应用STEM断层扫描研究应力条件下的平滑内质网形态

IF 1.9 4区 工程技术 Q3 MICROSCOPY
V. Heinz, R. Rachel, C. Ziegler
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引用次数: 0

摘要

内质网(ER)是一个高度动态的细胞器,在细胞应激反应中经历了显著的形态学改变。虽然传统的透射电子显微镜(TEM)已经为这些变化提供了有价值的见解,例如晶体内质网和内质网螺旋的形成,但在细胞背景下获得这些大型结构的全面三维(3D)信息仍然是一个挑战。为了克服这些限制,本研究引入了一种创新的双轴扫描透射电子显微镜(STEM)断层扫描技术,研究应激条件下过度表达阳离子通道多囊素-2 (PC-2)的人胚胎肾(HEK)细胞的内质网形态学。得益于高分辨率、更大的聚焦深度和更少的像差,STEM断层扫描能够对大细胞亚体积进行详细的3D重建,提供前所未有的应力诱导内质网结构视图。我们的发现揭示了晶体内质网和内质网螺旋的不同超微结构细节。晶体内质网呈管状结构,具有潜在的互联性,而内质网螺旋呈片层状组织,具有明显的膜曲率。我们观察到这些不同的光滑内质网(sER)形态在同一细胞内共同出现,但它们在空间上保持分离,这表明可能存在功能特化。此外,我们在混合形态中发现了直接的膜接触,暗示了这些结构之间的共同起源或动态关系。该研究还阐明了这些有组织的光滑内质网(OSER)结构与其他细胞器(如线粒体(MAM位点)和囊泡)的相互作用。总之,所提出的超结构见解对我们理解应力相关的内质网形态变化具有重要影响。可视化这些结构的复杂3D结构和空间关系的能力为内质网对应激的适应性反应提供了新的视角,包括在脂质和蛋白质生物合成以及细胞内通讯中的潜在作用。这些发现强调了双轴STEM断层扫描在阐明其原生细胞背景下复杂的细胞器组织和动力学方面的力量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Application of STEM tomography to investigate smooth ER morphology under stress conditions

Application of STEM tomography to investigate smooth ER morphology under stress conditions

The endoplasmic reticulum (ER) is a highly dynamic organelle that undergoes significant morphological alterations in response to cellular stress. While conventional transmission electron microscopy (TEM) has provided valuable insights into these changes, such as the formation of crystalloid-ER and ER whorls, obtaining comprehensive three-dimensional (3D) information on these large structures within their cellular context has remained a challenge. To overcome these limitations, this study introduces an innovative application of dual-axis scanning transmission electron microscopy (STEM) tomography to investigate ER morphology under stress conditions in human embryonic kidney (HEK) cells overexpressing the cation channel polycystin-2 (PC-2). Benefitting from high-resolution, increased depth-of-focus, and reduced aberrations, STEM tomography enabled the detailed 3D reconstruction of large cellular subvolumes, providing unprecedented views of stress-induced ER structures. Our findings reveal distinct ultrastructural details of both crystalloid-ER and ER whorls. Crystalloid-ER exhibited a tubular architecture with potential interconnectedness, while ER whorls displayed a lamellar organisation and distinct membrane curvature. We observed the co-occurrence of these distinct smooth ER (sER) morphotypes within the same cell, yet they remained spatially separated, suggesting potential functional specialisation. Furthermore, we identified direct membrane contacts in mixed morphotypes, hinting at a shared origin or dynamic relationship between these structures. The study also elucidated the interactions of these organised smooth ER (OSER) structures with other organelles, such as mitochondria (MAM sites) and vesicles. In summary, the presented ultra-structural insights have a significant impact on our understanding of stress-related ER morphology changes. The ability to visualise the intricate 3D architecture and spatial relationships of these structures provides novel perspectives on the ER's adaptive responses to stress, including potential roles in lipid and protein biosynthesis and intracellular communication. These findings underscore the power of dual-axis STEM tomography for elucidating complex organellar organisation and dynamics in their native cellular context.

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来源期刊
Journal of microscopy
Journal of microscopy 工程技术-显微镜技术
CiteScore
4.30
自引率
5.00%
发文量
83
审稿时长
1 months
期刊介绍: The Journal of Microscopy is the oldest journal dedicated to the science of microscopy and the only peer-reviewed publication of the Royal Microscopical Society. It publishes papers that report on the very latest developments in microscopy such as advances in microscopy techniques or novel areas of application. The Journal does not seek to publish routine applications of microscopy or specimen preparation even though the submission may otherwise have a high scientific merit. The scope covers research in the physical and biological sciences and covers imaging methods using light, electrons, X-rays and other radiations as well as atomic force and near field techniques. Interdisciplinary research is welcome. Papers pertaining to microscopy are also welcomed on optical theory, spectroscopy, novel specimen preparation and manipulation methods and image recording, processing and analysis including dynamic analysis of living specimens. Publication types include full papers, hot topic fast tracked communications and review articles. Authors considering submitting a review article should contact the editorial office first.
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