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gilinternship

Characterizing the CB1 Cannabinoid Receptor Using TRUPATH Assays at RTI International – with Nivetha Ramasamy

Updated: Aug 24


Hi! My name is Nivi Ramasamy and I’m a junior from Charlotte, NC. I’m currently pursuing a B.S. in Neuroscience and a B.S.P.H. in Nutrition – Science and Research track with a minor in Chemistry. I have always been fascinated by the brain, with my interests lying primarily in cognitive and developmental neuroscience along with neural networks. Over the last year, I’ve also gained a newfound interest in the role that nutrition plays in public health conditions.


My first research experience was in high school at East Carolina University through the NCSSM Summer Ventures program. After intubating rat pups with ethanol/milk solutions to induce Fetal Alcohol Syndrome (FAS), I analyzed the adult rat brains after euthanasia to determine the effects of FAS on cell viability in the interpositus nucleus of the cerebellum. This experience introduced me to the exciting world of neuroscience research and I knew when I arrived at UNC that I wanted to get involved in more research as soon as I could. During the fall of my freshman year, I joined the Frohlich lab, which is a psychiatry lab in UNC’s School of Medicine, and I have been a research assistant there for the last two years. I study sustained attention and electrophysiological processes in the ferret model by running a 5-choice serial reaction time task and transcranial alternating current stimulation sessions on the ferrets, performing neurohistology, and conducting behavioral data analysis using MATLAB. In order to explore my other research interests outside neuroscience, I also joined the Hursting lab at the UNC Gillings School of Global Public Health earlier this year. In this lab, I explore the connection between obesity-associated inflammation in adipose tissue and colorectal cancer using mice and cell culture.


While I found neuropharmacology interesting in course settings, it was not an area of neuroscience that I saw myself researching until the Gil internship. When I heard that one of my worksite options was a neuropharmacology lab focused on molecular cannabis research, I was excited to explore a new area of research, but also nervous about my lack of pharmacology research experience. However, I had great mentors at my internship worksite and transferable skills from my prior research experiences that helped me adjust to the new research environment I was placed in. My internship worksite this semester was at RTI International, a non-profit research institute located in Research Triangle Park, NC. I had the opportunity to work with Dr. Thomas Gamage (principal investigator) and Daniel Barrus (chemist and my direct mentor in the lab) in the Analytical Chemistry and Pharmaceuticals department. This semester, I studied how different synthetic cannabinoids, phytocannabinoids, endocannabinoids, and allosteric modulators affected the CB1 cannabinoid receptor by running a type of bioluminescence resonance energy transfer assays (BRET) called the TRUPATH assay. The TRUPATH assay consists of biosensors that allow us to characterize G-protein-coupled receptors (GPCRs) by directly changing the α, β and γ subunits of the GPCR.


In order to run the experiments, I first passage and seed dishes with human embryonic kidney 293 (HEK-293) cells. The next day, I transfect the cells with the hCB1 or hNPBWR1 receptor and our GPCR subunits of interest. 24 hours after that, I plate the transfected cells in 96-well plates that will be used when running the assay. On the fourth day, I treat the cells with our drugs of interest and then run the assay. Besides gaining hands-on experience with cell culture and molecular neuropharmacology research, I have also learned a lot about pharmacology and data analysis through my mentors. Daniel, who is my direct mentor in the lab and also a previous Gil intern, regularly goes through our data and the graphs of our results with me. He has helped me understand how BRET ratios from the experiments we run demonstrate whether our drugs are agonists or antagonists and how different drugs affect our receptors of interest over time for kinetic experiments. Dr. Gamage also meets with me to talk about pharmacology concepts (like logarithmic dose-response relationships) and discuss how different forms of data analysis (such as curve fitting) allow us to look at our data in the most comprehensive way.


I’m very thankful to both Dr. Gamage and Daniel for all their support this semester and their time spent training and teaching me. I’m also thankful to both Dr. Buzinski and Chelsea for giving me this opportunity and supporting me in the development of my professional skills through our weekly classes and activities. The Gil Internship has been an invaluable experience for me by allowing me to take part in research outside of a university-setting and exposing me to a new kind of research that I can see myself potentially researching in the future. After finishing my undergraduate studies, I hope to attend graduate school and pursue a Ph.D., and I believe that the Gil internship has been an important stepping stone in my journey to get there.


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