Kenneth Weik Research Symposium Abstracts

Honoring Outstanding Student Research in Biology

These abstracts represent the hard work of students who will present at the Second Annual Symposium to be held on November 9, 2011, in the Skybox

2011 Symposium Abstracts

Ahlstrand

Ms. Kayla Ahlstrand ’12, Chicago, Illinois, Biology major

Faculty Sponsor: Dr. Matthew Kelly (Lake Forest College)

1.  Collaborative Inhibition Counteracts the Mnemonic Benefit of Social Information

In one experiment, participants read target words that were presented in the context of a social sentence ‘Willow towered over Meadow’ or a nonsocial sentence ‘The willow towered over the meadow.’  Subsequently, they received a surprise cued recall test for the targets nouns/names and completed the test either alone or in a group of two.  Despite the fact that the stimulus materials were held constant across conditions, participants showed a social processing advantage in memory—that is, they remembered the social (name) versions of the target words significantly better than the nonsocial (noun) versions.  Further, the social benefit was not strong enough to counteract the inhibitory effects of collaboration as collaborative groups recalled significantly fewer words than did nominal groups.  The present study also demonstrated robust collaborative inhibition with cued recall, a task previously assumed to eliminate such inhibition.

 

Bailey

Ms. Lauren Bailey ’12, Chattanooga, Tennessee, Biology major

Faculty Sponsor: Dr. Andrew Wilson (Chicago Botanic Gardens)
 

2. Sabrobic Fungi Associated with the Leaf Litter of Rhamnus cathartic

Leaves of the invasive European buckthorn, Rhamnus cathartica, rapidly decompose. As a result, buckthorn-invaded areas typically have bare or exposed mineral soils with sparse litter layers. The cause of such rapid litter decomposition is unclear but is thought to be due to the high nitrogen content of R. cathartica foliage stimulating and supporting saprobic microbial populations.  The contrast between buckthorn soils and that of organic humus soils in native stands of Quercus alba likely results in a shift of the microbial community composition.  This could play a vital role in the successful invasion, or lack thereof, by invasive exotics like R. cathartica. Because fungi are one of the primary decomposers of leaf litter in forest habitats, shifts in metabolic processes are likely reflected in the composition of leaf litter fungal communities. We explored this hypothesis, using molecular tools to describe the fungal communities associated with R. cathartica  leaf litter in its natural habitat. Using a combination of PCR, cloning, RFLP analysis and Sanger sequencing, we identified approximately 33 fungal OTUs associated with R. cathartica  leaf litter, which were all identified as ascomycete fungi. Next, these techniques will be applied to the leaf litter of the native Q. alba for comparison.

Buck Leong Ms. Jody Buck ’13, Seattle, Wisconsin, Neuroscience and English Major

Ms. Lani Leong ’14, Northfield, Illinois, Neuroscience Major

Faculty Sponsor: Dr. William Frost (Rosalind Franklin University of Medicine & Science)

3. Behavioral study of Aplysia locomotion and subsequent recovery of escape locomotion following surgical nerve crush

The marine gastropod Aplysia californica  is an ideal organism for neurophysiological research because of its large, easily accessible ganglion.  Aplysia also exhibits two types of locomotion: normal locomotion (crawling) and escape locomotion (galloping), which have not been well studied or defined in past literature. In this study, these two types of locomotion were qualitatively and quantitatively measured for comparative purposes. It was observed that escape locomotion is a faster and more exaggerated form of the crawl, and can be reliably induced. After behavioral experimentation was conducted,  we then replicated a study that involved surgically crushing the cerebropleural (C-Pl) connective in order to eliminate escape locomotion while still leaving crawling in tact (Jahan-Parwar & Fredman).  Escape locomotion was recovered post nerve crush indicating the ability of nerve fibers to recover and reestablish connections.

We had intended to build upon this study by adding an optical recording component to visualize how neural networks in Aplysia  work during functional recovery, however we were unable to attain accurate and reliable recordings from the experimental animals.

