Metabolic Control of Proliferation and Differentiation in Oligodendrocytes

项目摘要 髓鞘对正常的大脑功能至关重要，它的功能障碍、损害或不适当的形成一直是 在广泛的神经和精神疾病中被报道，从而敦促发现新的潜力 治疗。这项研究涉及新陈代谢的作用，更具体地说，是葡萄糖衍生的乙酰- 辅酶A(AcCoA)，在调节发育髓鞘形成中的作用。实验的目的建立在一个坚实的前提上，即 AcCoA是一种不稳定的化合物，不能自由地从一个隔室扩散到另一个隔室。最重要的是 假设AcCoA的功能依赖于其合成酶ATP柠檬酸盐的亚细胞定位 裂解酶(ACLY)和内质网特异性AcCoA转运蛋白SLC33A1的水平。目标 1使用功能丧失和功能获得的方法来检验这样的假设，即在第一个阶段中高血糖水平 出生后一周有利于ACLY的核定位，进而促进AcCoA的合成和掺入 转化为组蛋白，从而导致有利于增殖和维持 始祖国家。它还假设，出生后第二周血糖的短暂下降是 负责降低核ACLY，减少核AcCoA从而有利于组蛋白去乙酰化和 OPC从增殖细胞向分化细胞的转变。这一假设将使用Acly Lost-And进行检验 体外培养的OPC的功能获得方法以及小鼠的谱系特异性消融。基因组 将使用染色质免疫沉淀法测试选定的组蛋白乙酰化标记的广泛分布。目标2 使用功能损失和功能获得的方法来检验细胞内AcCoA合成增加在 分化OL，然后将其运输到内质网(通过SLC33A1)是合成的关键 胆固醇和髓磷脂。这一假说得到了小鼠髓鞘增加的检测支持。 SLc33a1转基因的系统性过表达。子目标将涉及OL分化和髓鞘 通过基质辅助激光解吸/电离(MALDI)成像和电子显微镜进行显影。这个 培训计划包括学习新技能，如表观遗传学、生物信息学、 慢病毒转导、光遗传学和最新的质谱学成像技术。此外, 将提供几个机会来鼓励在实验设计、数据分析以及 提高书面和口头科学交流和指导本科生的机会。 将提供专业发展机会，并参与当地、国家和国际 会议将允许联网。作为具体的里程碑，这项工作预计将产生两个高质量的结果，首先 作者手稿作为博士生实习生，并给予机会获得有竞争力的博士后 任命，导致在学术科学的职业生涯。这项奖学金符合申请者的长期目标 研究脑细胞的代谢调节，以NIH和NINDS的公共卫生使命培养学术 科学家和管理人员在我们对大脑发育和疾病的理解方面取得了进展。

Project Summary Myelin is critical for proper brain function and its dysfunction, damage or inappropriate formation has been reported in a wide range of neurological and psychiatric disorders, thereby urging the discovery of new potential treatments. This fellowship addresses the role of metabolism, and more specifically of glucose-derived acetyl- CoA (AcCoA), in regulating developmental myelination. The experimental aims rest on the solid premise that AcCoA is an unstable compound which cannot freely diffuse from one compartment to the next. The overarching hypothesis is that AcCoA function is dependent on the subcellular localization of its synthetic enzyme ATP citrate lyase (ACLY) and on the levels of the specific AcCoA transporter to the endoplasmic reticulum, SLC33A1. Aim 1 uses loss- and gain-of-function approaches to test the hypothesis that high glucose levels during the first postnatal week favor nuclear localization of ACLY and in turn promote synthesis of AcCoA and its incorporation into histones, thereby resulting in the expression of genes that favor proliferation and the maintenance of the progenitor state. It also posits that the transient decline of glucose during the second postnatal week is responsible for decreased nuclear ACLY, decreased nuclear AcCoA thereby favoring histone deacetylation and the transition of OPC from proliferating to differentiating cells. The hypothesis will be tested using Acly loss- and gain-of-function approaches in vitro in cultured OPC as well as lineage specific ablation in mice. The genome wide distribution of select histone acetylation marks will be tested using chromatin immunoprecipitation. Aim 2 uses loss- and gain-of-function approaches to test the hypothesis that increased cytosolic AcCoA synthesis in differentiating OL, followed by its transport to the endoplasmic reticulum (via SLC33A1) is crucial for the synthesis of cholesterol and myelin lipids. This hypothesis is supported by the detection of increased myelin in mice with systemic overexpression of the Slc33a1 transgene. The subaims will address OL differentiation and myelin development by matrix assisted laser desorption/ionization (MALDI) imaging, and electron microscopy. The training plan incorporates learning of new skills, such as advanced methodology in epigenetics, bioinformatics, lentiviral transduction, optogenetics and the latest in mass spectrometry imaging technologies. In addition, several opportunities will be offered to encourage training in experimental design, data analysis, as well as improvements in written and oral scientific communication and opportunities to mentor undergraduate students. Professional development opportunities will be available and participation in local, national, and international conferences will allow networking. As tangible milestones, the work is expected to result in two high quality, first author manuscripts as a doctoral trainee, and grant the opportunity to obtain a competitive post-doctoral appointment, leading to a career in academic science. This fellowship aligns the applicant’s long-term goal of studying metabolic regulation of brain cells, with the public health mission of NIH and NINDS to foster academic scientists and steward advances in our understanding of brain development and disease.