The main objectives of this proposal are to characterize the developmental profile of initiation and progression of CaMKII misregulation in a mouse model of Angelman Syndrome, establish the consequences of CaMKII misregulation on glutamate receptors, and to use viral overexpression approaches to prevent or rescue defects in synaptic transmission and/or behavior.
Specific Aim 1 involves determining the developmental time point when CaMKII misregulation occurs in AS mice. This will be done by determining the phosphorylation state and the activity levels of CaMKII through the use of western blot analysis and kinase activity assays to confirm what time point during development CaMKII phosporylation (T286 and T305) is increased and when activity of CaMKII is inhibited. The effects of CaMKII misregulation in AS on glutamate receptors will be determined by identifying changes in expression, subunit composition, and phosphorylation states of these receptors using western blot analysis. Additionally, changes in receptor function will be assayed using electrophysiological methods.
Specific Aim 2 will determine the therapeutic potential of genetic alteration of CaMKII activity. This will be done through injection of lentivirus expressing TT305/306AA CaMKII into hippocampal neurons in both in vitro and in vivo settings. Electrophysiological and behavioral techniques will be used to determine the therapeutic potential in rescuing the AS phenotype through this approach. This research proposal will determine the molecular mechanism underlying Angelman Syndrome (AS). This will lead to the identification of targets that can be manipulated through therapeutic intervention to treat and possibly cure AS. ? ? ?
| Gustin, Richard M; Shonesy, Brian C; Robinson, Stacey L et al. (2011) Loss of Thr286 phosphorylation disrupts synaptic CaMKII? targeting, NMDAR activity and behavior in pre-adolescent mice. Mol Cell Neurosci 47:286-92 |
| Gustin, Richard M; Bichell, Terry Jo; Bubser, Michael et al. (2010) Tissue-specific variation of Ube3a protein expression in rodents and in a mouse model of Angelman syndrome. Neurobiol Dis 39:283-91 |
| Baucum 2nd, Anthony J; Jalan-Sakrikar, Nidhi; Jiao, Yuxia et al. (2010) Identification and validation of novel spinophilin-associated proteins in rodent striatum using an enhanced ex vivo shotgun proteomics approach. Mol Cell Proteomics 9:1243-59 |
| Brigman, Jonathan L; Wright, Tara; Talani, Giuseppe et al. (2010) Loss of GluN2B-containing NMDA receptors in CA1 hippocampus and cortex impairs long-term depression, reduces dendritic spine density, and disrupts learning. J Neurosci 30:4590-600 |
