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A Protocol to Evaluate and Quantify Retinal Pigmented Epithelium Pathologies in Mouse Models of Age-Related Macular Degeneration
Published on: March 10, 2023
A Protocol to Evaluate and Quantify Retinal Pigmented Epithelium Pathologies in Mouse Models of Age-Related Macular
Michael Landowski1, Samuel Grindel2, Ying Hao3
1Department of Medical Genetics, University of Wisconsin-Madison; McPherson Eye Research Institute, University of Wisconsin-Madison.
Abstract:
Age-related macular degeneration (AMD) is a debilitating retinal disorder in aging populations. It is widely believed that dysfunction of the retinal pigmented epithelium (RPE) is a key pathobiological event in AMD. To understand the mechanisms that lead to RPE dysfunction, mouse models can be utilized by researchers. It has been established by previous studies that mice can develop RPE pathologies, some of which are observed in the eyes of individuals diagnosed with AMD. Here, we describe a phenotyping protocol to assess RPE pathologies in mice. This protocol includes the preparation and evaluation of retinal cross-sections using light microscopy and transmission electron microscopy, as well as that of RPE flat mounts by confocal microscopy. We detail the common types of murine RPE pathologies observed by these techniques and ways to quantify them through unbiased methods for statistical testing. As proof of concept, we use this RPE phenotyping protocol to quantify the RPE pathologies observed in mice overexpressing transmembrane protein 135 (Tmem135) and aged wild-type C57BL/6J mice. The main goal of this protocol is to present standard RPE phenotyping methods with unbiased quantitative assessments for scientists using mouse models of AMD.
Insights
This study presents a standardized protocol for assessing retinal pigmented epithelium (RPE) pathologies in mouse models of age-related macular degeneration (AMD). It offers unbiased quantification methods crucial for understanding AMD mechanisms.
Area of Science:
- Ophthalmology
- Cell Biology
- Genetics
Background:
- Age-related macular degeneration (AMD) is a leading cause of vision loss in aging populations.
- Retinal pigmented epithelium (RPE) dysfunction is a critical factor in AMD pathogenesis.
- Mouse models are valuable tools for studying AMD, exhibiting RPE pathologies similar to human patients.
Purpose of the Study:
- To describe a comprehensive phenotyping protocol for evaluating RPE pathologies in mice.
- To provide standardized, unbiased methods for quantifying RPE abnormalities in research settings.
- To facilitate the study of AMD mechanisms using genetically modified and aged mouse models.
Main Methods:
- Utilizes light microscopy and transmission electron microscopy for retinal cross-section analysis.
- Employs confocal microscopy for evaluating RPE flat mounts.
- Details common murine RPE pathologies and their unbiased quantification.
Main Results:
- Demonstrates the application of the protocol in mice overexpressing TMEM135 and aged wild-type mice.
- Provides quantitative data on RPE pathologies in these models.
- Establishes a reliable method for assessing RPE health in mouse models.
Conclusions:
- The described protocol offers a robust framework for RPE phenotyping in mouse models of AMD.
- Standardized, quantitative assessment is essential for advancing AMD research.
- This methodology supports the investigation of genetic and age-related factors in RPE dysfunction.
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