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Updated: Jun 27, 2025
Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina
Published on: February 13, 2021
Effects of fluorescent protein tdTomato on mouse retina
Chang-Jun Zhang1, Hao Mou1, Jing Yuan1
1Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Science Key Laboratory, Beijing, 100730, China.
Abstract:
Fluorescent proteins (FPs) have been widely used to investigate cellular and molecular interactions and trace biological events in many applications. Some of the FPs have been demonstrated to cause undesirable cellular damage by light-induced ROS production in vivo or in vitro. However, it remains unknown if one of the most popular FPs, tdTomato, has similar effects in neuronal cells. In this study, we discovered that tdTomato expression led to unexpected retinal dysfunction and ultrastructural defects in the transgenic mouse retina. The retinal dysfunction mainly manifested in the reduced photopic electroretinogram (ERG) responses and decreased contrast sensitivity in visual acuity, caused by mitochondrial damages characterized with cellular redistribution, morphological modifications and molecular profiling alterations. Taken together, our findings for the first time demonstrated the retinal dysfunction and ultrastructural defects in the retinas of tdTomato-transgenic mice, calling for a more careful design and interpretation of experiments involved in FPs.
Insights
The popular tdTomato fluorescent protein caused retinal dysfunction and mitochondrial damage in transgenic mice. Researchers advise caution when using fluorescent proteins in experiments involving neuronal cells.
Area of Science:
- Neuroscience
- Cell Biology
- Biochemistry
Background:
- Fluorescent proteins (FPs) are vital tools for studying cellular processes.
- Some FPs can induce cellular damage via reactive oxygen species (ROS).
- The impact of tdTomato on neuronal cells remained uninvestigated.
Purpose of the Study:
- To determine if tdTomato expression causes adverse effects in neuronal cells.
- To investigate the potential for tdTomato to induce retinal dysfunction.
- To characterize any observed cellular or molecular alterations.
Main Methods:
- Generation of tdTomato-transgenic mice.
- Assessment of retinal function using electroretinography (ERG).
- Evaluation of visual acuity and contrast sensitivity.
- Mitochondrial morphology and molecular profiling analysis.
Main Results:
- tdTomato expression resulted in significant retinal dysfunction.
- Photopic ERG responses and contrast sensitivity were reduced.
- Mitochondrial damage, including redistribution and morphological changes, was observed.
- Molecular alterations in mitochondria were identified.
Conclusions:
- tdTomato expression induces retinal dysfunction and ultrastructural defects in mice.
- Mitochondrial damage is a key mechanism underlying tdTomato-induced toxicity.
- Careful experimental design and interpretation are necessary when using FPs like tdTomato.
