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

髓鞘的正常形成是脊椎动物神经系统快速跳跃传导和维持轴突完整性的先决条件。两种类型的髓磷脂形成胶质细胞，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，它们的分化受不同的转录调节网络的调节。在这个项目中，我将研究尚未表征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中的作用。我在最近的转录组和ChIP-Seq实验中发现这个因子是Sox10的潜在靶标，Sox10是中枢神经系统和PNS中髓鞘形成的关键转录调节因子之一。使用组织学、免疫组织化学和超微结构技术，我将对携带少突胶质细胞特异性缺失Zfp276基因的小鼠中枢神经系统组织进行详细的表型分析。在丧失或获得Zfp276功能时再髓鞘再生能力的变化将使用有机型切片培养系统进行研究。我将用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用分析将确定在少突胶质分化过程中介导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RNA-Seq和ChIP-Seq数据集进行比较分析，这些数据集针对中枢神经系统髓鞘形成的主要调节因子，如Sox10和Olig2，将有助于将Zfp276整合到少枝胶质转录网络中，并识别上游调节因子以及功能相互作用因子。

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.