Date of Award

1-1-2011

Document Type

Masters Thesis

Degree Name

M.S.

Organizational Unit

Biological Sciences

First Advisor

Scott A. Barbee, Ph.D.

Second Advisor

Joseph Angleson

Third Advisor

Robert M. Dores

Keywords

Activity dependent, Ghost bouton, MicroRNA, miRNA, Neuromuscular junction, Quantitative polymerase chain reaction

Abstract

The primary goal of this thesis was to determine if spaced synaptic stimulation induced the differential expression of microRNAs (miRNAs) in the Drosophila melanogaster central nervous system (CNS). Prior to attaining this goal, we needed to identify and validate a spaced stimulation paradigm that could induce the formation of new synaptic growth at a model synapse, the larval neuromuscular junction (NMJ). Both Channelrhodopsin- and high potassium-based stimulation paradigms adapted from (Ataman, et al. 2008) were tested. Once validation of these paradigms was complete, we sought to characterize the miRNA expression profile of the larval CNS by miRNA array. Following attainment of these data, we used quantitative real-time PCR (RT-qPCR) to determine if acute synaptic stimulation caused the differential expression of neuronal miRNAs. We found that upon high potassium spaced training in a wild type (Canton S) genotype, 5 miRNAs showed significant differential expression when normalized to a validated reference gene, the U1 snRNA. Moreover, absolute quantification of our RT-qPCR study implicated one miRNA: miR-958 as being significantly regulated by activity. Investigation into potential targets for miR-958 revealed it to be a potential regular of Dlar, a protein tyrosine phosphatase implicated in synapse development. This investigation provides the foundation to directly test our underlying hypothesis that, following spaced training, differential expression of miRNAs alters the translation of proteins required to induce and maintain these structural changes at the synapse.

Publication Statement

Copyright is held by the author. User is responsible for all copyright compliance.

Rights Holder

Robert Ian Sand

Provenance

Received from ProQuest

File Format

application/pdf

Language

en

File Size

93 p.

Discipline

Neurosciences, Biology, Molecular Biology

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