprised the seven most memorable idea units for each paragraph, whereas the low memorability cues were the seven least memorable idea units. When the free and ordered data were scored using a lenient (free recall) criterion, part-set cueing inhibition only occurred when high memorability cues were presented; providing low memorability cues at test eliminated part-set cueing inhibition. With a relative order scoring criterion, the same pattern emerged, which was surprising because part-set cues tend to enhance order memory.
UNDERSTANDING PARKINSON’S WITH YEAST: How are α-Synuclein’s N- and C-domains relevant to disease?
Alexsandra Biel, Niam Abeysiriwardena, Yoan Ganev, Katrina Campbell, Michael Fiske, & Shubhik DebBurman. Biology Department and Neuroscience Program, Lake Forest College, Lake Forest, IL 60045
Parkinson’s disease (PD) is a neurodegenerative disorder linked to the loss of dopaminergic neurons in the midbrain. Consequently, symptoms such as muscle rigidity, tremors, and bradykinesia appear. A key pathological marker of PD is the presence of Lewy bodies, which are mainly composed of misfolded α-synuclein protein. α-synuclein is divided into three domains: N, M, and C domains. The six familial mutations implicated in PD are concentrated on the N-domain, which has been implicated in membrane binding. The C-domain is involved both in posttranslational modifications and naturally occurring truncations, and it is important to the solubility of the protein. First, using a yeast model system, we present evidence that all three domains of α-synuclein are needed to generate α-synuclein‘s membrane localization and organismal toxicity. Second, we show that naturally-occurring C-terminal truncations reduce the protein’s solubility and membrane association. Third, we show that the location of an attached tag changes the solubility of α-synuclein. The amino acids key to each domain’s function remain unknown, so we hypothesize that key parts of a domain (rather than the entire domain) underlie the functions ascribed to that domain. To identify the key regions that are important to membrane-binding and solubility, respectively, we modified α-synuclein by progressively truncating the protein at either the N- or C-terminal of the protein and we have expressed them in budding yeast. We have begun functional assays to test the properties of these truncations. In the future, we will conduct studies to further understand the physical nature of the protein.
INSIGHT INTO PARKINSON’S DISEASE FROM YEAST: Do familial mutations impact the disease contributions of a-synuclein’s covalent modifications?
Ariane Balaram, Joseph Mountain, Chisomo Mwale, Rosemary Thomas, Yoan Ganev, Alexandra Roman and Shubhik DebBurman. Biology Department and Neuroscience Program, Lake Forest College, Lake Forest, IL 60045
Parkinson’s disease (PD) is a neurodegenerative disorder linked to the loss of midbrain dopaminergic neurons. A key pathological feature, Lewy bodies, are aggregates composed mainly of misfolded protein α-synuclein. Six familial mutants (A30P, E46K, A53T, H50Q, G51D, A53E) of α-synuclein have been linked with early onset Parkinson’s disease. In addition, evidence shows that α-synuclein undergoes multiple post-translational modifications. One modification, sumoylation, takes place on lysine-96 and lysine-102 and, based on limited studies, is proposed to have neuroprotective effects by increasing α-synuclein solubility. In contrast, the modification of phosphorylation, mainly on serine-87 and serine-129, is still debated as evidence suggests both neuroprotective and neurotoxic properties. Past research in our lab has focused on the effects of sumoylation and phosphorylation on the wild type α-synuclein, but less is known about the effects of combining the familial mutants of α-synuclein with modifications of sumoylation and phosphorylation. We hypothesized that the combination of familial mutations with sumoylation-deficiency and phosphorylation enhancement would show the most toxicity; but the level of that toxicity would depend on the individual familial mutant. To explore this hypothesis, the overall goal of our Richter project was to create each of the six familial mutations on three different versions of α-synuclein template DNA: sumoylation-deficient (K96R/K102R), sumoylation- deficient and phosphorylation-deficient (K96R/K102R/S87A/S129A), and sumoylation-deficient and phosphorylation- mimic (K96R/K102R/S87D/S129D). We used PCR-based site-directed mutagenesis to create these mutations on the three α-synuclein-templates. Here, we share our results that we successfully made all 18 variants and expressed them in budding yeast. Functional studies are already underway to the functions of these new variants in yeast.
