Fig. 4.

Highly cell type–specific enrichments were identified in the ONH transcriptome in FCG. a Normalized enrichment scores (NES) prioritize enrichment of different cell-specific genesets in the early FCG transcriptome. Microglia and oligodendrocyte precursor cell (OPC) and, to a lesser extent, astrocyte genesets showed upregulation in FCG, while downregulation of neuron, synaptic protein, and oligodendrocyte genesets was demonstrated. b GSEA-identified microglia- and OPC-specific gene sets exhibited statistically significant, coordinated gene upregulation in FCG (NES = 9.37 and 6.48, respectively; FDR < 0.001), whereas the neuron and myelinated oligodendrocyte gene sets were significantly downregulated (NES = − 7.80 and − 3.77, respectively; FDR < 0.001). Note that upregulation of astrocyte-specific genes is modest compared to the other ONH cell types (NES = 2.58; FDR < 0.001). Enrichment plots demonstrate cell type–specific geneset expression across the whole transcriptome. Pattern (c) and intensity (d) of immunofluorescent-labeling for the astrocyte marker GFAP in the optic nerve head were broadly similar between 10- and 12-week-old FCG and age-matched normal feline subjects (FCG: n = 3, normal: n = 3). e Normalized mRNA expression (transcript per million: TPM) of general astrocyte markers (GFAP, vimentin, aquaporin-4) in FCG from our RNA-seq dataset was not significantly increased relative to normal subjects (FDR = 0.98, 0.45, and 0.97, respectively). FCG: n = 10, normal: n = 6. Data presented as mean and SEM