dc.creator | Marks, Christian Randal | |
dc.date.accessioned | 2020-08-21T21:02:46Z | |
dc.date.available | 2019-02-21 | |
dc.date.issued | 2019-02-21 | |
dc.identifier.uri | https://etd.library.vanderbilt.edu/etd-02202019-113322 | |
dc.identifier.uri | http://hdl.handle.net/1803/10598 | |
dc.description.abstract | Multi-protein complexes formed through protein-protein interactions in the dendrites of neurons are highly regulated to facilitate proper synaptic function. The work presented in this dissertation highlights the importance of a novel interaction between two synaptic regulators: Calcium/Calmodulin Dependent Protein Kinase II (CaMKII) and the metabotropic glutamate receptor 5 (mGlu5).
Here, I show that CaMKII can bind to and phosphorylate the mGlu5 C-terminal domain (CTD). In vitro characterization showed that the mGlu5-CTD directly interacts with Thr286-autophosphorylated CaMKII. I identified three basic residues on the mGlu5-CTD necessary for the mGlu5-CaMKII interaction. Mutagenesis of these residues allowed for the construction of a full-length mGlu5 construct with reduced CaMKII binding.
I hypothesized that the mGlu5-CaMKII interaction could modulate mGlu5 signaling. Activation of mGlu5 results in downstream signals that increase intracellular Ca2+ release and activate ERK to regulate many cellular processes. The co-expression of active CaMKII increased basal mGlu5 surface expression and ERK activation in heterologous cells. In addition, CaMKII modulated mGlu5-mediated Ca2+ release in heterologous cells, decreasing the initial Ca2+ amplitude, but prolonging the relative Ca2+ signal. Therefore, I hypothesized that knockdown of CaMKII in neurons would result in increased mGlu5-mediated Ca2+ signals. However, knockdown of CaMKII reduced mGlu5-specific Ca2+ signals in neuronal cultures. This effect was specific to mGlu5 because CaMKII knockdown had no effect on L-type voltage dependent Ca2+ channel (LTCC) signals. However, knockdown of an important synaptic scaffolding protein, Shank-3, reduced mGlu5 and LTCC Ca2+ responses. | |
dc.format.mimetype | application/pdf | |
dc.subject | kinase activity | |
dc.subject | calicum imaging | |
dc.subject | neuronal activity | |
dc.subject | camkii | |
dc.subject | protein interactions | |
dc.subject | metabotropic glutamate receptor | |
dc.subject | phosphorylation | |
dc.subject | protein binding | |
dc.title | Role of Ca2+/Calmodulin-Dependent Protein Kinase II in Regulating
the Metabotropic Glutamate Receptor 5 | |
dc.type | dissertation | |
dc.contributor.committeeMember | Eric Delpire, Ph.D. | |
dc.contributor.committeeMember | Colleen Niswender, Ph.D. | |
dc.contributor.committeeMember | David Jacobson, Ph.D. | |
dc.contributor.committeeMember | Brian Wadzinski, Ph.D. | |
dc.type.material | text | |
thesis.degree.name | PHD | |
thesis.degree.level | dissertation | |
thesis.degree.discipline | Molecular Physiology and Biophysics | |
thesis.degree.grantor | Vanderbilt University | |
local.embargo.terms | 2019-02-21 | |
local.embargo.lift | 2019-02-21 | |
dc.contributor.committeeChair | Danny Winder, Ph.D. | |