A multi-modal, brain-wide atlas of astrocyte diversity across developmental stages and model species

项目摘要 星形胶质细胞影响神经元电路组装和功能，并已被证明可以响应，调节， 驱病致病。尽管如此，人们对大脑各区域星形胶质细胞的多样性知之甚少， 发展和物种。然而，最近的单细胞RNA测序研究表明， 在整个大脑中具有显著的转录组异质性。详细的分子和细胞表征 需要这些亚群来确定分子定义的星形胶质细胞亚群是否 在健康和疾病方面都有不同的功能。因此，该项目的目标是使用尖端技术， 表征星形胶质细胞亚群的转录组学，结构和功能多样性， 发育阶段和疾病相关的大脑区域在小鼠和绒猴的大脑。通过进行 我对年轻成年绒猴八个大脑区域的70，000多个星形胶质细胞核进行了详细分析， 发现星形胶质细胞之间存在显著的区域异质性，特别是在皮质和皮质下之间 地区在这个提议中，我的目标是取得三个重大进展：a）绘制星形胶质细胞区域的进化图， 发育过程中异质性，B）比较啮齿类动物和非人灵长类动物之间的这种异质性 物种，和c）表征转录组学上不同的星形胶质细胞亚型的功能特性。解决 前两个阶段，我将对胚胎，新生儿， 幼年和青春期绒猴和小鼠。为了解决第三个问题，我将使用扩展显微镜来评估 分子定义的星形胶质细胞亚型是否具有不同的形态，突触的鞘化，和/或 离子通道的纳米级结构。此外，我将图像星形胶质细胞钙动力学，考虑到 在急性脑切片中，星形胶质细胞用于调节神经元网络的信号传导过程的关键成分 以表征转录组学定义的星形胶质细胞的功能多样性 亚型这项拟议中的研究将标志着第一个跨物种，跨发展，跨地区的分子 使用一致的实验和计算方法，脑细胞的轮廓，并将创造新的 关于啮齿动物和非人类大脑区域星形胶质细胞异质性发展的知识 灵长类动物重要的是，这种详细的星形胶质细胞亚群的分子和细胞特征将 促进他们的神经发育和精神病学的基础和转化研究的精确操作 紊乱我将在冯国平博士和艾德博伊登博士的实验室进行这个项目， 马萨诸塞州理工学院科学系。冯和博伊登的实验室里 所需的设备，为拟议的项目。关于所需实验室的所有必要培训 计算技术将由高级实验室成员或通过与BCS的其他实验室合作提供 外部合作者。冯和博伊登实验室，BCS和麻省理工学院提供卓越的科学和 专业发展资源，以促进我成功过渡到博士后奖学金。

PROJECT SUMMARY Astrocytes influence neuronal circuit assembly and function and have been shown to respond, modulate, and drive disease pathogenesis. Despite this, relatively little is known about astrocyte diversity across brain regions, development, and species. However, recent single-cell RNA sequencing studies demonstrate that astrocytes have significant transcriptomic heterogeneity across the brain. Detailed molecular and cellular characterization of these subpopulations is needed to determine whether molecularly-defined astrocyte subpopulations serve distinct functions in both health and disease. Thus, the goal of this project is to use cutting-edge techniques to characterize the transcriptomic, structural, and functional diversity of astrocyte subpopulations across developmental stages and disease-relevant brain regions in the mouse and marmoset brain. By conducting a detailed analysis of over 70,000 astrocyte nuclei across eight brain regions in the young adult marmoset, I have discovered striking regional heterogeneity among astrocytes, particularly between cortical and subcortical regions. In this proposal, I aim to make three significant advances: a) map the evolution of astrocyte regional heterogeneity across development, b) compare this heterogeneity between rodent and non-human primate species, and c) characterize the functional identities of transcriptomically distinct astrocyte subtypes. To address the first two of these, I will conduct single-nucleus RNA sequencing of four brain regions in the embryo, neonate, juvenile, and adolescent marmoset and mouse. To address the third, I will use expansion microscopy to assess whether molecularly-defined astrocyte subtypes have distinct morphologies, ensheathment of synapses, and/or configuration of ion channels at the nanoscale. Additionally, I will image astrocyte calcium dynamics, considered key components of signaling processes used by astrocytes to regulate neuronal networks, in acute brain slices from both mouse and marmoset to characterize the functional diversity of transcriptomically-defined astrocyte subtypes. The proposed study will mark the first cross-species, cross-development, cross-region molecular profile of brain cells using consistent experimental and computational methodology, and will create new knowledge about the development of astrocyte heterogeneity across brain regions in rodents and non-human primates. Importantly, this detailed molecular and cellular characterization of astrocyte subpopulations will facilitate their precise manipulation for basic and translational research on neurodevelopmental and psychiatric disorders. I will carry out this project in the labs of Dr. Guoping Feng and Dr. Ed Boyden in the Brain and Cognitive Sciences Department at the Massachusetts Institute of Technology. The Feng and Boyden labs contain all required equipment for the proposed project. All necessary training regarding required laboratory and computational techniques will be provided by senior lab members or through collaboration with other labs in BCS and external collaborators. The Feng and Boyden labs, BCS, and MIT offer exceptional scientific and professional development resources to facilitate my successful transition into a postdoctoral fellowship.