 

Campbell

Ms. Katrina Campbell ’14, Akron, Colorado, Neuroscience and Biology major

Faculty Sponsor: Dr. Shubhik DebBurman (Lake Forest College)


 

 

 

4. Identifying Specific Amino Acids in Parkinson’s Disease Protein α-synuclein that Control its Toxic Properties

Parkinson’s disease (PD) is a common neurodegenerative disease in which neurons in a specific region of the midbrain die. This loss results in a variety of symptoms including tremors, rigidity, and other motor and cognitive problems. A PD hallmark is the aggregation of a membrane-associated protein called alpha-synuclein in these dying neurons. Proteins are made of amino acids, and while each amino acid contributes to the overall protein structure and function, certain amino acids are typically more important than others. My research project involves testing, in yeast models, a subset of five specific amino acids recently implicated to be important in alpha-synuclein’s PD-pathology related properties: namely, its ability to aggregate and associate with membrane phospholipids. First, single points were created where each of the four amino acids (D2, A76, V77, and E83) were mutated to a different amino acid to either enhance (E83A, E83V) or eliminate (D2A, A76E, V77E) their predicted contributions to the above two pathological characteristics. Then, we assessed each mutant’s ability to alter alpha-synuclein 1) association with phospholipid membranes, 2) aggregation, and 3) toxicity to yeast cells. Our preliminary data for each of these five amino acids supports their relevance in alpha-synuclein’s contributions to PD pathology, strengthening previous predictions in the field and the usefulness of yeasts as model organisms for studying molecular mechanisms underlying the spectrum of protein misfolding diseases. 

Cole

Ms. Sydni Cole ’12, Northbrook, Illinois, Neuroscience major

 Faculty Sponsor: Dr. Matthew Kelley (Lake Forest College)

 

5. Part Set Cuing Effects in Object-Location Memory

Part-set cuing refers to the presentation of cues during recall tasks.  Previous studies have shown both inhibitory and facilitative effects of cues on recall of the remaining items.  The present study was designed to test the part-set cuing effects of object-location memory using snap circuits as stimuli.  Participants watched a video of a 16-piece (8 pieces and 8 connectors) snap circuit assembly and then were asked to reproduce the finished “object.”  Participants were either cued or uncued, with the cues consisting of 4 of the connections between pieces.  Data was then recorded as to the number of correctly placed pieces as well as the number of correct connections between pieces.  Results indicate no significant difference in performance between the cued and uncued participants, which is consistent with previous studies done using chess positions.

 

Gilbow

Mr. Dillon Gilbow ’13, Blytheville, Arkansas, Chemistry major

Faculty Sponsor: Dr. Ronald Kaplan (Rosalind Franklin University Medical School)


 

 

6. Functionality of PMCT in Proteoliposomes

Citrate is an integral intermediary of the TCA cycle and serves as a substrate for biosynthesis of many lipids.   Citrate is either produced in the mitochondria during the TCA cycle or obtained from the extracellular environment.  In order for citrate to be transported into the cell and out of the mitochondria it must pass through cellular membranes.  In the mitochondria citrate is transported out via the Citrate Transport Protein (CTP) and Plasma Membrane Citrate Transport Proteins (PMCT) transports citrate from extracellular environments into the cell.   Our research has recently been focused on studying PMCT and how ionophores such as Valinomycin and CCCP affect PMCT’s functionality.   Through transport kinetics studies our current findings have shown that ion and proton concentrations have a major effect on PMCT’s ability to transport citrate across the plasma membrane.  A better understanding of how to inhibit citrate transport could mimic a caloric restriction diet regiment by restricting lipid production creating possible therapies in obesity and diabetes.

Kuprys Hauer

Mr. Tyler Hauer ’13, Toronto, Ontario, Biology major

Mr. Paulius Kuprys ’12, Riverside, Illinois, Biology major

Faculty Sponsor: Dr. Karen Kirk (Lake Forest College)

