Characterization of the Molecular Mechanisms by which Endocytosed AMP AR Mediates NMDAR- Mediated Synaptic Plasticity and Excitotoxicity
Antonuk, C. Danielle
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Strokes are the leading cause of serious, long-term disabilities in the United States. When a stroke occurs the brain is deprived of oxygen and glucose, which triggers various pathways that cause the apoptosis and necrosis of neurons. We were interested in examining one pathway that leads to apoptosis involving the interactions of the GLUR2 subunit of the a-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor (AMPAR) after it is internalized by N-methyl-D-aspartate (NMDA) excitotoxicity. Using Rattus norvegicus cortex neuron cells that had undergone NMDA stimulation or oxygen-glucose deprecation (OGD), both of which trigger the NMDA- AMP AR relationship, GLUR2 was examined. Significant headway was made in determining the result of the endocytosis of GL UR2 from AMP AR on the post-synaptic cell membrane. This included identifying an SP 1 on GluR2 promoter region binding site, the development of several probes for future gel shift assays, and the synthesis of NR2A and NR2B proteins which will be used to make antibodies. Further study will be required to answer the many remaining questions such as the specific binding sequence of protein GLUR2 and SPl, and the effect of NMDA excitotoxicity on GluR2 mRNA and protein levels. This project, however, served to develop the tools necessary to carry out these next steps.
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