Using MitER for 3D analysis of mitochondrial morphology and ER contacts.
Using MitER for 3D analysis of mitochondrial morphology and ER contacts.
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DOI：
10.1016/j.crmeth.2023.100692
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发表时间：
2024-01-22
期刊：
影响因子：
--
通讯作者：
Avalos, Jose L.
中科院分区：
文献类型：
--
作者：
Kichuk, Therese;Dhamankar, Satyen;Malani, Saurabh;Hofstadter, William A.;Wegner, Scott A.;Cristea, Ileana M.;Avalos, Jose L.
我们开发了一个开源工作流程，可以对细胞器形态、分布和细胞器间接触进行定量单细胞分析，重点分析线粒体和线粒体内质网 (mito-ER) 接触位点。随着细胞器间接触的重要性得到越来越广泛的认识，对分析亚细胞结构的工具的需求也随之增加。在这里，我们描述了一个称为 MitER（发音为“mightier”）的工作流程，它允许使用动画软件 Blender 从 3D 渲染中自动计算细胞器形态、分布和细胞器间接触。然后，我们使用 MitER 来量化酿酒酵母 mito-ER 网络的变化，揭示与发酵细胞相比，呼吸细胞内的 mito-ER 接触明显更多。然后，我们演示了如何将该工作流程应用于哺乳动物系统，并用于监测延时研究中的线粒体动力学和细胞器间接触。 MitER 是用于三维细胞器渲染的开源分析工作流程 MitER 提供自动体积、表面积和细胞器间接触测量 MitER 可用于捕获细胞器形态的动态变化 线粒体形态和与近端细胞器的接触对于健康和患病细胞状态都很重要。线粒体与内质网 (ER) 接触的量化特别有价值，因为这些接触影响许多细胞过程，例如线粒体裂变和融合、线粒体自噬和钙信号传导。然而，用于此类定量分析的工具有限。当前的开源方法仅允许共定位测量，这足以估计细胞器间的接触，但缺乏提供更实质性指标的能力，例如表面积和离散接触的数量。基丘克等人。介绍 MitER，这是一个开源工作流程，可对细胞器形态、分布和细胞器间接触进行定量单细胞分析。该工作流程用于量化酿酒酵母线粒体-内质网 (mito-ER) 网络的变化，揭示与发酵细胞相比，呼吸细胞内的 mito-ER 接触显着增加。
We have developed an open-source workflow that allows for quantitative single-cell analysis of organelle morphology, distribution, and inter-organelle contacts with an emphasis on the analysis of mitochondria and mitochondria-endoplasmic reticulum (mito-ER) contact sites. As the importance of inter-organelle contacts becomes more widely recognized, there is a concomitant increase in demand for tools to analyze subcellular architecture. Here, we describe a workflow we call MitER (pronounced “mightier”), which allows for automated calculation of organelle morphology, distribution, and inter-organelle contacts from 3D renderings by employing the animation software Blender. We then use MitER to quantify the variations in the mito-ER networks of Saccharomyces cerevisiae, revealing significantly more mito-ER contacts within respiring cells compared to fermenting cells. We then demonstrate how this workflow can be applied to mammalian systems and used to monitor mitochondrial dynamics and inter-organelle contact in time-lapse studies. MitER is an open-source analysis workflow for three-dimensional organelle renderings MitER provides automated volume, surface area, and inter-organelle contact measurements MitER can be used to capture dynamic changes in organelle morphologies Mitochondrial morphology and contact with proximal organelles are important in both healthy and diseased cell states. Quantification of mitochondrial contact with the endoplasmic reticulum (ER) is particularly valuable as these contacts influence numerous cellular processes, such as mitochondrial fission and fusion, mitophagy, and calcium signaling. However, there are limited tools for such quantitative analyses. Current open-source methods solely allow for colocalization measurements, which are sufficient to estimate inter-organelle contact but lack the ability to provide more substantial metrics, such as the surface area and number of discrete contacts. Kichuk et al. introduce MitER, an open-source workflow that allows for quantitative single-cell analysis of organelle morphology, distribution, and inter-organelle contacts. The workflow is used to quantify the variations in the mitochondria-endoplasmic reticulum (mito-ER) networks of Saccharomyces cerevisiae, revealing a significant increase in mito-ER contacts within respiring cells compared to fermenting cells.
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影响因子：
9
作者：
Gómez-Suaga P;Bravo-San Pedro JM;González-Polo RA;Fuentes JM;Niso-Santano M
通讯作者：
Niso-Santano M
DOI：
10.1016/j.bbamcr.2017.05.018
发表时间：
2017-09
期刊：
Biochimica et biophysica acta. Molecular cell research
影响因子：
--
作者：
Chang CL;Chen YJ;Liou J
通讯作者：
Liou J
影响因子：
7.7
作者：
Murley A;Lackner LL;Osman C;West M;Voeltz GK;Walter P;Nunnari J
通讯作者：
Nunnari J
DOI：
10.1126/science.1207385
发表时间：
2011-10-21
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Friedman JR;Lackner LL;West M;DiBenedetto JR;Nunnari J;Voeltz GK
通讯作者：
Voeltz GK
影响因子：
4.7
作者：
Hammer, Sarah K.;Zhang, Yanfei;Avalos, Jose L.
通讯作者：
Avalos, Jose L.