7. Identifying the First Telomerase RNA in a Filamentous Fungus

The ends of eukaryotic chromosomes are protected by nucleoprotein structures termed telomeres, which are synthesized by the essential enzyme, telomerase. We are studying the telomeric components of the Aspergilli, as these filamentous fungi have exceptionally short telomeres. A key component of telomerase, the telomerase RNA, has not been identified inany filamentous fungus. The telomerase RNAs from even closely related organisms are highly divergent and thus are difficult to identify through bioinformatics. Since Aspergillus oryzae  has a particularly long telomeric repeat of 5’ TTAGGGTCAACA 3’, and we knew the telomerase RNA must contain this sequence as a template to synthesize the telomeres, we were able to identify roughly 20 candidate sequences in the A. oryzae  genome. Experiments utilizing RTPCR and PCR have shown that one candidate located on chromosome III is transcribed whereas all other candidates in the genome were not transcribed. The 5’ end of this RNA sequence has been identified using RLM-RACE to be at most 301 bp from the start of the template, but the 3’ end remains undetermined. Successive experiments using 3’ RACE have proved inconclusive indicating that the 3’ end of the telomerase RNA could lack a poly(A) tail. Using RT-PCR, we determined that the 3’ end is between 1567 and 1775 bp from the template. These results suggest that the total telomerase RNA length in A. oryzae  is between 1868 and 2076 bp. The length is plausible as it is similar to the telomerase RNA lengths of yeasts. This suggests that we may have discovered the first telomerase RNA in a filamentous fungus.

Johnson

Ms. Kristi Johnson ’12, Sheridan, Illinois, Neuroscience major        

 Faculty Sponsor: Dr. Amiel Rosenkranz (Rosalind Franklin University of Medicine & Science)

8. Does Chronic Stress Cause Changes in Emotion-Related Behavior via Activation of the Immune System

Stress is an expected component of everyday life. However, chronic stress can cause a range of impairments, including exacerbation of psychiatric disorders. In particular, stress has a negative impact in patients with depression. Chronic stress and depression are both associated with hyperactivation of the amygdala, a brain region involved in mood and emotion. This commonality has led to the suggestion that chronic stress induces depression by causing a hyperactivity of the amygdala. However, the pathway from chronic stress to hyperactivity of the amygdala is unknown. Evidence has emerged that implicates the immune system in some of the detrimental effects of stress on affective behavior. Here we test the hypothesis that chronic stress causes a change in amygdala function through activation of the immune system. To test this hypothesis, we subject adult male rats to a repeated stressor over the course of 10 days, and measure immune factors in the plasma, behaviors that depend upon the amygdala, and activation of the amygdala. In a separate set of experiments, we test whether suppression of the immune response to stress alleviates the impact of stress on these measures. Preliminary results suggest that chronic stress causes activation of the amygdala and increased amygdala-dependent behaviors. Determination of the role of the immune system in the detrimental effects of stress may uncover new insights into how stress can impact psychiatric disorders, and potentially lead to novel therapeutic options to treat these effects of stress.

Rida Khan

Ms. Rida Khan ’14, Gilberts, Illinois, Biology and Chemistry major

Faculty Sponsor: Dr. Shubhik DebBurman (Lake Forest College)

 

9. Developing assays to evaluate cross regulation between multiple degradation pathways for Parkinson’s disease protein Alpha-Synuclein

Most neurodegenerative diseases are characterized by the accumulation of a specific protein. For Parkinson’s disease (PD), the culprit is alpha-synuclein. Therefore, the ability to breakdown the buildup of alpha-synuclein is of prime therapeutic interest and for drug design the molecular basis of such breakdown must be first understood. Previously, others and we have shown that both major protein degradation organelles found in our cells (the lysosome and the proteasome) are breakdown sites for alpha-synuclein. Whether these two organelles act independently to degrade alpha-synuclein or cross-regulate each other is not known. We support the latter notion, and therefore, predict that compromising both pathways will combine to accelerate alpha-synuclein pathology. To test this hypothesis, we need to develop effective ways to inhibit both pathways minimal cellular damage. For my Richter project, my first goal was to develop an assay to chemically inhibit the proteasome in a yeast model and then evaluate how such inhibition would alter the overall pathological properties  of alpha-synuclein. Specifically, I evaluated if two PD-related α-synuclein characteristics (sub cellular localization, and accumulation) are altered or worsened in yeasts treated with a known proteasomal inhibitor, MG132. My preliminary data suggests that while yeasts treated with MG132 relocalize alpha-synuclein into cytoplasmic aggregates, the protein accumulates less in these cells. I will present my experimental design and share this preliminary data.

