Metabolic Control of Proliferation and Differentiation in Oligodendrocytes

项目摘要 髓磷脂对于适当的脑功能是至关重要的，并且其功能障碍、损伤或不适当的形成已经被发现。 据报道，在广泛的神经和精神疾病，从而敦促发现新的潜力， 治疗。该奖学金解决代谢的作用，更具体地说，葡萄糖衍生的乙酰- 辅酶A（AcCoA），调节发育髓鞘形成。实验的目的建立在一个坚实的前提上， AcCoA是一种不稳定的化合物，不能从一个隔室自由扩散到下一个隔室。总体 假设AcCoA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SLC 33 A1的水平。目的 1使用功能丧失和获得的方法来检验假设，即在第一次血糖升高期间， 出生后周有利于ACLY的核定位，从而促进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细胞增殖的基因的表达。 祖国它还假定，出生后第二周葡萄糖的短暂下降是 负责减少核ACLY，减少核AcCoA，从而有利于组蛋白脱乙酰化， OPC从增殖细胞向分化细胞的转变。该假设将使用Acly损失进行检验， 体外培养的OPC中的功能获得方法以及小鼠中的谱系特异性消融。基因组 选择的组蛋白乙酰化标记的广泛分布将使用染色质免疫沉淀进行测试。目的2 使用功能丧失和功能获得的方法来检验细胞内AcCoA合成增加的假设， 分化OL，随后将其转运至内质网（通过SLC 33 A1）对于合成 胆固醇和髓磷脂的含量。这一假设得到了以下结果的支持： Slc 33 a1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。的 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成像技术。此外，本发明还提供了一种方法， 将提供几个机会，鼓励在实验设计，数据分析，以及 书面和口头科学交流的改善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。作为有形的里程碑，这项工作预计将产生两个高质量的成果，第一， 作者手稿作为博士生，并授予机会，以获得有竞争力的博士后 任命，导致在学术科学的职业生涯。该奖学金符合申请人的长期目标， 研究脑细胞的代谢调节，NIH和NINDS的公共卫生使命是培养学术 科学家和管家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.