Throughout my summer in the PURM program, I worked in Dr. Adeline Vanderver’s lab in the Leonard and Madlyn Abramson Pediatric Research Center. Within her lab, I assisted her project on in vivo modelling of H-ABC and worked closely with Dr. Sunetra Sase and Dr. Akshata Almad. The project’s goal is to create a model of the severe TUBB4A-related leukodystrophy H-ABC using mice, since there is no current model of the disease.
H-ABC stands for hypomyelination and atrophy of the basal ganglia and cerebellum. It results from a mutation of the TUBB4A gene, which encodes instructions for creating beta-tubulin proteins. The beta-tubulin protein is very prominent in the basal ganglia, cerebellum, and white matter of the brain. Symptoms of the disease usually begin to show during early childhood. Symptoms include motor dysfunction, delayed mental development, ataxia, and seizures. With H-ABC’s recent discovery, the number of known individuals affected by this disease continues to increase. Creating a model provides better understanding of the disease and creates an essential matrix for future experiments testing treatments.
Throughout the summer, I have learned numerous lab skills. I have learned how to work with mice and various behavioral tests, including righting-reflex, Rota-rod, ambulatory angle, and ambulation tests. I also learned how to tattoo, anesthetize, perfuse, and genotype the mice. Working with mice presented a challenge the first few weeks, as I had no prior experience with lab animals. After daily acclimation, I learned how to handle them with ease and how to have them perform tests. I also learned how to analyze the data recorded from these tests. For example, after recording ambulation videos of each mouse, I would watch the videos to score their movement accurately according to a chart. I also learned how to measure hind limb foot angles of the mice according to the created criteria. The data from these tests were then used to compare the behavior of mice from various genotypes and ages in order to assess the effect of the TUBB4A mutation on behavior.
Furthermore, I learned how to perform anti-neuronal nuclei (NeuN) antibody immunostaining on brain sections of mice of various ages and genotypes. I used a cell counter to perform NeuN count on the images taken of the striata. I learned how to cut coronal and sagittal brain sections using the Cryostat, which I realized was a skill that required a lot of patience and meticulous attention to detail.
I had an amazing summer experience through the PURM program and have learned so much. Not only did I learn various important lab skills, but I was also able to immerse myself into the field of neuroscience. I plan on continuing in Dr. Vanderver’s lab on her H-ABC project throughout the school year, and I am excited for the future work planned!