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MOLECULAR MECHANISMS UNDERLYING CADHERIN-6B INTERNALIZATION IN PREMIGRATORY CRANIAL NEURAL CREST CELLS DURING THEIR EPITHELIAL-TO-MESENCHYMAL TRANSITION

dc.contributor.advisorTaneyhill, Lisa Aen_US
dc.contributor.authorPadmanabhan, Rangarajanen_US
dc.date.accessioned2015-06-25T05:43:10Z
dc.date.available2015-06-25T05:43:10Z
dc.date.issued2015en_US
dc.identifierhttps://doi.org/10.13016/M22G9H
dc.identifier.urihttp://hdl.handle.net/1903/16464
dc.description.abstractThe generation of migratory cells from immotile precursors occurs frequently throughout development and is crucial to the formation and maintenance of a functioning organism. This phenomenon, called an epithelial-to-mesenchymal transition (EMT), involves the disassembly of intercellular adhesions and cytoskeletal rearrangements in order to promote migration. Importantly, aberrant EMTs and cell migration can lead to devastating human conditions including cancer metastasis and fibrosis. How cells accomplish EMT to become migratory is still an unanswered question in the biomedical field. To this end, we use chick neural crest cells as an in vivo model to elucidate the molecules and pathways that regulate EMT and migration. Neural crest cells are a population of embryonic cells that are originally stationary within the dorsal neural tube but later migrate to form a variety of adult derivatives, such as the craniofacial skeleton, skin pigment cells and portions of the heart. To facilitate EMT, chick premigratory neural crest cells lose intercellular contacts mediated, in part, by the transmembrane cell adhesion protein Cadherin-6B (Cad6B). While Cad6B mRNA is transcriptionally repressed in premigratory neural crest cells, loss of Cad6B protein does not directly follow and instead occurs ~90 minutes later, just prior to migration. This rapid depletion of Cad6B is all the more striking given that the half-life of most cadherins, including Cad6B, is ~6-8 hours in vitro. As such, unique post- translational mechanisms must exist to remove Cad6B from premigratory neural crest cell plasma membranes to facilitate neural crest EMT. Since cadherins are known to be downregulated through internalization mechanisms (e.g., endocytosis, macropinocytosis) in other in vitro systems, the hypothesis of this dissertation is that Cad6B is internalized, and that this process plays a critical function to enable neural crest EMT. To this end, we document the existence of Cad6B cytoplasmic puncta in cultured cells, cultured neural crest cells and transverse sections of chick embryos. We subsequently identified a p120-catenin binding motif in the Cad6B cytoplasmic tail and demonstrated its functionality through site-directed mutagenesis, revealing a role in enhancing Cad6B internalization and reducing the stability of membrane-bound Cad6B. Furthermore, we uncover for the first time that Cad6B is removed from premigratory cranial neural crest cells through cell surface internalization events that include clathrin-mediated endocytosis and macropinocytosis. Both of these processes are dependent upon the function of dynamin, and inhibition of Cad6B internalization abrogates neural crest cell EMT and migration. Collectively, our findings provide a molecular blueprint for how cadherins are dynamically regulated during the formation of migratory cell types required for normal embryonic development and tissue repair as well as those generated during human diseases and cancers. Importantly, our research is multi-disciplinary, integrating cell biology and physiology to reveal how a cellular event, the active downregulation of a membrane protein, results in a physiological event, neural crest EMT and migration.en_US
dc.language.isoenen_US
dc.titleMOLECULAR MECHANISMS UNDERLYING CADHERIN-6B INTERNALIZATION IN PREMIGRATORY CRANIAL NEURAL CREST CELLS DURING THEIR EPITHELIAL-TO-MESENCHYMAL TRANSITIONen_US
dc.typeDissertationen_US
dc.contributor.publisherDigital Repository at the University of Marylanden_US
dc.contributor.publisherUniversity of Maryland (College Park, Md.)en_US
dc.contributor.departmentAnimal Sciencesen_US
dc.subject.pqcontrolledCellular biologyen_US
dc.subject.pqcontrolledDevelopmental biologyen_US
dc.subject.pqcontrolledMolecular biologyen_US
dc.subject.pquncontrolledCadherinen_US
dc.subject.pquncontrolledCanceren_US
dc.subject.pquncontrolledEmbryoen_US
dc.subject.pquncontrolledEMTen_US
dc.subject.pquncontrolledEndocytosisen_US
dc.subject.pquncontrolledNeural cresten_US


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