Genetic approaches to address oligodendrocyte progenitor cell diversity

摘要 少突胶质细胞发生是一个复杂的过程，需要改变转录因子的表达 少突胶质细胞前体细胞的生成、扩增和分化为成熟细胞 产生髓鞘的少突胶质细胞(OL)。这项提案的总体目标是专注于理解 在这个过程中，OPC阶段的多样性。我们的基因表达筛查发现了新的OPC标记 同源框基因Gsx1在OPC发育过程中表现出短暂的表达。在此，我们建议 开发新的小鼠模型以解决Gsx1在OPC生物学中的作用。在第一个目标中，我们将测试Gsx1是否是 在OPC中阶段特定的调节因子并生成OPC特定的条件突变体以确定是否改变Gsx1 基因功能导致OPC扩张、OL分化和/或髓鞘形成的变化。我们还将测试是否 密切相关的基因Gsx2补偿OPC中Gsx1的丢失。在第二个目标中，我们将测试是否持续 Gsx1在OL发育过程中的表达促进OPC的产生和随后的OL分化 以及使用时间控制的多西环素调节的功能增益模型的髓鞘形成。在第三个目标中，我们 将利用Gsx1谱系通过产生一本小说来揭示出生后少树发育过程中OPC的多样性 用计时器荧光蛋白检测Gsx1报告等位基因。这些研究将确定Gsx1在 OPC阶段的OL发育，并为理解复杂的调节提供了新的小鼠模型 少突胶质细胞发育过程中OPC的表达。

Abstract Oligodendrogenesis is a complex process that requires changes in transcription factor expression for the generation, expansion, and differentiation of oligodendrocyte progenitor cells (OPCs) into mature oligodendrocytes (OLs) that produce myelin. The overall goal of this proposal is to focus on understanding the diversity of the OPC stage in the process. Our gene expression screen to identity novel OPC markers revealed the homeobox gene Gsx1 exhibits temporal expression in OPCs during development. Here we propose to develop new mouse models to address the role of Gsx1 in OPC biology. In the first aim, we will test if Gsx1 is a stage specific regulator in OPCs and generate an OPC specific conditional mutant to determine if altering Gsx1 gene function causes changes in OPC expansion, OL differentiation, and/or myelination. We will also test if the closely related gene, Gsx2, compensates for the loss of Gsx1 in OPCs. In the second aim, we will test if sustained Gsx1 expression throughout OL development promotes OPC generation and subsequently OL differentiation and myelination using a temporally controlled doxycycline regulated gain-of-function model. In the third aim, we will utilize the Gsx1 lineage to reveal OPC diversity during postnatal oligodendrogenesis by generating a novel Gsx1 reporter allele using Timer Fluorescent Protein. These studies will identify the specific roles of Gsx1 during the OPC stage of OL development and provide new mouse models key for understanding the complex regulation of OPCs during oligodendrogenesis.