Nanoimaging granule dynamics and subcellular structures in activated mast cells using soft X-ray tomography

Huan Yuan Chen, Dapi Meng Lin Chiang, Zi Jing Lin, Chia Chun Hsieh, Gung Chian Yin, I. Chun Weng, Peter Guttermann, Stephan Werner, Katja Henzler, Gerd Schneider, Lee Jene Lai, Fu-Tong Liu

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Mast cells play an important role in allergic responses. During activation, these cells undergo degranulation, a process by which various kinds of mediators stored in the granules are released. Granule homeostasis in mast cells has mainly been studied by electron microscopy (EM), where the fine structures of subcellular organelles are partially destroyed during sample preparation. Migration and fusion of granules have not been studied in detail in three dimensions (3D) in unmodified samples. Here, we utilized soft X-ray tomography (SXT) coupled with fluorescence microscopy to study the detailed structures of organelles during mast cell activation. We observed granule fission, granule fusion to plasma membranes, and small vesicles budding from granules. We also detected lipid droplets, which became larger and more numerous as mast cells were activated. We observed dramatic morphological changes of mitochondria in activated mast cells and 3D-reconstruction revealed the highly folded cristae inner membrane, features of functionally active mitochondria. We also observed giant vesicles containing granules, mitochondria, and lipid droplets, which we designated as granule-containing vesicles (GCVs) and verified their presence by EM in samples prepared by cryo-substitution, albeit with a less clear morphology. Thus, our studies using SXT provide significant insights into mast cell activation at the organelle level.

Original languageEnglish (US)
Article number34879
JournalScientific Reports
Volume6
DOIs
StatePublished - Oct 17 2016

ASJC Scopus subject areas

  • General

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    Chen, H. Y., Chiang, D. M. L., Lin, Z. J., Hsieh, C. C., Yin, G. C., Weng, I. C., Guttermann, P., Werner, S., Henzler, K., Schneider, G., Lai, L. J., & Liu, F-T. (2016). Nanoimaging granule dynamics and subcellular structures in activated mast cells using soft X-ray tomography. Scientific Reports, 6, [34879]. https://doi.org/10.1038/srep34879