Identifying Genes Regulating Synaptic
Development
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Masha Gelfand |
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The formation of synapses in complex organisms is an interesting yet poorly understood process.
The focus of the DiAntonio laboratory is to determine what impact postsynaptic cells have on presynaptic development. The model that is used is the neuromuscular junction of Drosophila (fruit fly). In previous research, the laboratory has identified a group of genes that when overexpressed in all muscles, is lethal to the fruit fly. The goal of my research has been to cross two lines of flies in order to overexpress each of these genes in only two particular muscles in the fly larvae. If the genes are overexpressed in only two muscles, the genes are not lethal, but their effects are still clearly seen in the two muscles.
Once I had crossed the lines, I had to look at the synapses to see whether they were normal. In order to accomplish this I stained the resulting flies using two different immunostaining procedures. The first used a rabbit alpha-HRP stain, designed to give a picture of the synapse. The second stain was a rabbit alpha -GluRC stain, which showed the neurotransmitter receptor structure. The purpose of the alpha -HRP stain was to observe the general appearance of the synapse and boutons. Several questions I asked were: Are the synapses more branched than usual? Are there more boutons than normal? Are the boutons a reasonable size, and are they distributed correctly? The purpose of the alpha -GluRC stain was to observe the neurotransmitter receptor distribution as compared to wild-type specimens.
I crossed and analyzed 20 lines of flies. The results that I observed were expected. Out of the 20 lines, one may have looked mutant. The alpha -HRP stain revealed a very branched structure with many underdeveloped boutons, while the alpha -GluRC stain showed normal neurotransmitter receptor distribution. This project would require more experiments to determine whether the irregularity observed was due to the overexpressed gene.
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Washington University - Biology
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