Oligodendrocyte cytoskeletal dynamics and transport

少突胶质细胞骨架动力学和运输

基本信息

批准号：
10932766
负责人：
Meng Meng Fu
金额：
$ 88.69万
依托单位：
NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10932766
关键词：
Action Potentials Adaptor Signaling Protein Address Adult Age Months Alzheimer&apos s Disease Animals Astrocytes Axon Behavioral Biological Assay Biophysics Brain Brain region California Career Mobility Cells Central Nervous System Chicago Code Collaborations Cytoskeleton Data Defect Development Disease Doctor of Philosophy Electron Microscopy Fright Gait Goals Golgi Apparatus Human Image Imaging Techniques Intermediate Filaments Internships Laboratories Learning Macrophage Manuscripts Messenger RNA Microfilaments Microglia Microtubule-Associated Proteins Microtubules Modeling Molecular Motor Multiple Sclerosis Multiple System Atrophy Mus Myelin Myelin Basic Proteins Myelin Proteins Myelin Sheath Myosin ATPase Myosin Type V National Heart, Lung, and Blood Institute National Institute of Mental Health National Institute of Neurological Disorders and Stroke Neurodegenerative Disorders Oligodendroglia Organelles Paper Pathogenesis Peripheral Nervous System Play Polymers Positioning Attribute Postbaccalaureate Postdoctoral Fellow Printing Process Proteins Publishing Recombinant Proteins Research Rodent Role Shapes Signal Transduction Social isolation Sorbus Stains Structure Techniques Training and Education Transgenic Organisms Translating Trees Tremor Tubulin United States National Institutes of Health Universities Untranslated Regions Visualization Work experience experimental study extracellular in vivo leukodystrophy link protein live cell imaging motor deficit mouse model myelination nervous system disorder polymerization prevent programs protein aggregation protein expression recruit remyelination response senescence therapeutic target transcriptomics

项目摘要

The laboratory closed at NINDS in July of 2023, with the PI transferring to a new position at the University of California, Berkeley. This summary represents a partial year summary. We have pursued the following goals: 1. Understanding the contribution of TPPP in neurological diseases. 2. Characterizing a mouse model in which mRNA transport in oligodendrocytes is defective. 3. Extending our research to astrocytes and microglia. 4. Pursuing collaborations. 5. Helping trainees transition to new positions. On the TPPP project, we have made significant progress and a manuscript was uploaded as a preprint in June. In human brains, we found that TPPP aggregates in MSA (multiple systems atrophy). In recombinant protein biophysical assays, we performed experiments to understand how TPPP performs its normal function to make new microtubules. Together, this project firmly establishes that TPPP is important in MSA. On the mRNA transport project, we have a working manuscript. We are specifically looking at Mbp (myelin basic protein) as the mRNA cargo. Mbp is the most abundant mRNA in oligodendrocytes and encodes a locally translated protein that is found inside the myelin sheath. Our mouse model lacks the 3 UTR region of Mbp mRNA (but contains the coding region) and was rederived by the NIMH transgenic core this spring. These animals display tremors 3 months of age and have profound motor deficits. We have now finished additional extensive behavioral analysis showing that they also have gait defects. We have finished electron microscopy analysis of both their central nervous system and peripheral nervous system myelin. We characterized Mbp mRNA transport using primary rodent oligodendrocyte cultures. Together, these data show that mRNA transport is crucial for myelination in vivo. In addition, we investigated how astrocyte make their extensive processes and branches. This project looked at 3 classes of cytoskeleton in primary ramified astrocytes: microtubules, actin, and intermediate filaments. We are putting together a short manuscript on our results. We have completed the following collaborations: 1. John Hammer (NIH, NHLBI). Dr. Hammer and I jointly recruited a research fellow, Srinivasarao Repudi, who will work on the role of different myosins in oligodendrocyte development and myelination. He discovered that myosin V actually does not affect oligodendrocyte maturation, which is different from one published observation. 2. Jeff Huang (Georgetown). We added different metabolites to primary oligodendrocyte cultures to determine whether in vivo effects observed in the Huang Lab in an in vivo remyelination model act directly on oligodendrocyte or indirectly via microglia and macrophages. We expect that a manuscript will be submitted in Winter 2023. 3. Jeff Huang (Georgetown). We added conditioned microglial media to primary cultured oligodendrocytes to determine whether senescent microglia secrete factors that prevent oligodendrocytes from differentiating and expressing key myelin proteins. A manuscript is currently in revision. We continued our collaboration with Dr. Susanne Bechstedt (McGill). Dr. Bechstedt is a long-term collaborator and we continue to work on TPPPs molecular function. Finally, several trainees pursued career transitions: 1. Hunter Richardson (postbac) started MD/PhD program at U. Chicago 2. Joseph Nowacki (postbac) started MD program at Rowan / Cooper Union. 3. William Barclay (postdoc) started detail/internship at NIH OITE (Office of Intramural Training and Education)

