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Published on: August 23, 2024
Preservation of Murine Whole Eyes With Supplemented UW Cold Storage Solution: Anatomical Considerations
Nicole A Muench1, Heather M Schmitt1,2, Cassandra L Schlamp1
1Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI, USA.
Purpose:
Retinal ganglion cell (RGC) apoptosis and axon regeneration are the principal obstacles challenging the development of successful whole eye transplantation (WET). The purpose of this study was to create a neuroprotective cocktail that targets early events in the RGC intrinsic apoptotic program to stabilize RGCs in a potential donor eye.
Methods:
University of Wisconsin (UW) solution was augmented with supplements known to protect RGCs. Supplements targeted tyrosine kinase signaling, histone deacetylase activity, K+ ion efflux, macroglial stasis, and provided energy support. Modified UW (mUW) solutions with individual supplements were injected into the vitreous of enucleated mouse eyes, which were then stored in cold UW solution for 24 hours. Histopathology, immunostaining of individual retinal cell types, and analysis of cell-specific messenger RNAs (mRNAs) were used to identify supplements that were combined to create optimal mUW solution.
Results:
UW and mUW solutions reduced ocular edema and focal ischemia in globes stored in cold storage. Two major issues were noted after cold storage, including retinal detachment and reduction in glial fibrillary acidic protein staining in astrocytes. A combination of supplements resolved both these issues and performed better than the individual supplements alone. Cold storage resulted in a reduction in cell-specific mRNAs, even though it preserved the corresponding protein products.
Conclusions:
Eyes treated with optimal mUW solution exhibited preservation of retinal and cellular architecture, but did display a decrease in mRNA levels, suggesting that cold storage induced cellular stasis.
Translational Relevance:
Application of optimal mUW solution lowers an important barrier to the development of a successful whole eye transplantation procedure.
Insights
A new neuroprotective cocktail (mUW solution) stabilizes retinal ganglion cells (RGCs) for whole eye transplantation. This optimized solution preserves retinal architecture and overcomes key barriers to successful eye transplants.
Area of Science:
- Ophthalmology
- Neuroscience
- Regenerative Medicine
Background:
- Whole eye transplantation (WET) faces significant hurdles, primarily retinal ganglion cell (RGC) apoptosis and limited axon regeneration.
- Developing strategies to protect RGCs is crucial for advancing WET procedures.
Purpose of the Study:
- To create a neuroprotective cocktail targeting early RGC apoptotic pathways.
- To stabilize RGCs within donor eyes, enhancing the feasibility of WET.
Main Methods:
- Augmented University of Wisconsin (UW) solution with supplements targeting RGC survival pathways.
- Tested individual and combined supplements in enucleated mouse eyes stored for 24 hours.
- Utilized histopathology, immunostaining, and mRNA analysis to identify optimal supplement combinations.
Main Results:
- Modified UW (mUW) solutions reduced ocular edema and ischemia compared to standard UW solution.
- A combination of supplements effectively addressed retinal detachment and astrocyte changes.
- Cold storage preserved protein products but reduced cell-specific mRNA levels, indicating cellular stasis.
Conclusions:
- The optimal mUW solution preserved retinal and cellular architecture in stored eyes.
- This neuroprotective approach mitigates critical barriers to successful whole eye transplantation.
