Effects of Glutamate Receptor Activation on Gld2-Dependent Synthesis of CaMKIIα and NMDA Receptor Subunits Open Access

He, Yuncen Atticus (2012)

Permanent URL: https://etd.library.emory.edu/concern/etds/f1881m27r?locale=en
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Abstract

The alteration of protein compositions at synapses following receptor stimulation is a crucial event that is thought to underlie processes such as memory storage and learning. However, the mechanisms behind this regulation and the functional units involved are not well known. Studies pointing to a group of proteins encoded by mRNAs possessing cis-acting elements that interact with mRNA binding proteins like the cytoplasmic element binding protein (CPEB) provide us with motivation to investigate the regulation of postsynaptic protein subunits such as the α-subunit of Ca2+/calmodulin-dependent protein kinase (CaMKIIα) and the 2A subunit of N-methyl-D-aspartate receptors (NR2A) after activation of glutamate receptors. Based upon preliminary data from the Bassell Lab, we hypothesize that the poly(A) polymerase Gld2 positively regulates activity induced synthesis of CaMKIIα and NR2A in dendrites and that chemical long-term potentiation (LTP) stimulation will induce protein synthesis-dependent insertion of NR2A in the plasma membrane. To assess our propositions, we stimulated hippocampal neurons with either glutamate or a chemical-LTP paradigm and observed the effects of the stimulations on CaMKIIα and NMDAR subunits through immunocytochemistry. Anisomycin, a protein synthesis inhibitor, and Gld2 short hairpin RNA (shRNA) lentiviruses were used to investigate the Gld2-dependent protein synthesis of these subunits. From this study, we determined that there is strong evidence suggesting glutamate stimulation increases the Gld-2 dependent expression of CaMKIIα and NR2A protein in distal dendritic regions. Although we did not observe significant changes in the surface NMDAR subunit expressions, we provide some suggestions for future experiments that could better support the second part of our hypothesis.

Table of Contents

Table of Contents
Introduction.......................................................................................................................1
Methods............................................................................................................................7
Results............................................................................................................................12
Discussion........................................................................................................................19
References.......................................................................................................................29
Figures............................................................................................................................32

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