RNA sequencing of sciatic nerves from ZH3 and WT mice. RNA sequencing of sciatic nerves from ZH3 and WT mice

The transcriptome of sciatic nerves was investigated by RNA sequencing. We compared the transcriptome of ZH3 sciatic nerves to WT sciatic nerves in two age groups. Differentially expressed genes were considered as indicators of peripheral nerve damage in ZH3 mice. Then, we compared the sciatic nerve transcriptome of ZH3 mice chronically treated with a dimeric PrP-fusion protein (FT2Fc) to the two control groups treated with either buffer or mouse IgG. This comparison revealed specific changes induced by FT2Fc treatment. Contrary to our hypothesis, FT2Fc treatment did not alter the expression of the genes that were dysregulated in young and old ZH3 mice (as identified in the first part of the study). Overall design: Sciatic nerve mRNA profiles were obtained by deep sequencing. Number of replicates: young mice (4 months) mice: 3 WT mice, 3 ZH3 mice (both sciatic nerves from one mouse pooled into one sample). Old mice (13-15 months): 4 WT mice, 4 ZH3 mice(both sciatic nerves from one mouse pooled in each sample). Chronically treated ZH3 mice: 4 mice per treatment, each sample comprises one sciatic nerve from one mouse.

本研究通过RNA测序（RNA sequencing）探究了坐骨神经的转录组。我们针对两个年龄组，分别将ZH3小鼠的坐骨神经转录组与野生型（Wild Type，WT）小鼠坐骨神经转录组进行了对比。差异表达基因被作为ZH3小鼠外周神经损伤的指示标志物。随后，我们将长期经二聚体PrP融合蛋白（FT2Fc）处理的ZH3小鼠的坐骨神经转录组，与分别接受缓冲液或小鼠IgG处理的两个对照组进行了对比，该对比揭示了FT2Fc处理诱导的特异性转录组变化。与我们的假说相悖，FT2Fc处理并未改变年轻及老年ZH3小鼠中出现表达失调的基因的表达水平（即本研究第一部分中鉴定出的差异基因）。实验总体设计：通过深度测序获取坐骨神经mRNA表达谱。生物学重复设置如下：年轻小鼠（4月龄）：野生型小鼠3只、ZH3小鼠3只（每只小鼠的双侧坐骨神经合并为一个样本）；老年小鼠（13-15月龄）：野生型小鼠4只、ZH3小鼠4只（每只小鼠的双侧坐骨神经合并为一个样本）；长期处理的ZH3小鼠：每个处理组4只小鼠，每个样本取自1只小鼠的单侧坐骨神经。