Description
Glaucoma is the leading cause of irreversible blindness. The pathophysiology is characterised by damage to retinal ganglion cells (RGCs), typically associated with raised intra-ocular pressure. An arising theory is that retinal macroglia can contribute to the progression of glaucoma when they become dysregulated. The project aims to better understand the glia and RGC interactions using RNA sequencing and protein imaging in a mouse model of glaucoma.
The silicone oil model was initiated by an intracameral injection into the anterior chamber of the mouse eye, creating a pupil block that disrupts the outflow of aqueous humor. CoppaFISH (COmbinatorial Padlock Probe Amplification Fluorescence In Situ Hybridisation), an RNA sequencing method, and IBEX (Iterative Bleaching Extends multipleXity), a multiplexed immunohistochemistry technique, were combined on the same tissue sections to classify RGC subtypes and changes to glia.
CoppaFISH and IBEX were successfully combined for RNA and protein detection, respectively, on the same mouse retinal tissue. 40 gene probes were designed and imaged, focused on RGC subsets and macroglia activation, identifying changes between silicone-oil injected and contralateral control eyes. Cell calling was attributed by genes spot counts in nuclei alongside protein labelling for the cell type and analysed with Aivia software. Correlations of subtype loss and macroglia activation was measured to explore their relationship to glaucoma progression.
CoppaFISH-IBEX has been successfully optimised to map the spatial landscape of the retina in a glaucoma-induced mouse model. This project will confirm the resiliency of RGC subtypes and relate these changes to macroglia activation, to better understand the progression of Glaucoma.
Lay Abstract
Glaucoma is the leading cause of irreversible blindness, characterised by damage of retinal ganglion cells (RGCs), cells important for vision. This damage is typically associated with raised intra-ocular pressure in glaucoma. An arising theory is that supportive cells called macroglia become dysregulated and worsen the loss of RGCs. The project aims to better understand the macroglia and RGC interactions in a mouse model of glaucoma.
The mouse model is induced by introducing a pupil block to disrupt the fluid in the eye, increasing the eye pressure. To identify changes to macroglia and RGCs we image RNA and proteins that bind to these structures. In particular, I am investigating the types of RGCs that are lost in glaucoma to find the most resistant RGC subtypes. Moreover, I will find the spatial differences of macroglia in regions with low RGCs and high RGCs to hypothesise the relationship between these cell types.
This method has been successfully optimised to map the spatial landscape of the retina in a glaucoma-induced mouse model. This project will confirm the resiliency of RGC subtypes and relate these changes to macroglia activation, to better understand the progression of Glaucoma.
| Lay Title | Characterising ganglion-cell subset loss in a model of Glaucoma |
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| Role | PhD Student |