DNA修复基因ogg1调节脑发育的易损性及其分子机制

It has been demonstrated that oxidative stress can affect brain development, and DNA repair gene ogg1, an enzyme for oxidative damage repair, plays a critical role in brain development. However, the temporal-spatial expression and precise localization of ogg1 remain elusive. The impact of ogg1 deficiency on brain development is lack of deep investigation on molecular mechanisms. This project focuses on a series of scientific questions: what’s the expression spectrum and dynamic localization of ogg1 in the developing brain? How does ogg1 modulate the vulnerability of the developing brain? And what are the underlying molecular mechanisms? The study will illuminate the temporal-spatial expression pattern by real time RT-PCR and whole mount in situ hybridization during different developmental stages in zebrafish. The fluorescence labeled ogg1 transgenic lines will be generated to trace the ogg1 positive cells migration in the brain. We will construct the transgenic lines with fluorescence labeled neural stem cell in order to explore the effects of ogg1 deficiency on neural stem cell physiology, cooperating with FAC sorting and morpholino knockdown technologies. The genomic deep sequencing technology is employed to look into the molecular mechanisms. The index differences such as proliferation, apoptosis and the integration of synapses of neural cells in the brain between control and knockdown groups will be constitutively analyzed. We will use gene chip to screen the downstream effective molecular and confirm the associations by using clinical samples. By the execution of this project, we can preliminarily reveal the roles of DNA repair gene ogg1 in the brain developmental physiology and pathological phenotypes.

已证实,氧化应激影响神经发育，DNA修复基因ogg1作为氧化损伤的修复酶，对脑发育具重要作用。但，ogg1的时空表达和精细定位至今未明， ogg1缺失对于脑发育的影响也鲜有深入的机制探讨。本课题围绕“ogg1在脑发育中表达图谱和动态定位如何？如何调控脑发育的易损性？分子机制如何”这一系列科学问题，通过不同发育阶段斑马鱼的实时RT-PCR和全胚胎原位杂交明确ogg1的时空表达；构建荧光标记ogg1的转基因鱼动态追踪ogg1阳性细胞的脑内迁移；构建荧光标记神经干细胞转基因鱼系，结合流式细胞分选技术和吗啉代（MO）基因敲除，探讨ogg1缺失对神经干细胞生理功能的影响，并运用基因组深度测序探讨其分子机制；比较敲除组及对照，脑内神经细胞的增殖、凋亡、突触完整性等指标的差异，芯片分析筛检下游效应分子，并在临床病例中验证相关性。本研究可初步揭示DNA修复基因ogg1在脑发育的生理过程及病理表型中的作用。

氧化应激影响脑发育，DNA 修复基因ogg1 作为氧化损伤的修复酶，对脑发育具重要作用。课题实施通过斑马鱼神经发育模型、小鼠神经损伤模型、结合临床颅脑发育异常病例，探讨脑发育异常的病因学机制，并在临床病例中，验证和发现脑发育的关键基因（ogg1 等）和调控网络。.项目执行期共发表通讯作者SCI 论文8 篇，申请专利1项。参加国内外学术交流9人次。项目实施过程顺利，完成预期目标。.