Title

Investigating Mitochondrial Dynamics and Cell Life Cycles through Induced Stress

Document Type

Poster

Campus where you would like to present

Ellensburg

Event Website

https://digitalcommons.cwu.edu/source

Start Date

18-5-2020

Abstract

Mitochondria are organelles that produce energy rich molecules called ATP through cellular respiration. Mitochondria have been implicated in numerous disease states and could be an attractive drug target. We hypothesize that mitochondrial shape, reactive oxygen species (ROS) generation, and cell death are interrelated. During respiration, ROS are produced and can react with cell components which causes damage. Stress induced by ROS likely causes mitochondrial shape change, and in extreme cases, leads to cell death. Cell death was investigated by using a WST-8 cell viability assay on cultured mouse liver cells exposed to the oxidative stressor, menadione, which produces ROS. It was found that increased concentrations of menadione led to increased cell death. To monitor mitochondrial shape changes, a protein-based fluorescent probe was used to mark mitochondria in cells. A mitochondrial green fluorescent protein (GFP) plasmid was transfected into the mouse liver cells, so that the cells could express the mitochondrial targeted fluorescent probe. Fluorescence microscopy was used to qualitatively compare imaged mitochondria, and their shape under different conditions. In our initial experiments, very few cells showed fluorescence, which meant that transfection occurred with very low efficiency. Transfection efficiency will be optimized before further studies are performed to observe changes in mitochondrial shape as menadione is introduced. Further studies will be established to measure ROS production using a CellROX to compare ROS generation induced by the toxicants menadione and antimycin A. Our results will be used to describe the relationship among ROS production, mitochondrial shape changes, and cell death.

Faculty Mentor(s)

Carin Thomas

Department/Program

Chemistry

Additional Mentoring Department

https://cwu.studentopportunitycenter.com/2020/04/investigating-mitochondrial-dynamics-and-cell-life-cycles-through-induced-stress/

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May 18th, 12:00 PM

Investigating Mitochondrial Dynamics and Cell Life Cycles through Induced Stress

Ellensburg

Mitochondria are organelles that produce energy rich molecules called ATP through cellular respiration. Mitochondria have been implicated in numerous disease states and could be an attractive drug target. We hypothesize that mitochondrial shape, reactive oxygen species (ROS) generation, and cell death are interrelated. During respiration, ROS are produced and can react with cell components which causes damage. Stress induced by ROS likely causes mitochondrial shape change, and in extreme cases, leads to cell death. Cell death was investigated by using a WST-8 cell viability assay on cultured mouse liver cells exposed to the oxidative stressor, menadione, which produces ROS. It was found that increased concentrations of menadione led to increased cell death. To monitor mitochondrial shape changes, a protein-based fluorescent probe was used to mark mitochondria in cells. A mitochondrial green fluorescent protein (GFP) plasmid was transfected into the mouse liver cells, so that the cells could express the mitochondrial targeted fluorescent probe. Fluorescence microscopy was used to qualitatively compare imaged mitochondria, and their shape under different conditions. In our initial experiments, very few cells showed fluorescence, which meant that transfection occurred with very low efficiency. Transfection efficiency will be optimized before further studies are performed to observe changes in mitochondrial shape as menadione is introduced. Further studies will be established to measure ROS production using a CellROX to compare ROS generation induced by the toxicants menadione and antimycin A. Our results will be used to describe the relationship among ROS production, mitochondrial shape changes, and cell death.

https://digitalcommons.cwu.edu/source/2020/COTS/33