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Updated: Oct 11, 2026
Isolation and Culture of Primary Retinal Müller Cells from Sprague-Dawley (SD) Rats
Published on: June 17, 2025
Müller cells in the preconditioned retinal ischemic injury rat
T Nishiyama1, S Nishukawa, Hiroshi
1Department of Ophthalmology Tohoku University School of Medicine, Sendai, Japan.
Abstract:
The role of Müller cells in the preconditioned retinal ischemic injury rat was investigated. In anesthetized Sprague Dawley rats, retinal ischemia for 5 minutes constituted the preconditioning stimulus for the left eye. After 24 hours, both eyes were clamped for 60 minutes. In 30, 60, 90, and 120, minutes and 1 day, 3 days, and 7 days after ischemia, electroretinograms were recorded, and the eyeballs were enucleated. After fixation with 4% paraformaldehyde, the avidin-biotin-peroxidase technique was applied to show glutamine synthetase (GS) and glial fibrillary acidic protein (GFAP). Furthermore, for the solubilized retinas, Western blot analysis and enzyme-linked immunosorbent assay were performed to detect GS and GFAP in the extracts. Preconditioning performed 24 hours before ischemia significantly improved the recovery of the a-, and b-waves 1 day after 60 minute ischemia. In the 30, 60, 90, and 120 minutes after ischemia, the recovery of the a-wave only was observed. There was a nonsignificant trend toward greater recovery in the first 120 minutes after 60 minute ischemia, especially in the b-wave. GS immunoreactivity had no significant difference between non-preconditioned and preconditioned groups 30, 60, 90, and 120 minutes after ischemia. In 1 day after ischemia, GS immunoreactivity decreased in both groups. In 3 and 7 days after ischemia, GS immunoreactivity recovered only in the preconditioned group. The retinas at 3 and 7 days after 1 hour of ischemia showed increased GFAP immunoreactivity in the non-preconditioned group. In the preconditioned group, only slight GFAP immunoreactivity was observed. These results suggested that the mechanism of preconditioned retinal ischemia may be related to Müller cells in the retina.
Insights
Preconditioning protects the retina from ischemic injury by enhancing Müller cell recovery. This study shows preconditioning improves visual function and reduces glial scarring after retinal ischemia.
Area of Science:
- Neuroscience
- Ophthalmology
- Cell Biology
Background:
- Retinal ischemic injury can lead to significant vision loss.
- Müller cells play a crucial role in retinal homeostasis and injury response.
- Preconditioning is a potential therapeutic strategy to mitigate ischemic damage.
Purpose of the Study:
- To investigate the role of Müller cells in preconditioned retinal ischemic injury in rats.
- To evaluate the neuroprotective effects of preconditioning on retinal function and cellular markers.
Main Methods:
- Retinal ischemia was induced in Sprague Dawley rats with and without prior preconditioning.
- Electroretinograms (ERGs) were recorded to assess visual function.
- Immunohistochemistry and Western blot analysis were used to detect Müller cell markers: glutamine synthetase (GS) and glial fibrillary acidic protein (GFAP).
Main Results:
- Preconditioning significantly improved the recovery of ERG a- and b-waves 1 day after ischemia.
- GS immunoreactivity recovered in the preconditioned group by 3-7 days post-ischemia, unlike the non-preconditioned group.
- GFAP immunoreactivity, indicative of reactive gliosis, was significantly reduced in the preconditioned group compared to the non-preconditioned group.
Conclusions:
- Preconditioning enhances Müller cell recovery and reduces glial scarring following retinal ischemia.
- Müller cells are implicated in the protective mechanism of preconditioning against retinal ischemic injury.
