Honors Theses

Date of Award

2007

Document Type

Undergraduate Thesis

Department

Chemistry and Biochemistry

First Advisor

Bradley Jones

Relational Format

Dissertation/Thesis

Abstract

Complex nervous systems require both neurons and glial cells for proper functioning. While neurons have been the primary focus of neuroscience research, much more attention is now being demanded by the glia, whose numerous unknown functions become clearer. Studies on Drosophila melanogaster have been key to our understanding of the nervous system, as the development of both neurons and glia has been well characterized in this organism. In Drosophila, the effects of manipulating the nervous system can be readily observed using immunostaining of different cell types. This study examined the consequences of expressing reaper {rpr) in the most medial cell body glia (MM-CBG) and the medial cell body glia (M-CBG), a subset of glial cells. Because rpr is a gene that initiates programmed cell death (PCD), the intent was to develop a tool to express rpr in these specific glial cells. By using PCD to eliminate the MM-CBG and M-CBG and then observing the effects of this cell death on subsequent nervous system development, the fimctions of these cells can be deduced. Crosses were carried out between glial-specific GAL4 lines and a UAS-/flcZ line to demonstrate that gene expression is targeted to the MM-CBG and M-CBG. Once verified, a line was manipulated so that it contained both the GAL4 element and UASlacZ. This new line was then crossed to two UAS-/y?r lines, UAS-r/^r*"^ and UAS-rpr^^ to see if the Reaper protein killed the cells. Crosses with wild-type flies were also done as a control. For all of the crosses, embryos were collected on apple juice plates and fixed with formaldehyde before utilizing antibody staining to observe where the gene is expressed. Ectopic expression of rpr^^ in the MM-CBG and M-CBG did cause programmed cell death in the cells, whereas ectopic expression of rpr^^ did not kill the cells. The results indicate that ectopic rpr expression is sufficient to kill the MM-CBG and M-CBG. The fact that only one UAS-rpr line worked is most likely due to differences in the amount of expression of the Reaper protein. The local chromosomal environment in which each of the constructs is located could cause this kind of dosage difference.

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