Figure 5
Representative images and comparison of RNAscope data: RNAscope data confirms a loss of neuronally related genes as noted in RNA-seq results by demonstrating a decrease in the proportion of neurons expressing each neuronal probe. (A) DRG of non-diabetic individuals displayed (B) small diameter TRPV1 positive neurons and (C) larger diameter SLC17A7 positive neurons. (D) DRG of DPN individuals had decreased proportions of neurons expressing all three probes, while still containing expected neuronal subtypes such as (E) small TRPV1 positive and (F) larger SLC17A7 positive neurons. Scale bar = 100 µm. (G) Differences in proportion of neurons expressing each RNAscope between control and DPN tissue were significant at the ***p < 0.0001 level, according to the Cochran-Mantel–Haenszel Test (TRPV1 and SLC17A7), and Fisher’s Exact Test (PRDM12). Each probe was significantly downregulated, evidence of a loss of neuronally related gene expression in DPN DRG. Data is shown graphically as mean and standard error. The downregulated expression of these three neuronal markers coincides with the neuronal loss seen histologically. Although dedifferentiation could cause a loss of TRPV1 + and SLC17A7 + neurons, most neurons were still labelled with either of these two markers. White and black bars represent the control and DPN neurons expressing RNAscope, respectively.