该实验室于2023年7月在Ninds关闭，PI转移到了加利福尼亚大学伯克利分校的新职位。该摘要代表了部分年份的摘要。我们实现了以下目标： 1。了解TPPP在神经系统疾病中的贡献。 2。表征小鼠模型，其中少突胶质细胞中的mRNA转运是有缺陷的。 3。将我们的研究扩展到星形胶质细胞和小胶质细胞。 4。进行合作。 5。帮助学员过渡到新职位。在TPPP项目中，我们取得了重大进展，手稿在6月被上传为预印本。在人的大脑中，我们发现MSA（多个系统萎缩）中的TPPP聚集体。在重组蛋白生物物理测定中，我们进行了实验，以了解TPPP如何执行其正常功能以制造新的微管。该项目共同确定了TPPP在MSA中很重要。在mRNA运输项目中，我们有一个工作手稿。我们专门将MBP（髓磷脂碱性蛋白）视为mRNA货物。 MBP是少突胶质细胞中最丰富的mRNA，并编码在髓鞘鞘内发现的局部翻译蛋白。我们的小鼠模型缺乏MBP mRNA的3 UTR区域（但包含编码区域），并在今年春季被NIMH转基因核心重新修饰。这些动物表现出3个月大的震颤，并具有深远的运动缺陷。现在，我们已经完成了其他广泛的行为分析，表明它们也具有步态缺陷。我们已经完成了它们的中枢神经系统和周围神经系统髓鞘的电子显微镜分析。我们使用原代啮齿动物少突胶质细胞培养表征MBP mRNA转运。总之，这些数据表明mRNA转运对于体内的髓鞘化至关重要。此外，我们研究了星形胶质细胞如何制作其广泛的过程和分支。该项目研究了主要分支的星形胶质细胞中的3种细胞骨架：微管，肌动蛋白和中间细丝。我们正在为我们的结果汇总一个简短的手稿。我们已经完成了以下合作： 1。JohnHammer（NIH，NHLBI）。 Hammer博士和我共同招募了一位研究员Srinivasarao Recudi，他将致力于不同肌动物在少突胶质细胞发育和髓鞘中的作用。他发现肌球蛋白V实际上不会影响少突胶质细胞的成熟，这与已发表的观察结果不同。 2。JeffHuang（乔治敦）。我们将不同的代谢物添加到原发性少突胶质细胞培养物中，以确定直接通过小胶质细胞和巨噬细胞直接对少突胶质细胞或间接地对少突胶质细胞或间接地观察到的体内实验室中观察到的体内效应。我们预计将在2023年冬季提交手稿。 3。JeffHuang（乔治敦）。我们将条件的小胶质细胞培养基添加到原发性培养的少突胶质细胞中，以确定衰老的小胶质细胞分泌因素是否可以防止少突胶质细胞区分和表达关键的髓磷脂蛋白。手稿目前正在修订中。我们继续与Susanne Bechstedt博士（McGill）合作。 Bechstedt博士是一家长期合作者，我们继续从事TPPPS分子功能。最后，几名受训者追求职业过渡： 1。HunterRichardson（Postbac）在U. Chicago开始了MD/PHD计划 2。JosephNowacki（Postbac）在Rowan / Cooper Union启动了MD计划。 3。WilliamBarclay（博士后）在NIH OETE（壁内培训和教育办公室）开始了细节/实习