Sohail Khan

Mr. Sohail Khan ’14, Lindenhurst, Illinois, Computer Science major

Faculty Sponsor: Dr. Dmitri Boudko (Rosalind Franklin University of Medicine & Science)

 

10.  Cloning of the Snf2 Gene of C. elegans

The Snf 2 gene in C. elegans  expresses a nutrient amino acid transporter (NAT), which is present not only in the roundworms but also various species of mosquito.  In this study, the gene was cloned via the use of ligation into various vectors, transformation into E. coli  cells, amplification using polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE), and finally extraction of the plasmid from cultures of the transformed bacterial cells.  Once the gene was cloned and sequenced to make sure it was the correct gene, it could be sub-cloned into expression vectors, such as the xenopus oocyte, and used to examine the function of this protein which could lead to discoveries in controlling roundworm and mosquito population, both of which have a vital role in many diseases.

Billy Levinson and Hillary Wind

Mr. Billy Levinson ’13, Northbrook, Illinois, Biology major

Ms. Hilary Wind ’14, Oak Lawn, Illinois, Biology and Environmental Studies major

 Faculty Sponsor: Dr. Anne Houde (Lake Forest College)

 

 

 

11. The effects of familiarity of females to males on male behavior in the guppy, Poecilia reticulata

 Guppies serve as a model organism for the study of sexual selection. Previous studies have shown that females prefer novel males. We wondered how this female preference for novel over familiar males in turn affected male behavior. This is because familiar males may be less successful at attracting females than unfamiliar males. We predicted that we should see consistent differences in behavior (through the number of courtship displays and gonopodial thrusts) when comparing males interacting with females who find them novel versus familiar, and also between days of the treatment cycle. This study is the first time a student has examined male success versus failure over time. Females showed a higher responsiveness to novel males for the majority of days measured. Male displays and thrusts differed between familiar and unfamiliar groups and depending on whether receptive females were present, but not in quite the pattern we predicted. Results are still in the preliminary stages of analysis, and more statistics need to be run.

Pinto

Ms. Samantha Pinto ’13, Toronto, Ontario, Biology & Studio Art Major

Faculty Sponsor: Dr. Dominik Duelli (Rosalind Franklin University of Medicine and Science)

12. Identifying microvesicles in metastatic breast cancer cell lines using homemade antibodies

 Breast cancer is widely known to be one of the deadliest cancers in the world. Annually, 1 in 8 or 12% of women are diagnosed with breast cancer, in the United States alone. Tumors originate from the inner lining of the milk ducts or the lobules that supply the ducts with milk.  Previous studies have led to the suggestion that micro RNA’s in body fluids could be designated biomarkers of disease. Previous research has als  o found miRNAs to be selective in their extracellular as well as intracellular release. MicroRNAs have been found to be associated with microvesicles (spheres, exosomes, and exovesicles). Studies have shown that microvesicles are released from breast cancer cells, where they migrate to other organs in the body creating a metastatic cancer cell line. In order to find the miRNA’s that trigger breast cancer, the first step is to look at the microvesicles and define their protein profiles. The Western Dot Blot approach was used to identify numerous vesicles containing different proteins. It was achieved using homemade antibodies to test which of them were more likely to bind to particles released from breast cancer cell lines. We find that although there are similarities in peaks among different cell lines, there also happen to be differences. These findings propose the idea that there are various unknown proteins and microvesicles released into body fluids that target specific organs in the body, metastasizing the cancer.

 


Senagolage

Ms. Madhavi Senagolage ’12, Colombo, Sri Lanka, Biology major

Faculty Sponsor: Dr. Shubhik DebBurman (Lake Forest College)

 

 

13. Complex Regulation of α-synuclein Properties in Yeasts by Endocytosis Pathway Genes

Parkinson’s disease is an incurable and neurodegenerative disease linked to the accumulation of α -synuclein protein. The impaired degradation of α –synuclein is thought to promote its aggregation and cell death. Therefore, accelerating the degradation of α-synuclein is of therapeutic interest. We hypothesized that a-synclein uses MVB pathway as a route to the lysosome for degradation. Specifically, we evaluated if three disease-related properties (subcellular localization, accumulation and cellular toxicity) of α -synuclein changed in yeast strains when genes were eliminated that encode for seventeen proteins that control the endocytosis pathway. Our data suggested that most gene deletions altered at least one of the above properties related to pathology of α -synuclein. Therefore, in both wildtype and familial mutant E46K α -synuclein is degraded by the lysosome via the endocytosis route.

