Project 2

摘要(利希特曼) 突触连接的数量或模式的异常(“连接性病变”)很可能是基础 神经发育和精神障碍，如自闭症谱系障碍和精神分裂症。但 寻找大脑回路中的病理结构基元可能需要足够的分辨率来分析每个突触 并确定将数千个突触前和突触后神经元连接在一起的网络。如此高的分辨率 体积结构分析可以用连续切片电子显微镜来实现，然而这一过程 一直是极其缓慢和劳动密集型的，阻碍了样品的比较和数量的分析 包含多个神经元的。为了解决这些问题，我们设计了一台基本上自动化的计算机， 技术密集型管道，以克服迄今阻碍发现 大脑回路的结构异常。我们建议使用这套自动化显微镜和分析 从小鼠、绒猴和人(大脑)重建岛状或前额叶大脑皮质回路的工具 有机化合物和实际大脑皮层)。AIMS旨在提供突触和电路级信息 关于我们CONTE团队其他成员正在研究的分子微扰的影响。这其中的每一个 扰动与人类神经精神和神经发育障碍有关。总而言之，10 实验脑组织和它们的对照将被分析。我们将全面清查结构性库存 决定大脑皮层厚度的其他因素包括：神经元的数量和类型； 和突触的类型；每个突触的突触小泡数量，每个突触的线粒体数量和密度 以及活动区的大小、树突棘的数量、密度和大小。此外，我们将逐项列出 胶质细胞的类型，他们的流行，并寻找不同的是胶质细胞的结构。所有这些方法都有 之前被我们使用过。最后，我们将使用种子细胞方法重建电路，我们还拥有 以前开发的。我们希望提供这种详细的突触和连接信息 因为许多异常组织和对照组织将有助于集中研究疾病中身体异常的部位 到目前为止，这方面几乎没有结构性支撑。

ABSTRACT (Lichtman) It is likely that abnormalities in the number or pattern of synaptic connections (“connectopathies”) underlie neurodevelopmental and psychiatric disorders such as autism spectrum disorder and schizophrenia. But finding pathological structural motifs in brain circuitry may require sufficient resolution to analyze each synapse and identify the networks linking thousands of pre- and postsynaptic neurons together. Such high resolution volumetric structural analysis can be achieved with serial section electron microscopy, however this process has been extremely slow and labor intensive preventing the comparison of samples and analysis of volumes containing multiple neurons. To solve these problems we have devised a largely automated computer and technology intensive pipeline to overcome the principal obstacles that have to date prevented discovering structural abnormalities in brain circuits. We propose to use this suite of automated microscopy and analysis tools to reconstruct circuits in insular or prefrontal cerebral cortex from mouse, marmoset and human (cerebral organoids and actual cerebral cortex). The aims are designed to provide synaptic and circuit level information about the effects of molecular perturbations being studied by other members of our Conte team. Each of these perturbations is associated with human neuropsychiatric and neurodevelopmental disorders. In all, 10 experimental brain tissues and their controls will be analyzed. We will take a structural inventory for a full thickness of cerebral cortex determining among other things: the number and types of neurons; the number and types of synapses; the synaptic vesicle numbers per synapse, mitochondrial numbers and densities per synapse; and sizes of active zones, dendritic spine number, density and size. In addition we will itemize the glial cell types, their prevalence, and look for differences is glial cell structure. All of these approaches have been used previously by us. Finally we will reconstruct circuits using a seed cell approach that we have also previously developed. It is our hope that providing this kind of detailed synaptic and connectional information for many abnormal and control tissue will help focus research on the sites of physical abnormality in diseases that to date have little in the way structural underpinnings.