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
Daniel Linseman
Third Advisor
Todd Blankenship
Fourth Advisor
Martin Margittai
Keywords
Microtuble, Futsch, Neuron, Micro-RNA
Abstract
Fragile X syndrome (FXS) is the most common form of inherited mental retardation in humans. FXS is caused by loss of the Fragile X Mental Retardation Protein (FMRP), an important regulator of neuronal mRNA translation. Patients with FXS display cognitive deficits including memory problems. Protein synthesis-dependent long-term changes in synaptic plasticity are involved in the establishment and maintenance of long-term memory. One prevalent theory of FXS pathology predicts that FMRP is required to negatively regulate the translation of important mRNAs at the synapse. We are investigating microRNAs (miRNAs) as a potential regulator of synaptic FMRP-regulated mRNAs that have previously been described as being crucial to the process of synaptic plasticity.
The general hypothesis underlying this thesis is that FMRP may negatively regulate the expression of futsch (the Drosophila homologue of the microtubule-associated protein gene MAP1B) via the miRNA pathway. The first step we took in testing this hypothesis was to confirm that futsch is subject to miRNA-mediated translational control. Using in silico target analysis, we predicted that several neuronally expressed miRNAs target the futsch mRNA 3'UTR and repress expression of Futsch protein. Then, using an in vitro luciferase reporter system, we showed that miR-315 and members of the miR-9 family selectively down-regulated futsch reporter translation. We have confirmed by site-directed mutagenesis that the miRNA interaction with the futsch 3'UTR is specific to the miRNA seed region binding site. Interestingly, reduction of FMRP levels by RNAi had no effect on futsch 3'UTR reporter expression. Together, these data suggest regulation of futsch expression by the miRNA pathway might be independent of FMRP activity. However, additional experiments need to be completed to confirm these preliminary results.
Publication Statement
Copyright is held by the author. User is responsible for all copyright compliance.
Rights Holder
Leslie M. Rozeboom
Provenance
Received from ProQuest
File Format
application/pdf
Language
en
File Size
66 p.
Recommended Citation
Rozeboom, Leslie M., "MicroRNAs 9a, 9b, 9c and 315 Regulate Expression of a Reporter for the Neuronal Microtubule-Associated Protein Futsch/MAP1B" (2011). Electronic Theses and Dissertations. 914.
https://digitalcommons.du.edu/etd/914
Copyright date
2011
Discipline
Biology, Neurosciences