Investigating the Role of Olfm1 in the Trafficking of GluR2-containing AMPA receptors
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Authors
Cardenas, Caroline
Date of Issue
2015-04-01
Type
thesis
Language
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Abstract
Neurons have the ability to either increase or decrease the strength of their connections with other neurons. This property is known as synaptic plasticity and is thought to influence higher cognitive functions such as learning and memory. Decreases in cognitive abilities as well as neurological disorders can be attributed to decreased synaptic plasticity. The variable expression of GluR2-containing AMPARs has been linked to changes in synaptic plasticity. The mechanism governing the trafficking of GluR2-containing AMPARs to synapses is not fully understood and was the main focus of my study. I hypothesized that Olfm1 (Olfactomedin 1) plays a key role in the trafficking of GluR2-containing AMPARs to the synapse. HA-tagged Olfm1 was successfully cloned into the mammalian expression vector pcDNA3.1. Western blot analysis verified Olfm1 protein expression in both transfected HEK293 cells as well as endogenous expression in NT2 neurons. GluR2-GFP protein expression was seen in transfected HEK293 cells, but we were unable to confirm endogenous expression in the NT2 neuron cultures. Co-immunoprecipiation experiments were also performed to study the interaction of Olfm1 and GluR2, but the tests remain inconclusive. Therefore, further testing is needed in order to accept or reject my hypothesis that Olfm1 plays a role in the trafficking of GluR2-containing AMPARs.