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Updated: Aug 12, 2026
An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival
Published on: April 25, 2011
Single-Cell RNA Sequencing of the Primary Visual Cortex in Mice With Optic Nerve Injury
Deling Li1, Bin Zou2, Qinyuan Hu1
1State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Guangzhou, Guangdong, China.
Purpose:
Glial cells play a critical role in primary visual cortex (V1 region) damage caused by optic nerve injury, but the mechanisms driving progression of V1 region injury and glial cell heterogeneity remain poorly understood. This study aimed to investigate the damage changes in the V1 region of mice after optic nerve crush (ONC) by single-cell RNA sequencing (scRNA-seq).
Methods:
Hematoxylin and eosin (H&E) and immunofluorescence staining were used to evaluate the changes of retinal thickness, astrocytes, and microglia in the V1 region after ONC in mice. Single cell suspensions in the V1 region of mice were prepared and analyzed by scRNA-seq with Seurat, cellchat, CytoTRACE in R software. The expression of PTGDS and CRYAB was measured by qPCR, Western blot, and immunofluorescence.
Results:
After unilateral ONC, retinal thinning in both eyes and activation of astrocytes and microglia in contralateral V1 region were observed. Genes related to neuroinflammation and apoptosis in the bilateral V1 region were upregulated, and the related pathways included MAPK, TNF, and apoptosis signaling pathways. Notably, the V1 region contralateral to the ONC eye exhibited more pronounced differential gene expression, and the protein expression of neuroinflammation-related genes Ptgds and Cryab increased. We further investigated the heterogeneity and pseudotime trajectories of astrocytes and microglia, demonstrating the key branches that dominate neuroinflammation.
Conclusions:
This study generates an atlas of the V1 region of the mouse brain, highlighting the role of astrocytes and microglia in the damage changes in the V1 region after ONC, and suggesting Ptgds and Cryab as potential targets to reduce neuroinflammation.
Insights
Optic nerve crush in mice causes visual cortex (V1) damage, activating glial cells like astrocytes and microglia. This study identifies Ptgds and Cryab as potential targets to reduce associated neuroinflammation.
Area of Science:
- Neuroscience
- Immunology
- Genomics
Background:
- Glial cells are crucial in visual cortex (V1) damage after optic nerve injury.
- Mechanisms of V1 injury progression and glial cell heterogeneity are not well understood.
Purpose of the Study:
- Investigate V1 damage changes in mice post-optic nerve crush (ONC).
- Utilize single-cell RNA sequencing (scRNA-seq) to analyze V1 responses to ONC.
Main Methods:
- Evaluated retinal thickness, astrocyte, and microglia changes using H&E and immunofluorescence.
- Performed scRNA-seq on V1 single-cell suspensions using R software (Seurat, cellchat, CytoTRACE).
- Measured PTGDS and CRYAB expression via qPCR, Western blot, and immunofluorescence.
Main Results:
- ONC led to retinal thinning and contralateral V1 activation of astrocytes and microglia.
- Neuroinflammation and apoptosis-related genes/pathways (MAPK, TNF) were upregulated bilaterally.
- Contralateral V1 showed more significant differential gene expression; Ptgds and Cryab protein levels increased.
- Identified astrocyte and microglia heterogeneity and pseudotime trajectories driving neuroinflammation.
Conclusions:
- Generated a V1 region mouse brain atlas.
- Highlighted astrocyte and microglia roles in ONC-induced V1 damage.
- Suggested Ptgds and Cryab as potential therapeutic targets for neuroinflammation reduction.
