Friend or foe? Investigating the role of microglia in chronic photoreceptor degeneration using zebrafish

点击翻译按钮获取中文摘要

Microglia play key roles in chronic degenerations in the eye, but whether they perform neuroprotective or neurotoxic functions is still unclear. To clarify these roles, I am studying a zebrafish degeneration mutant that undergoes photoreceptor degeneration throughout its lifespan, but that also exhibits associated regeneration around 4 weeks old. I collected fluorescent protein-expressing microglia from mutant and sibling zebrafish eyes at 5 days old (onset of degeneration) and 4 weeks old (start of associated regeneration) using flow cytometry. The 10x Chromium platform was then used to generate libraries for sequencing, and the samples were sequenced.Once the sequencing data was obtained, I performed quality control and clustering analysis using R packages such as Seurat to investigate the composition of the microglial population at 5 days old and 4 weeks old. I performed several iterations of the analysis by changing parameters such as clustering resolution, to ensure that interpretations were robust. Through differential expression testing, exploration of the literature, and GO enrichment analysis, I made candidate lists of genes that I hypothesize are controlling microglial function at the onset of degeneration and start of regeneration. To visualize the expression of these candidate genes in the mutant and sibling retina, I began to prepare for in vivo analysis by standardizing protocols and generating probes for in situ hybridization.