Identifying genes required for axonal maintenance in Drosophila melanogaster using an EMS forward MARCM genetic screen
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2022-10-12
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en
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Abstract
As the life expectancy of the world increases, so does the prevalence of neurodegenerative diseases.
Parkinson’s disease and Alzheimer’s disease are the most common. Motor neurodegenerative
diseases such as amyotrophic lateral sclerosis are also branches of these diseases, but are rare. The
cause of most neurodegenerative diseases remains unknown, but identifying the genes involved
would provide us with knowledge about their mechanistic origins. In this study, we performed an EMSbased
forward MARCM genetic screen of the X-chromosome of Drosophila melanogaster. This
technique is hypothesis- and prediction-free, meaning there is no a priori knowledge about what
genes or mechanisms might be involved. The EMS treatment given to male Drosophila resulted in
8742 viable fly stocks, all with a mutation on their X-chromosome. Screening and analysis revealed
1452 hemizygously lethal stocks and 131 stocks with a range of neurodegenerative phenotypes. Out
of these 131 stocks, 58 stocks had cell lethal phenotype and 73 stocks had a visible neurodegenerative
phenotype. From these 73 stocks, duplication mapping revealed 46 stocks rescued by a genomic
duplication region and 27 stocks remaining to be rescued. Complementation mapping revealed 8
complementation groups with mutations in the same gene. 9 out of the 46 rescued stocks were also
rescued by a P[acman] duplication line which identifies a small region on the X-chromosome where
the mutation is located. 7 mutant stocks underwent Illumina DNA Library Preparation and Illumina
Next Generation Sequencing. Once the remaining stocks have been analysed and chromosomal
mapping has been completed, this research can be taken forward with further DNA library preparation
whole genome sequencing, behavioural assays and human homologue comparison and be an ideal
stepping stone towards discovering new genes involved in neurodegenerative diseases
Keywords: neurodegeneration, axonal maintenance, Drosophila melanogaster, MARCM,
duplication mapping, complementation mapping, whole genome sequencing, EMS
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Faculteit der Sociale Wetenschappen