Insight into Parkinson’s Disease from Yeasts: The combinatorial impact of post-translational modifications and familial mutations on α-synuclein
Yoan Ganev, Chisomo Mwale, Joseph Mountain, Ariane Balaram, Rosemary Thomas, Alexandra Roman, Morgan Marshall, & Shubhik DebBurman. Neuroscience Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Parkinson’s disease (PD) is a neurodegenerative disorder linked to the loss of dopaminergic neurons in the midbrain. A key pathological marker of PD is the presence of Lewy bodies, which are mainly composed of misfolded α-synuclein protein. α-synuclein is a highly post-translationally modified protein. While phosphorylation and nitration of α-synuclein are well studied in the context of PD pathology, less is known about sumoylation, which is proposed to be neuroprotective based on limited studies. The majority of sumoylation takes place on the lysine-96 and lysine-102 sites of α-synuclein, and it increases the protein’s solubility. The goal of this research was to better understand the role of sumoylation in regulating α-synuclein toxicity, and we performed four studies towards it. First, we evaluated the effects of blocking sumoylation on α-synuclein in the well-established budding yeast model for PD and found that α-synuclein becomes more aggregated, gains toxicity, and loses localization at the plasma membrane. Second, we evaluated the effects of altering sumoylation pathways by using yeast strains with reduced (ulp1ts) or excessive sumoylation (smt3ts), and found that α-synuclein aggregates more with reduced sumoylation, but becomes less toxic with increased sumoylation. Third, we asked how altering phosphorylation of α-synuclein would alter sumoylation’s protective role and found that blocking phosphorylation reduced the protein’s toxicity. Finally, we began evaluating whether blocking sumoylation and altering phosphorylation on familial PD mutant versions of α-synuclein would exacerbate its toxicity. In the future, we will conduct further studies to understand how sumoylation affects other variants and modifications of α-synuclein.
Alone, anxious, and unable to forget: Fear-potentiated startle (FPS) model of chronic social isolation in male rats
Michael Janeček and J. Dabrowska. Department of Neuroscience/Cellular and Molecular Pharmacology, Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064, USA
Environmental stressors are known to modify neuronal activity. Human studies suggest that social isolation induces anxiety and prolongs recovery from psychological trauma. Conversely, having strong social support enhances stress-coping strategies and reduces the risk of developing anxiety, generalized anxiety disorder (GAD), and post-traumatic stress disorder (PTSD). While most studies tackling isolation in rats use freezing to measure fear, the translation of such findings to humans remains controversial. Here, we instead utilize the fear-potentiated startle (FPS) paradigm, a valid measure of anxiety that is observable in both rodents and humans. In rodents, FPS consists of a 95dB white noise burst presentation (unconditioned stimulus, US/CS-) that elicits a whole-body startle reflex, quantifiable as force displacement. Yet, social isolation in terms of FPS is poorly understood: a PubMed search of “fear-potentiated startle” and “isolation” in “rat” yielded mere three results. To close this gap in knowledge, our objective was to investigate the effects of social isolation on cued fear extinction learning and non-cued fear expression. To this end, P40-P44 adolescent rats (N = 44) were housed singly for up to 22 days with no haptic and limited olfactory contact with conspecifics, while control animals were housed socially in pairs or trios. We report that animals isolated for 6 days had an enhanced cued fear expression and required more extinction trials to abolish their fear response to a visual cue (conditioned stimulus, CS+), compared to socially housed animals. Additionally, all animals were successfully fear-conditioned during a re-instatement session involving two shock presentations, whereupon isolated rats exhibited elevated non-cued fear. This finding suggests that fear memory re-instatement in individuals lacking social support may play a role in states of hypervigilance, marked by diminished threat-safety discrimination. Interestingly, 96-hour isolation prior to fear-conditioning did not affect the acquisition of fear memory. Although future research should replicate these findings, explore social isolation in females, and identify neural mechanisms underlying social isolation, this study presents a viable FPS model of social isolation in male rats.
Image-based classification of body language poses to emotions using convolutional neural networks
Aasimah Tanveer and Sugata Banerji, Department of Math and Computer Science, Lake Forest College, IL
Recognizing human emotions based on a person’s body language is a complex neurological process that we often take for granted. The visual pathway that propagates from the eye to the occipital lobe, breaking down images from basic features and building upon the previous layer, can be replicated in a computer using artificially intelligent algorithms in the field of computer vision using convolutional neural networks. The neural network, just like in the brain, models the interaction between neurons with each neuron, called a perceptron, being represented by a mathematical function. Real-world images of humans displaying emotion through body language of varying environments, colorings, and features are classified and used to train the neural network in the computer. This allows the algorithm to pick out features that occur in each emotion that will intelligently aid in classifying body language poses that it has never seen before to an emotion. The accuracy of the classification is then analyzed by reviewing what features were extracted from the images and the network is retrained accordingly to output even more accurate results.
