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首页> 外文期刊>BMC Neuroscience >3 dimensional modelling of early human brain development using optical projection tomography
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3 dimensional modelling of early human brain development using optical projection tomography

机译：使用光学投影层析成像技术对人类早期大脑进行3维建模

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Background As development proceeds the human embryo attains an ever more complex three dimensional (3D) structure. Analyzing the gene expression patterns that underlie these changes and interpreting their significance depends on identifying the anatomical structures to which they map and following these patterns in developing 3D structures over time. The difficulty of this task greatly increases as more gene expression patterns are added, particularly in organs with complex 3D structures such as the brain. Optical Projection Tomography (OPT) is a new technology which has been developed for rapidly generating digital 3D models of intact specimens. We have assessed the resolution of unstained neuronal structures within a Carnegie Stage (CS)17 OPT model and tested its use as a framework onto which anatomical structures can be defined and gene expression data mapped. Results Resolution of the OPT models was assessed by comparison of digital sections with physical sections stained, either with haematoxylin and eosin (H&E) or by immunocytochemistry for GAP43 or PAX6, to identify specific anatomical features. Despite the 3D models being of unstained tissue, peripheral nervous system structures from the trigeminal ganglion (~300 μm by ~150 μm) to the rootlets of cranial nerve XII (~20 μm in diameter) were clearly identifiable, as were structures in the developing neural tube such as the zona limitans intrathalamica (core is ~30 μm thick). Fourteen anatomical domains have been identified and visualised within the CS17 model. Two 3D gene expression domains, known to be defined by Pax6 expression in the mouse, were clearly visible when PAX6 data from 2D sections were mapped to the CS17 model. The feasibility of applying the OPT technology to all stages from CS12 to CS23, which encompasses the major period of organogenesis for the human developing central nervous system, was successfully demonstrated. Conclusion In the CS17 model considerable detail is visible within the developing nervous system at a minimum resolution of ~20 μm and 3D anatomical and gene expression domains can be defined and visualised successfully. The OPT models and accompanying technologies for manipulating them provide a powerful approach to visualising and analysing gene expression and morphology during early human brain development.
机译：背景技术随着发展的进行，人类胚胎获得了越来越复杂的三维（3D）结构。分析作为这些变化基础的基因表达模式并解释其重要性，取决于确定它们所映射的解剖结构，并随着时间的推移在开发3D结构时遵循这些模式。随着添加更多基因表达模式，尤其是在具有复杂3D结构的器官（如大脑）中，此任务的难度大大增加。光学投影层析成像（OPT）是一项新技术，已开发用于快速生成完整样本的数字3D模型。我们评估了卡内基阶段（CS）17 OPT模型中未染色的神经元结构的分辨率，并测试了其作为可定义解剖结构和映射基因表达数据的框架的用途。结果通过比较数字切片与苏木精和曙红（H＆E）染色的物理切片，或者通过针对GAP43或PAX6的免疫细胞化学，对OPT模型的分辨率进行评估，以鉴定特定的解剖特征。尽管3D模型的组织未染色，但从三叉神经节（约300μm×150μm）到颅神经XII的小根（直径约20μm）的周围神经系统结构仍清晰可辨，神经管，例如限带透明质膜（核心层厚约30μm）。已经在CS17模型中识别并显示了14个解剖区域。当将来自2D切片的PAX6数据映射到CS17模型时，两个3D基因表达域（已知由小鼠中的Pax6表达定义）清晰可见。成功证明了将OPT技术应用于从CS12到CS23的所有阶段的可行性，该阶段涵盖了人类发展中枢神经系统器官形成的主要时期。结论在CS17模型中，以至少约20μm的最小分辨率可见正在发育的神经系统内的大量细节，并且可以成功定义和可视化3D解剖结构和基因表达域。 OPT模型及其操纵技术为人类早期大脑发育过程中的可视化和分析基因表达和形态提供了一种强大的方法。

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