Title : Exploration of the cGMP-PKG-dependent transcriptome in photoreceptors via spatial capture technology
Abstract:
Retinitis pigmentosa (RP) refers to a group of inherited disorders that lead to the dysfunction and death of photoreceptors, which causes progressive vision loss, and there is in principle no treatment available. The cGMP-PKG system has been suggested as a disease driver in several mouse RP models, although the downstream signaling of this enzyme system has not been clarified. Here we investigate the cGMP-PKG-system with the aim to reveal its potential target(s) in the genetic level, that may have a direct detrimental effect during retinal degeneration. The so-called rd1 mouse model, with the character of fast retinal degeneration and abnormally high cGMP levels in photoreceptors, due to the mutation of gene Phosphodiesterase 6 (PDE6) as well as its normal healthy wild-type (WT) were used in this study. The mice were sacrificed at the same age of postnatal 11, with the retinas being fixed and sectioned. We previously found that Raf-1 proto-oncogene, serine/threonine kinase (RAF1) was higher expressed in the outer nuclear layer (ONL) of rd1 than WT, where photoreceptors reside and thus appears to play a vital role during retinal degeneration. We therefore immune-stained this marker on the sections and applied the spatial-based technique to explore transcriptome. The subsequent bioinformatics analysis regarding comparison of the genetic profiles specifically in ONL between these two strains will provide revealing insights in retinal degeneration, as well as details of RAF1 related mechanisms during photoreceptor death. Upon comparison, we observed a series of differential expressed genes in ONL between rd1 and WT, which are potential cGMP-PKG downstream targets and likely have a direct detrimental effect in photoreceptor degeneration. Mechanisms regarding how RAF1 functions during degeneration were also identified.
Audience Take Away:
- The differential expressed genes between rd1 and WT would be explored.
- The role of RAF1 during retinal degeneration would be described.
- Potential targets would be identified to develop novel treatments towards retinal degeneration
