Role of Rnf40-dependent histone monoubiquitination in peripheral nervous system myelination [RNA-Seq]

To address the role of monoubiquitination of histone H2B at lysine 120 (H2Bub1), we inactivated the responsible E3 ligase by deleting Rnf40 in developing Schwann cells. Rnf40 becomes important for proper Schwann cell development shortly before and during differentiation. Its absence in Schwann cells leads to a peripheral neuropathy. RNA-Seq studies were performed on sciatic nerves from control mice and mice with a Schwann cell-specific deletion of Rnf40 to analyze changes in gene expression. These revealed substantial changes in the expression of genes associated with myelination, lipid metabolism and cell adhesion. We generated mRNA profiles of sciatic nerves from 14-day old control mice (ctrl; carrying floxed Rnf40 alleles) and mice with a Schwann cell-specific Rnf40 deletion (cko; carrying Rnf40 alleles in combination with a Dhh::Cre transgene) using the Illumina HiSeq 2500 platform.

为明确组蛋白H2B赖氨酸120位点单泛素化（H2Bub1）的生物学功能，我们通过在发育中的施万细胞中敲除Rnf40，以失活其对应的E3泛素连接酶。Rnf40在施万细胞分化前夕及分化进程中，对其正常发育至关重要。施万细胞中Rnf40的缺失会引发周围神经病。为分析基因表达变化情况，我们对对照组小鼠及施万细胞特异性敲除Rnf40的小鼠的坐骨神经开展了RNA测序（RNA-Seq）研究。结果显示，与髓鞘形成、脂质代谢及细胞黏附相关的基因表达发生了显著改变。本研究依托Illumina HiSeq 2500测序平台，对14日龄对照组小鼠（标注为ctrl，携带floxed Rnf40等位基因）及施万细胞特异性Rnf40敲除小鼠（标注为cko，携带Rnf40等位基因并结合Dhh::Cre转基因）的坐骨神经进行了mRNA表达谱分析。