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Updated: Aug 17, 2026
Isolation of Retinal Arterioles for Ex Vivo Cell Physiology Studies
Published on: July 14, 2018
[The experimental study of effect of pressure on rat retinal Muller cell in vitro]
Shu-Ning Li1, Jing-Hua Wang, Da-Bo Wang
1Ophthalmology Department, The Affiliated Hospital of Qingdao University, Medical College, Qingdao 266003, China. lishuningqd@hotmail.com
Objective:
To observe the changes of morphology, activity and ability of metabolizing glutamate in Muller cell and investigate the role of Muller cell in the damage induced by glaucoma.
Method:
Cultured Muller cells were divided into two groups at random, one was control group without any treatment, in the experimental group the cells were cultured under pressure of 50 mm Hg (1 mm Hg = 0.133 kPa) for 1 hour, after 3 days additional incubation, the morphology of the cells in two groups was observed by inverse phase microscope and electron microscope, the cells survival were evaluated by MTT colorimetry assay and the expression of mRNA of glutamine synthetase was semi-quantified by RT-PCR.
Result:
No obvious morphology changes were found by inverse phase microscope in the two groups, swelling mitochondrion and Vacuoles in cytoplasm were found in pressured group by electron microscope; the numbers of MTT reading and the mRNA expression in the cells under pressure were decreased, the differences was statistic significance compared with control (P < 0.05).
Conclusion:
Morphology changes were found in the cells under pressure, and the ability of metabolizing glutamate was decreased, which may be one of the reasons of the elevated level of glutamate in the vitreous of glaucoma patients.
Insights
Glaucoma pressure damages Muller cells, impairing their glutamate metabolism. This cellular dysfunction may contribute to increased glutamate levels observed in glaucoma patients.
Area of Science:
- Cell biology
- Neuroscience
- Ophthalmology
Context:
- Glaucoma is a leading cause of irreversible blindness.
- Muller cells are crucial glial cells in the retina.
- Understanding Muller cell response to pressure is vital for glaucoma research.
Purpose:
- To investigate Muller cell morphology, glutamate metabolism, and survival under simulated glaucomatous pressure.
- To elucidate the role of Muller cells in glaucoma-induced retinal damage.
Summary:
- Muller cells cultured under elevated pressure (50 mm Hg) exhibited mitochondrial swelling and cytoplasmic vacuoles.
- Cell viability (MTT assay) and glutamine synthetase mRNA expression were significantly reduced in pressured cells.
- These findings indicate impaired glutamate metabolism in Muller cells under pressure.
Impact:
- Reveals a potential mechanism for glutamate excitotoxicity in glaucoma.
- Highlights Muller cell dysfunction as a key factor in glaucoma pathogenesis.
- Provides a cellular basis for elevated vitreous glutamate levels in glaucoma.