 


Sullivan

Mr. Peter Sullivan ’12, Lockport, Illinois, Biology and Chemistry  major

Faculty Sponsor: Dr. Anthony West (Rosalind Franklin University of Medicine & Science)

 

 

14. Inhibition of PDE10A modulates L-DOPA-induced dyskinesias in a dose-dependent manner in Parkinsonian rats.

Cyclic nucleotide phosphodiesterases (PDEs) regulate neuronal activity by regulating cAMP/cGMP signaling within the striatum. Recent studies indicate that PDE inhibitors could  attenuate L-DOPA-induced dyskinesias in Parkinson’s disease patients and that PDE10A, specifically, is a potential target for the treatment of hyperdopaminergic neuropsychiatric disorders. This study assessed the ability of the PDE10A inhibitor TP-10 for reversing behavioral correlates of L-DOPA induced dyskinesias in unilateral 6-OHDA-lesioned rats. Animals received vehicle, L-DOPA/ benserazide, L-DOPA/ benserazide with TP-10 (0.32 mg/kg), or L-DOPA/ benserazide with TP-10 (3.2 mg/kg). Rats were evaluated for frequency and intensity of abnormal involuntary movements (AIMs) 40 minutes after treatment administration. L-DOPA co-administration with a high, but not a low dose of TP-10, significantly attenuated total L-DOPA-induced AIMs as well as individual L-DOPA-induced AIMs such as left forepaw dystonia and hyperkinesias. Interestingly, the low dose of TP-10 decreased onset latency and increased intensity of L-DOPA-induced dystonia. Importantly, normal behaviors were not significantly affected across lesioned groups. These observations demonstrate that robust PDE10A inhibition reduces the incidence and severity of AIMs observed in dyskinetic rats and indicate that these effects are not due to decreased behavioral activation. Thus, selective targeting of PDE10A signaling may symptoms of L-DOPA-induced dyskinesias in patients with PD.

Szutenbach


Ms. Anneliese Szutenbach ’12, Chicago, Illinois, Biology major

Faculty Sponsor: Dr. Pliny Smith (Lake Forest College)

15.   The Search for M138 Mutant Alleles in the Model Organism: C. elegans

Caenorhabditis elegans, a microscopic hermaphroditic nematode with only 959 somatic cells, serves as a great biological model to study development. C. elegans develop and reproduce rapidly, are transparent, and each cell can be traced in its defined patterns of lineage. It has therefore been used extensively in studies of conserved genetic pathways.  Our current research includes RNAi experiments as well as genetic mapping of the mutant line of C. elegans, M138, in attempt to find the mutant alleles.  The strain exhibits an abnormal, nonfunctional pharynx, with differentiation of the muscle cells causing severe disorganization. RNA interference works to silence genes by introducing homologous double stranded RNA to specifically target a gene’s product.  It enables us to induce suppression of the highly specific genes in interest, which will give great insight in determining the gene(s) responsible for the M138 mutation.  The discoveries in C. elegans research provide a pathway into a better understanding of the full range of genetics in multicellular eukaryotes.

Vlaar

Mr. Ryan Vlaar ’14, Vale, Colorado, Environmental Studies major

Faculty Sponsor: Dr. Shubhik DebBurman (Lake Forest College)


 

16. Examining different mutations in α-synuclein leading to cellullar toxicity

Genetic mutations cause 10% of Parkinson’s disease (PD) cases. The best-studied gene is α-synuclein, and its three PD-causing mutations (A30P, E46K, and A53T)  that alter three different amino acids (the protein’s building blocks). Each mutant affects α-synuclein’s properties in a distinct way, indicating each building block’s strategic importance. Our hypothesis was that combining multiple mutations would cause a blend of different α-synuclein’s phenotypes, leading to higher cell toxicity. We evaluated these combinational effects in Fission and Budding Yeasts, two model organisms for Parkinson’s disease developed in the DebBurman lab. Using these yeast models allowed us to analyze differences in how α-synuclein localizes, accumulates, and alters cell growth. Specifically, we compared the properties of A30P/E46K, A30P/A53T, E46K/A53T, and A30P/E46K/A53T to the properties of single mutants and wild type genotypes of α-synuclein. Our primary hypothesis was not supported, as our data showed the phenotypic dominance of one mutation (A30P) over the two other mutants in both yeast models. We also found that a combination of mutations did not necessarily cause an increase in toxicity in the cell, at least in budding yeast.