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Undergraduate Research

zebra
Neuroscience majors design and conduct original research projects mentored by faculty members. These research experiences build critical thinking skills in addition to honing laboratory skills. Students then have the opportunity to present their findings at regional or national science meetings. Previous student work has employed two popular model organisms, C. elegans (a nematode), and  D. rerio (zebrafish), and has utilized software and hardware (video tracking) developed by computer science students to monitor animal behavior in different pharmacological exposures, mutational and phenotypic backgrounds, and learning and memory paradigms.

These Neuroscience undergraduate research projects were presented at the 2016 Belmont Science Undergraduate Research Symposium (SURS):

“The Relationship Between Acute Stress and Behavioral Sensitization in Apomorphine Treated Danio rerio

John Longenecker
Faculty Advisor: Lori McGrew, PhD

Zebrafish are a model organism for neuroscience research because their central nervous system is similar to humans’. One such similarity are dopamine receptors, a class of GPCRs. Dopamine is the primary neurotransmitter involved in the reward pathway. Psychomotor stimulants such as cocaine and amphetamines act on dopamine pathways. Furthermore, after repeated exposure to such drugs, the motor stimulus response is enhanced, a principle called behavioral sensitization. To combat this occurrence, some rehabilitation clinics help to gently lower patients’ dopamine levels by prescribing other dopamine agonists. One such drug is Apomorphine, which is only recently being investigated in this area. The hypothesis of this study was that apomorphine will increase dopamine levels and elicit anxiogenic behavior, which is another side effect seen during rehabilitation. Data was analyzed using a two-factor ANOVA, with replication. Results indicate that Apomorphine does not elicit anxiogenic behavior, which may add to a growing body of evidence that suggests Apomorphine is a suitable drug for rehabilitation clinics.

“Chemotaxis of Caenorhabditis elegans with Bacillus thuringiensis 4A4 in vegetative and sporulated growth stages”

Meghan McGath
Faculty Advisor: Robert Grammer, Ph.D.

A study performed by Angel Brothers at Belmont University looked at the attractant or repellent properties of vegetative B. thuringiensis 4A4 and fifth, sixth, and seventh day sporulated B. thuringiensis 4A4. The study observed what occurred to the C. elegans during the first and seventh hour of chemotaxis for the vegetative state and the first and twenty-fourth hour for the sporulated state. The goal of the current study is to find out what occurs to the C. elegans during the first, fourth, and tenth hours of a chemotaxis assay with vegetative B. thuringiensis 4A4 and the fourth, tenth, and twenty-fourth hours on the fifth, seventh, and ninth days for sporulated B. thuringiensis 4A4. Results point to the C. elegans being attracted to the vegetative B. thuringiensis 4A4 and not the control, water. Likewise, that are attracted during the beginning of chemotaxis for the sporulated B. thuringiensis 4A4. However, at the twenty fourth hour it was shown that the C. elegans began to chemotaxis towards the control, water. Findings indicate that C. elegans followed the predicted hypothesis of moving towards the B. thuringiensis 4A4.

“Investigation of p53 activation in HCT116 with the sesquiterpene Beta-Caryophyllene”

Diana Neculcea
Faculty Advisor:  Chris Barton, Ph.D.

Beta-Caryophyllene is a natural bicyclic sesquiterpene widely found in the essential oils of many common herbs and plants. The biologic properties of B-Caryophyllene have experimentally demonstrated anti-proliferative, antioxidant, antimicrobial, and anti-inflammatory effects. In this experiment, we investigated variances in the anti-proliferative effects of B-Caryophyllene on the isogenic human colorectal cancer cell line HCT116 with wild-type p53 (+/+) and HCT116 without p53 (-/-). Both conditions showed a dose-dependent response in cell growth inhibition to treatment with increasing concentrations of B-Caryophyllene. The nontoxic dose of 150uM B-Caryophyllene significantly decreased the mitotic cell count in the p53 (+/+) HCT116 by over 50% in the first 24-hours. The same IC50 value was used with the p53 (-/-) cells for consistency, however there was only a 10% decrease in the first 24 hours. This discovery suggests that the presence of p53 may play a role in the effectiveness of B-Caryophyllene as an anticancer agent.  

“The Effect of Anandamide on Working Memory in Danio rerio

Sam Zacovic
Faculty Advisor: Lori McGrew, Ph.D.

Anandamide is a naturally occurring psychoactive cannabinoid that neurons release to inhibit the stressful effects of cortisol. Anandamide interacts with CB1 and CB2 receptors, located in the central and peripheral nervous systems. This endocannabinoid system is active in a wide range of physiological processes such as sleep, hunger, memory, and stress. CB1 and CB2 receptors are present throughout the medial pallium in the amygdala and in the hippocampus, structures that are thought to be involved in associative learning. Zebrafish (Danio rerio) were chosen for this study because of their extensive endocabinnoid system. The interaction of cannabinoid activation with working memory was examined. The fish executed a T-maze task with reward and punishment training. The right arm of the maze contained a reward (conspecifics) while the left arm of the maze included a punishment (confinement). The fish were trained until they could consistently navigate to the right arm of the maze. Once trained, the fish were submerged in anandamide solution to stimulate internal cannabinoid activity. It was hypothesized that a lower dose of anandamide would not have an effect on T-maze performance, but a high dose would produce inhibitory effects on their recently learned association.