Coordination of Insulin Resistance in Response to a High-Sugar Diet in Drosophila
Jeanne M. C. McDonald, Alexander W. Shingleton. Department of Biology, Lake Forest College, Lake Forest, IL 60045
While the consequences of diet-induced insulin resistance, such as type 2 diabetes, have been well described in humans, the molecular-genetic mechanisms underlying this condition are not yet well understood. Previous studies have shown that elevated levels of insulin-binding proteins are associated with diet-induced insulin resistance in humans, and other studies have shown that, in Drosophila, insulin resistance may be coordinated systemically by the fat body, the equivalent of the human liver. Therefore, one hypothesis is that high sugar diet (HSD) stimulates the release of insulin-binding proteins by the fat body, which downregulate insulin-signaling by inhibiting the activity of insulin-like peptides. We therefore tested whether downregulation of insulin-signaling on HSD is facilitated by the insulin-binding peptide ImpL2 using RNA interference (RNAi) to silence ImpL2 in the fat body via the gal4-UAS system. The flies were then reared across a range of diets increasing in sucrose concentration. By measuring body size and developmental time, we were able to demonstrate that HSD downregulated insulin-signaling in the control, as Drosophila reared on HSD were significantly smaller and developmentally delayed. However, when ImpL2 was silenced in the fat body, HSD had no effect on size or developmental time, indicating that insulin-signaling was not reduced in these flies. Therefore, these results suggest that insulin resistance is coordinated systemically by insulin-binding protein in response to high sugar.
Localization and Trafficking of the volatage-gated calcium channel UNC-2 inCaenorhabditis elegans
Kendra Fobert, K.H. Oh, H. Kim. Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064
UNC-2 is a protein-coding gene present in Caenorhabditis elegans which relates to a variety of functions, including the regulation of serotonin levels and normal movement control. The gene unc-2 encodes a channel-forming subunit of the voltage-gated calcium channel homologous to a P/Q type calcium channel found in humans. The improper function of calcium ion channels has been linked to a variety of nervous system disorders, signifying them for study and experimentation as possible targets for treatment. Calcium channels may be formed by a variety of combinations of several subunits, so a wide variety of these channels with a range of functions exist. The differences between specific channels are important to consider for use with drug therapeutics targeting calcium channels. Because of the diversity of calcium channels, there is still much to be learned about their form and function. In this project, C. elegans was used as a model organism to identify genes that control the localization and trafficking of unc-2 the voltage-gated ion channel. We took advantage of the transgenic line designed to express GFP-marked UNC-2. This transgenic line was put through mutagenesis and examined to find mutants with apparent reduction or loss of the UNC-2 level. In some mutants, unc-2 did not appear at all in the brain under fluorescence microscope, suggesting that the gene was knocked out completely. The motor capabilities of these worms were severely impacted, causing them to move at much slower speeds than worms of their original line. In a few other mutants, GFP levels are significantly reduced, suggesting that mutations may reduce UNC-2 levels by influencing UNC-2 stability or trafficking. It is possible that knocking out this gene accompanied other mutations which prevented the worms from reproducing successfully, blocking further study of those strains. These results are in alignment with the primary expression of unc-2 being located in motor neurons and 2 types of neurons related to egg laying. Further study must be done with these mutants to determine their specific genetic properties and exactly how their altered traits are related to calcium channel function.
FIYS106 Medical Mysteries of the Mind
Medical Mysteries of Language
Alexandra Dejneka, Leslie Gonzales, Courtney Levitt, Peter Simmeth, Madison Strejc, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how we communicate through language and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.
Medical Mysteries of Sex and Sexuality
Corynn Christjansen, Todd Fugelseth, Julius Khamoo, Grace Monshausen, Hasibullah Sadat, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying gender differences, identity and sexual preferences and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.
Medical Mysteries of Memory
Mackenzie Blythe, Olivia Drevs, Grace Gould, Bari Mohammed, Erica Saldana, Madeline Webb, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how learn and form memories and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.
Medical Mysteries of Thought
Samantha Cosmo, Kayenath Khan, Anthony Sullivan, Olivia Smith, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Students of FIYS106 Medical Mysteries of the Mind will present he neuroscience underlying how engage in thought processes and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.
Medical Mysteries of Sleep
Blakely Drake, Gina Dudek, Erick Quiroga, Estella Tcaturian, Jenna Wolfrum, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how we sleep and stay awake and they will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.
Medical Mysteries of Emotions
Tianna Beilke, Allison Fair, Hannah Gurholt, Dana Midani, Molly Schul, Sarah Truxton, First-Year Studies Program and Biology Department, Lake Forest College, Lake Forest, IL 60045
Students of FIYS106 Medical Mysteries of the Mind will present the neuroscience underlying how we express emotions and why we do it and will delve into several current medical mysteries that are the cutting of scientific research. They will educate the audience using posters, models and interactive activities.