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Gene expression changes under cyclic mechanical stretching in rat retinal glial (Müller) cells
Xin Wang1, Jiawen Fan, Meng Zhang
1Department of Ophthalmology and Vision Sciences, Eye and ENT Hospital, Shanghai Medical College, Fudan University, Shanghai, China.
Objective:
The retina is subjected to tractional forces in various conditions. As the predominant glial element in the retina, Müller cells are active players in all forms of retinal injury and disease. In this study, we aim to identify patterns of gene expression changes induced by cyclic mechanical stretching in Müller cells.
Methods:
Rat Müller cells were seeded onto flexible bottom culture plates and subjected to a cyclic stretching regimen of 15% equibiaxial stretching for 1 and 24 h. RNA was extracted and amplified, labeled, and hybridized to rat genome microarrays. The expression profiles were analyzed using GeneSpring software, and gene ontology analysis and the Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to select, annotate, and visualize genes by function and pathway. The selected genes of interest were further validated by Quantitative Real-time PCR (qPCR).
Results:
Microarray data analysis showed that at 1 and 24 h, the expression of 532 and 991 genes in the Müller cells significantly (t-test, p<0.05) differed between the mechanically stretched and unstretched groups. Of these genes, 56 genes at 1 h and 62 genes at 24 h showed more than a twofold change in expression. Several genes related to response to stimulus (e.g., Egr2, IL6), cell proliferation (e.g., Areg, Atf3), tissue remodeling (e.g., PVR, Loxl2), and vasculogenesis (e.g., Epha2, Nrn1) were selected and validated by qPCR. KEGG pathway analysis showed significant changes in MAPK signaling at both time points.
Conclusions:
Cyclic mechanical strain induces extensive changes in the gene expression in Müller cells through multiple molecular pathways. These results indicate the complex mechanoresponsive nature of Müller cells, and they provide novel insights into possible molecular mechanisms that would account for many retinal diseases in which the retina is often subjected to mechanical forces, such as pathological myopia and proliferative vitreoretinopathy.
Insights
Mechanical stretching significantly alters gene expression in Müller cells, revealing their complex response to strain. This study highlights molecular pathways involved in retinal diseases caused by mechanical forces.
Area of Science:
- Ophthalmology
- Cell Biology
- Genomics
Background:
- Müller cells are the primary glial cells in the retina.
- These cells play a crucial role in retinal injury and disease.
- Retinal tractional forces are implicated in various ocular conditions.
Purpose of the Study:
- To investigate gene expression changes in Müller cells subjected to cyclic mechanical stretching.
- To identify specific genes and molecular pathways affected by mechanical strain.
- To understand the mechanoresponsive nature of Müller cells.
Main Methods:
- Rat Müller cells were subjected to 15% equibiaxial cyclic stretching for 1 and 24 hours.
- Gene expression profiling was performed using microarrays.
- Quantitative Real-time PCR (qPCR) was used for validation.
- Gene ontology and KEGG pathway analyses were conducted.
Main Results:
- Mechanical stretching significantly altered the expression of hundreds of genes in Müller cells at both 1 and 24 hours.
- A subset of genes related to stimulus response, cell proliferation, tissue remodeling, and vasculogenesis showed significant changes.
- The MAPK signaling pathway was identified as significantly affected by the mechanical strain.
Conclusions:
- Cyclic mechanical strain induces widespread gene expression alterations in Müller cells via multiple pathways.
- Müller cells exhibit complex mechanoresponsiveness.
- These findings offer insights into molecular mechanisms underlying retinal diseases involving mechanical forces, such as pathological myopia and proliferative vitreoretinopathy.
