Related Experiment Video
Updated: Apr 13, 2026
Primary Cell Cultures to Study the Regeneration Potential of Murine Müller Glia after MicroRNA Treatment
Published on: March 28, 2022
Age-dependent Müller glia neurogenic competence in the mouse retina
Kati Löffler1, Patrick Schäfer1,2, Manuela Völkner2
1CRTD-Center for Regenerative Therapies Dresden, Technische Universität Dresden, Dresden, 01307, Germany.
Abstract:
The mechanisms limiting neuronal regeneration in mammals and their relationship with reactive gliosis are unknown. Müller glia (MG), common to all vertebrate retinas, readily regenerate neuron loss in some species, but normally not in mammals. However, experimental stimulation of limited mammalian retina regeneration has been reported. Here, we use a mouse retina organ culture approach to investigate the MG responses at different mouse ages. We found that MG undergo defined spatio-temporal changes upon stimulation. In EGF-stimulated juvenile postmitotic retinas, most MG upregulate cell-cycle regulators (Mcm6, Pcna, Ki67, Ccnd1) within 48 h ex vivo; some also express the neurogenic factors Ascl1, Pax6, and Vsx2; up to 60% re-enter the cell cycle, some of which delaminate to divide mostly apically; and the majority cease to proliferate after stimulation. A subpopulation of MG progeny starts to express transcription factors (Ptf1a, Nr4a2) and neuronal (Calb1, Calb2, Rbfox3), but not glial, markers, indicating neurogenesis. BrdU-tracking, genetic lineage-tracing, and transgenic-reporter experiments suggest that MG reprogram to a neurogenic stage and proliferate; and that some MG progeny differentiate into neuronal-like cells, most likely amacrines, no photoreceptors; most others remain in a de-differentiated state. The mouse MG regeneration potential becomes restricted, dependent on the age of the animal, as observed by limited activation of the cell cycle and neurogenic factors. The stage-dependent analysis of mouse MG revealed similarities and differences when compared with MG-derived regeneration in fish and chicks. Therefore, the mouse retina ex vivo approach is a potential assay for understanding and overcoming the limitations of mammalian MG-derived neuronal regeneration. Postmitotic MG in mouse retina ex vivo can be stimulated to proliferate, express neurogenic factors, and generate progeny expressing neuronal or glial markers. This potential regenerative competence becomes limited with increasing mouse age.
Insights
Mammalian Müller glia (MG) can be stimulated to regenerate neurons in young mice, but this ability declines with age. This study explores the potential and limitations of mouse retinal regeneration.
Area of Science:
- Neuroscience
- Developmental Biology
- Ophthalmology
Background:
- Mammalian retinas typically lack significant neuronal regeneration.
- Müller glia (MG) are key retinal stem cells in non-mammalian vertebrates, capable of regeneration.
- Understanding limitations in mammalian MG regeneration is crucial for therapeutic strategies.
Purpose of the Study:
- To investigate the regenerative potential of mouse Müller glia (MG) in an ex vivo culture system.
- To determine how age affects MG's response to regenerative stimuli.
- To compare mammalian MG regeneration with that observed in other species.
Main Methods:
- Utilized a mouse retina organ culture model.
- Stimulated MG with epidermal growth factor (EGF).
- Employed BrdU-tracking, genetic lineage-tracing, and transgenic reporters to analyze MG behavior and progeny fate.
Main Results:
- Juvenile mouse MG, when stimulated ex vivo, upregulated cell-cycle and neurogenic factors.
- Up to 60% of MG re-entered the cell cycle, with progeny expressing neuronal markers, indicating neurogenesis.
- Regenerative potential and MG activation significantly decreased with increasing mouse age.
Conclusions:
- Postmitotic mouse MG possess latent regenerative capacity that can be experimentally induced.
- This regenerative competence is age-dependent, becoming restricted in older animals.
- The mouse retina ex vivo model is valuable for studying and potentially overcoming mammalian regenerative limitations.
