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Updated: May 2, 2026
Evisceration of Mouse Vitreous and Retina for Proteomic Analyses
Published on: April 3, 2011
A multiomic study of retinal tissues in mice with direct ocular exposure to vesicants
Assylbek Zhylkibayev1, James Mobley2, Mohammad Athar3
1Wake Forest University, School of Medicine, Department of Biochemistry, Winston-Salem, NC, USA.
Abstract:
This study employed a multiomic approach to investigate retinal tissue damage following direct ocular exposure (DOE) to vesicants (VSs)-namely, nitrogen mustard (NM) and lewisite (Lew). We explored both the acute and chronic stages of retinal injury by assessing functional, structural, and molecular changes. C57BL/6 mice were used to measure scotopic and photopic electroretinograms (ERGs) and to analyze TUNEL-positive retinal cells. Global retinal proteomics was conducted to identify common and unique signaling pathways. In addition, we performed targeted metabolomic and lipidomic analyses of retinal tissue to uncover significant metabolic changes. Our results demonstrated remarkable declines in ERG amplitudes at 2 and 4 weeks post-exposure, accompanied by an increase in TUNEL+ retinal cells in response to DOE to both VSs. Our proteomic analysis revealed chronic oxidative stress, mitochondrial dysfunction, elevated RXR signaling, and increased levels of 28 proteins. Moreover, we observed a decline in the KEGG phototransduction pathways, along with the downregulation of photoreceptor-specific proteins, in response to both VSs. Consistent with the proteomic findings, targeted metabolomics identified a decline in phototransduction and steroid hormone biosynthesis, along with increases in D-amino acid and purine metabolism, as well as lysine degradation. These changes were associated with a GSSG/GSH ratio of 2.6, confirming the proteomic data on oxidative stress. Furthermore, lipidomic analysis revealed an increase in oxidative lipid levels, accompanied by a 3.4-fold increase in phosphatidylserine (PS), suggesting apoptotic cell death and a reduction in fatty acids (FAs). In conclusion, exposure to both VSs induced progressive retinal damage, altering major metabolic pathways and dysregulating lipid metabolism. Future studies should focus on identifying the responses of individual neuronal cell types to DOE to VSs to develop cell-specific countermeasures.
Insights
Direct ocular exposure to vesicants like nitrogen mustard and lewisite causes progressive retinal damage. This multiomic study reveals significant functional, structural, and molecular changes, including oxidative stress and altered metabolic pathways.
Area of Science:
- Ophthalmology
- Toxicology
- Molecular Biology
Background:
- Vesicant exposure, such as nitrogen mustard and lewisite, poses a significant threat to ocular health.
- Understanding the long-term effects of vesicant exposure on retinal tissue is crucial for developing effective countermeasures.
Purpose of the Study:
- To investigate the multiomic changes in retinal tissue following direct ocular exposure to nitrogen mustard and lewisite.
- To assess both acute and chronic stages of retinal injury, including functional, structural, and molecular alterations.
Main Methods:
- Multiomic analysis including proteomics, metabolomics, and lipidomics in C57BL/6 mice.
- Assessment of electroretinograms (ERGs) and TUNEL-positive retinal cells.
- Quantification of oxidative stress markers (GSSG/GSH ratio) and lipid profiles.
Main Results:
- Significant declines in ERG amplitudes and increased TUNEL-positive cells were observed post-exposure.
- Proteomic analysis revealed chronic oxidative stress, mitochondrial dysfunction, and altered RXR signaling.
- Metabolomic and lipidomic analyses showed dysregulation in phototransduction, steroid hormone biosynthesis, and increased oxidative lipids, indicative of apoptosis.
Conclusions:
- Vesicant exposure induces progressive retinal damage, characterized by functional deficits and molecular alterations.
- Key metabolic and lipid pathways are significantly disrupted, highlighting the systemic impact on retinal tissue.
- Further research into cell-specific responses is needed to develop targeted therapeutic strategies.
