3 dimensional modelling of early human brain development using optical projection tomography.
3 dimensional modelling of early human brain development using optical projection tomography.
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DOI：
10.1186/1471-2202-5-27
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发表时间：
2004-08-06
期刊：
影响因子：
2.4
通讯作者：
Lindsay S
中科院分区：
文献类型：
--
作者：
Kerwin J;Scott M;Sharpe J;Puelles L;Robson SC;Martínez-de-la-Torre M;Ferran JL;Feng G;Baldock R;Strachan T;Davidson D;Lindsay S
随着发育的进行，人类胚胎获得越来越复杂的三维（3D）结构。分析这些变化背后的基因表达模式并解释其意义取决于识别它们映射的解剖结构，并随着时间的推移在开发3D结构时遵循这些模式。随着更多基因表达模式的加入，这项任务的难度大大增加，特别是在具有复杂3D结构的器官中，如大脑。光学投影层析成像（OPT）是一种快速生成完整标本数字化三维模型的新技术。我们已经评估了卡内基阶段（CS）17 OPT模型内未染色的神经元结构的分辨率，并测试了其作为解剖结构可以定义和基因表达数据映射的框架的用途。通过比较数字切片与用苏木精和伊红（H&E）染色的物理切片或通过针对GAP 43或PAX 6的免疫细胞化学染色的物理切片来评估OPT模型的分辨率，以识别特定的解剖学特征。尽管3D模型是未染色的组织，但从三叉神经节（~300 μm × ~150 μm）到颅神经XII根（直径~20 μm）的周围神经系统结构清晰可辨，发育中的神经管中的结构也是如此，如丘脑内核（核心厚度~30 μm）。已在CS 17模型中识别并可视化了14个解剖结构域。当将来自2D切片的PAX 6数据映射到CS 17模型时，已知由小鼠中的Pax 6表达定义的两个3D基因表达结构域清晰可见。成功证明了将OPT技术应用于从CS 12到CS23的所有阶段的可行性，该阶段包括人类发育中枢神经系统的主要器官形成时期。在CS 17模型中，在发育中的神经系统中可以看到相当多的细节，最低分辨率为~20 μm，并且可以成功地定义和可视化3D解剖和基因表达结构域。OPT模型和相应的操作技术为可视化和分析早期人类大脑发育过程中的基因表达和形态提供了一种强大的方法。
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. 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. 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.
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影响因子：
3
作者：
Baldock, RA;Bard, JBL;Davidson, DR
通讯作者：
Davidson, DR
影响因子：
2.4
作者：
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影响因子：
2.5
作者：
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Strachan, T
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56.9
作者：
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5.7
作者：
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