Function of zinc finger protein 276 during CNS myelination and remyelination as part of the oligodendrocyte transcriptional network

在脊椎动物神经系统中，髓鞘的正确形成是快速跳跃传导和维持轴突完整性的先决条件。两种类型的髓鞘形成胶质细胞，即PNS中的雪旺细胞和CNS中的少突胶质细胞负责建立这种结构，它们的分化受到不同转录调控网络的调节。在这个项目中，我将研究尚未确定的转录因子锌指蛋白276(Zfp276)在中枢神经系统髓鞘形成和再髓鞘形成过程中的作用。我在最近的转录组和芯片序列实验中确定了这个因子作为Sox10的潜在靶点，Sox10是中枢神经系统和三叉神经节中髓鞘形成的关键转录调节因子之一。利用组织学、免疫组织化学和超微结构技术，我将对携带Zfp276基因的少突胶质细胞特异性缺失的小鼠的中枢神经系统组织进行详细的表型分析。当Zfp276功能丧失或获得时，重新髓鞘形成能力的变化将使用器官型切片培养系统进行研究。我将用来自原代少突胶质细胞培养的数据来补充这些体内研究，以确定Zfp276在少突胶质细胞分化过程中的分子功能。Zfp276的表达谱和基因组占位分析将确定其在少突胶质细胞分化过程中调节其功能的调控靶点和信号通路。将我的测序数据与已发表的中枢神经系统髓鞘形成主要调控因子的RNA-Seq和ChIP-Seq数据集进行比较分析，将有助于将Zfp276整合到少突胶质细胞转录网络中，并识别上游调控因子和功能相互作用因子。

Proper formation of myelin sheaths is a prerequisite for fast saltatory conduction and maintenance of axonal integrity in the vertebrate nervous system. Two types of myelin-forming glia, namely Schwann cells in the PNS and oligodendrocytes in the CNS, are responsible for establishing this structure and their differentiation is regulated by distinct transcriptional regulatory networks. In this project I will study the role of the yet uncharacterized transcription factor zinc finger protein 276 (Zfp276) during myelination and remyelination within the CNS. I identified this factor in recent transcriptome and ChIP-Seq experiments as a potential target of Sox10, one of the key transcriptional regulators of myelination in both CNS and PNS. Using histological, immunohistochemical, and ultrastructural techniques, I will perform detailed phenotypic analyses on CNS tissue of mice that carry an oligodendrocyte-specific deletion of the Zfp276 gene. Changes in remyelination capacities upon loss or gain of Zfp276 function will be studied using the organotypic slice culture system. I will complement these in vivo studies with data from primary oligodendroglial cultures to identify molecular functions of Zfp276 during oligodendroglial differentiation. Expression profiling and analyses of genomic occupancy of Zfp276 will identify regulatory targets and signaling pathways that mediate its function during oligodendroglial differentiation. Comparative analyses of my sequencing data with published RNA-Seq and ChIP-Seq data sets for major regulators of CNS myelination, such as Sox10 and Olig2, will help to integrate Zfp276 into the oligodendroglial transcriptional network and to identify upstream regulators as well as functional interactors.