Title Information
Title
A New Model of Amyotrophic Lateral Sclerosis in Drosophila melanogaster: Precise Genetic Engineering, Characterization and Genetic Suppression
Name: Personal
Name Part
Sahin, Asli
Role
Role Term: Text
creator
Origin Information
Copyright Date
2015
Physical Description
Extent
xvii, 342 p.
digitalOrigin
born digital
Note
Thesis (Ph.D. -- Brown University (2015)
Name: Personal
Name Part
Reenan, Robert
Role
Role Term: Text
Director
Name: Personal
Name Part
Fallon, Justin
Role
Role Term: Text
Reader
Name: Personal
Name Part
Hart, Anne
Role
Role Term: Text
Reader
Name: Personal
Name Part
Wharton, Kristi
Role
Role Term: Text
Reader
Name: Personal
Name Part
Bonini, Nancy
Role
Role Term: Text
Reader
Name: Corporate
Name Part
Brown University. BIOMED: Molecular Biology, Cell Biology, and Biochemistry
Role
Role Term: Text
sponsor
Type of Resource
text
Genre (aat)
theses
Abstract
ABSTRACT Amyotrophic Lateral Sclerosis (ALS), the most common adult-onset motor neuron disease in the world, can be caused by mutations scattered along the full length of the Copper-Zinc Superoxide Dismutase 1 (SOD1) gene. Although ALS was characterized two decades ago as progressive muscle weakness due to extensive loss of motor neurons, the mechanisms underlying this incurable disease still remain elusive. More than 150 different point mutations have been found in the 153 amino acid encoding SOD1 gene of ALS patients. Despite substantial investigations in current SOD1-ALS animal models and post-mortem tissue analyses, the cellular mechanisms by which mutant SOD1 protein acquires toxic function are not well understood. One of the strongest hypotheses in the field suggests that aggregation due to misfolded SOD1 mediates cellular cytotoxicity. On the other hand, oxidative stress seems to play a critical role in ALS pathogenesis, since some transgenic models, especially those with low transgene copy number, do not develop disease symptoms unless they are challenged by reactive oxygen species. To address concerns about the dosage of mutant SOD1 in disease pathogenesis, we have genetically engineered four human ALS-causing SOD1 point mutations (G37R, H48R, H71Y and G85R) into the endogenous locus of Drosophila SOD1 (dSod1) via ends-out homologous recombination (HR); and analyzed the molecular, biochemical and behavioral alterations that arise from these mutations. Contrary to previous transgenic models, we have recapitulated ALS-like phenotypes in the fast ALS progression-causing SOD1 mutants without overexpressing the mutant protein. Drosophila carrying mutations causing fast disease progression in ALS patients (G85R and H71Y) exhibited neurodegeneration, locomotion deficit, and shortened life span whereas Drosophila carrying mutations causing slow disease progression in ALS patients (G37R and H48R) were indistinguishable when compared to controls. Next, we expanded characterization of ALS-like phenotype in Drosophila by performing a detailed expression profile from dSod1 mutants using deep sequencing technologies. Lastly, we performed a large-scale genetic screen aiming to identify suppressors to reverse lethality resulting from severe ALS mutations in Drosophila. This thesis work demonstrates a major step forward in our understanding of the molecular mechanisms and suppressors of mutant SOD1 when a precisely controlled system is studied.
Subject
Topic
ALS
Subject
Topic
Motor neuron
Subject
Topic
Disease
Subject
Topic
neurodegeneration
Subject
Topic
homologous recombination
Subject
Topic
ems
Subject
Topic
genetic screen
Subject (FAST) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/898388")
Topic
Drosophila
Subject (FAST) (authorityURI="http://id.worldcat.org/fast", valueURI="http://id.worldcat.org/fast/940027")
Topic
Genetic engineering
Record Information
Record Content Source (marcorg)
RPB
Record Creation Date (encoding="iso8601")
20160629
Language
Language Term: Code (ISO639-2B)
eng
Language Term: Text
English
Identifier: DOI
10.7301/Z0MG7MXP