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Updated: Aug 1, 2025
Induction and Analysis of Oxidative Stress in Sleeping Beauty Transposon-Transfected Human Retinal Pigment Epithelial Cells
Published on: December 11, 2020
Oxidative Stress Induced Dysfunction of Protein Synthesis in 661W Mice Photoreceptor Cells
Liting Deng1, Vivek Gupta1, Morteza Abyadeh2
1Macquarie Medical School, Faculty of Medicine, Health and Human Sciences, Macquarie University, Macquarie Park, NSW 2109, Australia.
Abstract:
Photoreceptor cells are highly susceptible to oxidative-stress-induced damage due to their high metabolic rate. Oxidative stress plays a key role in driving pathological events in several different ocular diseases, which lead to retinal degeneration and ultimately blindness. A growing number of studies have been performed to understand downstream events caused by ROS induced oxidative stress in photoreceptor cells; however, the underlying mechanisms of ROS toxicity are not fully understood. To shed light on ROS induced downstream pathological events, we employed a tandem mass tag (TMT) labelling-based quantitative mass-spectrometric approach to determine proteome changes in 661W photoreceptor cells following oxidative stress induction via the application of different concentrations of H2O2 at different time points. Overall, 5920 proteins were identified and quantified, and 450 differentially expressed proteins (DEPs) were identified, which were altered in a dose and time dependent manner in all treatment groups compared to the control group. These proteins were involved in several biological pathways, including spliceosome and ribosome response, activated glutathione metabolism, decreased ECM-receptor interaction, oxidative phosphorylation, abnormally regulated lysosome, apoptosis, and ribosome biogenesis. Our results highlighted ECM receptor interaction, oxidative phosphorylation and spliceosome pathways as the major targets of oxidative stress that might mediate vascular dysfunction and cellular senescence.
Insights
Oxidative stress damages photoreceptor cells, leading to blindness. This study identified 450 proteins altered by hydrogen peroxide exposure, revealing key pathways like ECM receptor interaction and oxidative phosphorylation involved in retinal degeneration.
Area of Science:
- Ophthalmology
- Cell Biology
- Biochemistry
Background:
- Photoreceptor cells are vulnerable to oxidative stress due to high metabolic activity.
- Oxidative stress is implicated in retinal degeneration and blindness-causing ocular diseases.
- The precise mechanisms of reactive oxygen species (ROS) toxicity in photoreceptors remain unclear.
Purpose of the Study:
- To investigate proteome alterations in photoreceptor cells under oxidative stress.
- To identify key biological pathways affected by ROS-induced damage.
Main Methods:
- Utilized tandem mass tag (TMT) labeling and quantitative mass spectrometry.
- Exposed 661W photoreceptor cells to varying concentrations of hydrogen peroxide (H2O2) over different time points.
- Identified and quantified 5920 proteins, with 450 differentially expressed proteins (DEPs).
Main Results:
- Detected dose- and time-dependent changes in 450 DEPs compared to controls.
- Observed alterations in pathways including spliceosome, ribosome, glutathione metabolism, ECM-receptor interaction, oxidative phosphorylation, lysosome function, apoptosis, and ribosome biogenesis.
- Highlighted ECM receptor interaction, oxidative phosphorylation, and spliceosome pathways as significantly impacted.
Conclusions:
- Proteomic analysis reveals critical pathways affected by oxidative stress in photoreceptor cells.
- ECM receptor interaction, oxidative phosphorylation, and spliceosome pathways are major targets of oxidative stress.
- These pathways may play a role in vascular dysfunction and cellular senescence contributing to retinal degeneration.
