Margaret Werd
Pronouns: she/her
Research Mentor(s): Nan Hatch
Co-Presenter:
Research Mentor School/College/Department: Orthodontics and Pediatric Dentistry / Dentistry
Presentation Date: April 20
Presentation Type: Poster
Session: Session 4 – 2:40pm – 3:30 pm
Room: League Ballroom
Authors: Emmy Werd, Erica Siismets, Nan Hatch
Presenter: 60
Abstract
Humans and mice with Crouzon craniosynostosis syndrome have distinct craniofacial features including dome-shaped skulls, wide-set bulging eyes, premature fusion of coronal and facial sutures, and a severely retrusive midface. Surgical intervention is the only available treatment. Crouzon syndrome is caused by mutations in the FGFR2 gene. It remains unknown how these mutations cause the distinct craniofacial phenotype. The goal of this study is to investigate how Crouzon FGFR2C342Y protein is processed and trafficked inside cells. We have previously shown using Western blot technique that the Crouzon mutant FGFR2 exhibits immature/incomplete glycosylation. We hypothesize that the mutant receptor is degraded more than the WT receptor due to this immature/incomplete glycosylation. In order to visualize whether incomplete protein glycosylation affects the intracellular trafficking and/or degradation of FGFR2C342Y, MC3T3 (E1) mouse cranial bone cells were transiently transfected with wild type (WT) or mutant FGFR2 plasmids that contained a special sequence to anchor the protein to the endoplasmic reticulum. Biotin was added to the cell culture, which then caused the ER to release the plasmid into the protein secretory pathway. Fluorescent images of the cells were obtained using a Zeiss Axio Microscope. Results show decreased fluorescent expression of Crouzon mutant FGFR2 compared to WT after 30 minutes of biotin exposure. This suggests that the Crouzon mutant FGFR2 exhibits earlier and potentially more degradation compared to WT FGFR2. Next steps will include using immunofluorescence technique to visualize localization of WT and mutant FGFR2 in various organelles (Golgi apparatus, lysosomes, etc.) as well as using Western blot technique to investigate FGFR downstream signaling. This future work will further advance our understanding of the intracellular effects of Crouzon mutant FGFR2 and how this possibly leads to the Crouzon syndrome craniofacial phenotype.
Biomedical Sciences, Interdisciplinary, Natural/Life Sciences