“The Effects of Dopamine on Learning and Certainty in Zebrafish”

Stephane Morin
Faculty Advisor: Lori McGrew, Ph.D.

Previous studies suggest that the neurotransmitter dopamine plays a crucial role in the development of memories and learning via positive reinforcement. Dopamine has been shown to influence aversion learning as well. (Schmidt, R., G. Morris, E. H. Hagen, R. J. Sullivan, P. Hammerstein, and R. Kempter, 2009). In a study done early this year, it was shown that situations in which animals learn to avoid negative stimuli are regulated by dopamine (Dani, John, PhD., 2016). In the present study, this effect was explored to determine whether extra dopamine could further increase the natural learning ability of Zebra fish. It was hypothesized that the increase in dopamine in the fish would lead to a greater learned response. To test this, the fish were all trained in the same T-maze under the same conditions. The fish were split into three groups – a control group – a pre dose group - a post group. The maze testing followed a basic reward - punishment system to activate both positive reinforcement and aversion pathways. After the final testing for each group, data were analyzed to determine whether each fish took the right path on the T-maze; the total time spent to complete the maze; and the time each fish spent latent in the T-maze. These figures can help determine whether the fish knew the right choice, and how certain the fish were in that decision (time latent compared to total time).


These Neuroscience undergraduate research projects were presented at the 2014 Belmont Science Undergraduate Research Symposium (SURS):


“Blocking of thermotaxis and chemotaxis in Caenorhabditis elegans
laurynLauryn Bouldin
Faculty advisor: Robert Grammer, Ph.D.

Caenorhabditis elegans experience thermotaxis and chemotaxis. When the temperature is within the range of success for this organism but there is also a chemical repellent in the environment, it has to choose which stimulus is most important for it to respond to. The same is required when presented with an unfavorable temperature and a chemical attractant. Knowing what stimulus has the dominant response and that the organism is capable of overriding certain stimuli can be a reflection of how more complex organisms override temporarily noxious stimuli. This experiment tests wild-type C. elegans with benzaldehyde as a repellent and almond extract as the attractant while on a temperature gradient of favorable and unfavorable temperatures. It appears C. elegans choose to chemo tax toward almond extract and away from benzaldehyde even if it means thermo taxing toward an aversive temperature.

Phenotypic variation, movement and abundance of Vaejoviscarolinianusin a middle Tennessee cedar glade”
dillonC. Dillchelseaon Oman and Chelsea J. Lee
Faculty Advisor:  C. Steven Murphree, Ph.D.

Rocks harboring Vaejovis carolinianus in a middle Tennessee cedar glade were marked with flags.  The gender, weight, length measurements, number of pectinal teeth and carapace color were recorded. Each scorpion was marked on its dorsum with fingernail polish before being returned to its habitat.  During subsequent visits, marked rocks, along with rocks within a 10 m radius, were inspected for the presence of scorpions. Data were recorded for all recaptured individuals to estimate population size and these results, along with the range of measurements recorded for V. carolinianus, will be presented.

“The effects of anxiolytic medication on memory formation in stress induced Danio rerio specimens”

iqraIqra Wahid
Faculty Advisor: Dr. Lori McGrew, Ph.D.

Research with the study of zebrafish have led to advancements in developmental biology, oncology, toxicology, studies dealing with reproductive capabilities, environmental science, neurobiology, stem cell research, genetics and many other areas as well. Zebrafish (Danio rerio) have become one of the top research organisms because of the great list of model characteristics that the species possess’ and versatility in labs. Zebrafish, in the field of neuroscience, are being used to study how the brain encodes vision, hearing, movement, and even anxiety. By monitoring the neurons, activity within the zebrafish can be tracked and mapped out. In this experiment, zebrafish will be used to see the effects of the anxiolytic drug, Bupropion (commercially known as Wellbutrin), on spatial memory formation. They will be exposed to different doses of bupropion, in a pilot study in order to gauge a safe dosage level that could be used to treat the zebrafish. After administering a safe dosage to the zebrafish, the behavioral effects will be observed through the Noldus software that will track spatial movement of the zebrafish in the T-maze

“Effects of Win 55 212-2 Mesylate on Zebrafish Memory and Anxiety”
cassieCassie Michelle Wyatt
Faculty Advisor: Lori McGrew, Ph.D.

Cognitive functioning has been found to represent numerous brain mechanisms such as, memory association, pattern recognition, and problem solving. This functioning is highly active during the critical period of a life span. Once the critical period is over, however, cognitive decline can occur resulting in a variety of potential disorders, such as anxiety and Alzheimer’s. Zebrafish are a model organism used effectively in drug testing to determine relationships between the induced drug and the effects on cognitive functioning. WIN 55, 212-2 Mesylate is a nanomolar affinity cannabinoid receptor agonist. This study examines the effects of WIN 55, 212-2 mesylate on the memory, as well as the anxiety, of groups of dosed zebrafish using a T-Maze. It is expected that the dosed groups of zebrafish will have a slight decrease in their cognitive functioning; however, they will also have a decrease in anxiety. If so, this drug may be a potential replacement for other, harmful and sometimes deadly, prescription anxiety medications.