Presentation Title

Using Drosophila Reporters to Evaluate Metal Uptake, ROS Production, and Chelation in Drosophila S2 Cells and Human HeLa and Neuroblastoma Cells

Start Date

November 2016

End Date

November 2016

Location

HUB 302-#33

Type of Presentation

Poster

Abstract

The goal of our project is to determine the rates of metal uptake and reactive oxygen species (ROS) production in Drosophila S2 cells and human HeLa and neuroblastoma cells using a Drosophila metal- and ROS-sensitive luciferase reporter. An additional goal was to use these reporters to examine the effects of metal chelation in ROS production in these cells since metal chelators are suggested as potential therapeutics for neurodegenerative diseases. First, we performed time course experiments to estimate the rates of metal uptake and ROS production in S2 cells by measuring the luciferase signals from the Mtn and SOD promoters after treating the cells with Cu, Fe, and Zn for different time periods. From this study, we found that the luciferase signal from the Mtn promoter construct increased linearly over time when the cells were treated with Cu and Zn, but not with Fe. This trend was also observed with the SOD promoter construct. For our chelation tests, we used EDTA, citrate, and histidine because they have been suggested as potential therapeutics for neurodegenerative diseases. We performed the chelation experiments in Drosophila S2 cells and human HeLa and SHSY cells by measuring the luciferase activities from the Mtn and SOD promoters in cells treated either with Cu, Fe, or Zn in the presence or absence of EDTA, histidine, or citrate. Of the nine metal-chelator combinations tested in S2 cells, the only combination that led to a significant decrease in Mtn-luciferase activity was Zn and citrate , which correlated with a corresponding decrease in SOD-luciferase activity. The same effect was observed in HeLa and SHSY-5Y cells with the Mtn promoter construct. Additionally, we found that in HeLa cells treated with Cu and histidine, there was a 2-fold decrease in Mtn-luciferase activity relative to cells treated with histidine alone.

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Using Drosophila Reporters to Evaluate Metal Uptake, ROS Production, and Chelation in Drosophila S2 Cells and Human HeLa and Neuroblastoma Cells

HUB 302-#33

The goal of our project is to determine the rates of metal uptake and reactive oxygen species (ROS) production in Drosophila S2 cells and human HeLa and neuroblastoma cells using a Drosophila metal- and ROS-sensitive luciferase reporter. An additional goal was to use these reporters to examine the effects of metal chelation in ROS production in these cells since metal chelators are suggested as potential therapeutics for neurodegenerative diseases. First, we performed time course experiments to estimate the rates of metal uptake and ROS production in S2 cells by measuring the luciferase signals from the Mtn and SOD promoters after treating the cells with Cu, Fe, and Zn for different time periods. From this study, we found that the luciferase signal from the Mtn promoter construct increased linearly over time when the cells were treated with Cu and Zn, but not with Fe. This trend was also observed with the SOD promoter construct. For our chelation tests, we used EDTA, citrate, and histidine because they have been suggested as potential therapeutics for neurodegenerative diseases. We performed the chelation experiments in Drosophila S2 cells and human HeLa and SHSY cells by measuring the luciferase activities from the Mtn and SOD promoters in cells treated either with Cu, Fe, or Zn in the presence or absence of EDTA, histidine, or citrate. Of the nine metal-chelator combinations tested in S2 cells, the only combination that led to a significant decrease in Mtn-luciferase activity was Zn and citrate , which correlated with a corresponding decrease in SOD-luciferase activity. The same effect was observed in HeLa and SHSY-5Y cells with the Mtn promoter construct. Additionally, we found that in HeLa cells treated with Cu and histidine, there was a 2-fold decrease in Mtn-luciferase activity relative to cells treated with histidine alone.