Department of Medical Biotechnology and Translational Medicine-Via Vanvitelli 32, Università degli Studi di Milano, 20129 Milan, Italy.
Int J Mol Sci. 2021 Jan 26;22(3):1188. doi: 10.3390/ijms22031188.
Three-dimensional (3D) reconstruction from electron microscopy (EM) datasets is a widely used tool that has improved our knowledge of synapse ultrastructure and organization in the brain. Rearrangements of synapse structure following maturation and in synaptic plasticity have been broadly described and, in many cases, the defective architecture of the synapse has been associated to functional impairments. It is therefore important, when studying brain connectivity, to map these rearrangements with the highest accuracy possible, considering the affordability of the different EM approaches to provide solid and reliable data about the structure of such a small complex. The aim of this work is to compare quantitative data from two dimensional (2D) and 3D EM of mouse hippocampal CA1 (apical dendrites), to define whether the results from the two approaches are consistent. We examined asymmetric excitatory synapses focusing on post synaptic density and dendritic spine area and volume as well as spine density, and we compared the results obtained with the two methods. The consistency between the 2D and 3D results questions the need-for many applications-of using volumetric datasets (costly and time consuming in terms of both acquisition and analysis), with respect to the more accessible measurements from 2D EM projections.
从电子显微镜(EM)数据集进行三维(3D)重建是一种广泛使用的工具,它提高了我们对大脑中突触超微结构和组织的认识。突触结构在成熟和突触可塑性方面的重排已经被广泛描述,在许多情况下,突触的缺陷结构与功能障碍有关。因此,在研究大脑连接时,考虑到不同 EM 方法的可负担性,以尽可能高的准确性来映射这些重排非常重要,以提供有关如此小而复杂结构的可靠和可靠的数据。这项工作的目的是比较来自小鼠海马 CA1(树突顶)的二维(2D)和 3D EM 的定量数据,以确定两种方法的结果是否一致。我们检查了不对称兴奋性突触,重点关注突触后密度和树突棘面积和体积以及棘密度,并比较了两种方法的结果。2D 和 3D 结果之间的一致性质疑了许多应用程序对使用体积数据集(在获取和分析方面都既昂贵又耗时)的必要性,因为 2D EM 投影提供了更易于访问的测量值。