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Suen, K.M. orcid.org/0000-0001-9452-2528, Sheard, T.M.D. orcid.org/0000-0003-4940-3188, Lin, C.-C. et al. (3 more authors) (2023) Expansion microscopy reveals subdomains in C. elegans germ granules. Life Science Alliance, 6 (4). e202201650. ISSN 2575-1077
Abstract
Light and electron microscopy techniques have been indispensable in the identification and characterization of liquid–liquid phase separation membraneless organelles. However, for complex membraneless organelles such as the perinuclear germ granule in C. elegans, our understanding of how the intact organelle is regulated is hampered by (1) technical limitations in confocal fluorescence imaging for the simultaneous examination of multiple granule protein markers and (2) inaccessibility of electron microscopy. We take advantage of the newly developed super resolution method of expansion microscopy (ExM) and in situ staining of the whole proteome to examine the C. elegans germ granule, the P granule. We show that in small RNA pathway mutants, the P granule is smaller compared with WT animals. Furthermore, we investigate the relationship between the P granule and two other germ granules, Mutator foci and Z granule, and show that they are located within the same protein-dense regions while occupying distinct subdomains within this ultrastructure. This study will serve as an important tool in our understanding of germ granule biology and the biological role of liquid–liquid phase separation.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2023 Suen et al. This article is available under a Creative Commons License Attribution 4.0 International (https://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
Keywords: | Animals; Caenorhabditis elegans; Germ Cell Ribonucleoprotein Granules; Caenorhabditis elegans Proteins; Microscopy; Organelles |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Science (Sheffield) > School of Biosciences (Sheffield) |
Funding Information: | Funder Grant number UK RESEARCH AND INNOVATION MR/S03241X/1 INTEGRATED BIOLOGICAL IMAGING NETWORK IBIN5IJ |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 02 Mar 2023 14:51 |
Last Modified: | 23 Oct 2023 12:43 |
Published Version: | http://dx.doi.org/10.26508/lsa.202201650 |
Status: | Published |
Publisher: | Life Science Alliance, LLC |
Refereed: | Yes |
Identification Number: | 10.26508/lsa.202201650 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:196971 |
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Expansion microscopy reveals subdomains in C. elegans germ granules. (deposited 03 Mar 2023 14:31)
- Expansion microscopy reveals subdomains in C. elegans germ granules. (deposited 02 Mar 2023 14:51) [Currently Displayed]
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