A single-molecule localization microscopy method for tissues reveals nonrandom nuclear pore distribution in Drosophila.
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Authors
Lewków, Bohdan
Publication Date
2021-12-15Journal Title
J Cell Sci
ISSN
0021-9533
Publisher
The Company of Biologists
Type
Article
This Version
AM
Metadata
Show full item recordCitation
Cheng, J., Allgeyer, E., Richens, J., Dzafic, E., Palandri, A., Lewków, B., Sirinakis, G., & et al. (2021). A single-molecule localization microscopy method for tissues reveals nonrandom nuclear pore distribution in Drosophila.. J Cell Sci https://doi.org/10.1242/jcs.259570
Abstract
Single-molecule localization microscopy (SMLM) can provide nanoscale resolution in thin samples but has rarely been applied to tissues because of high background from out-of-focus emitters and optical aberrations. Here, we describe a line scanning microscope that provides optical sectioning for SMLM in tissues. Imaging endogenously-tagged nucleoporins and F-actin on this system using DNA- and peptide-point accumulation for imaging in nanoscale topography (PAINT) routinely gives 30 nm resolution or better at depths greater than 20 µm. This revealed that the nuclear pores are nonrandomly distributed in most Drosophila tissues, in contrast to what is seen in cultured cells. Lamin Dm0 shows a complementary localization to the nuclear pores, suggesting that it corrals the pores. Furthermore, ectopic expression of the tissue-specific Lamin C causes the nuclear pores to distribute more randomly, whereas lamin C mutants enhance nuclear pore clustering, particularly in muscle nuclei. Given that nucleoporins interact with specific chromatin domains, nuclear pore clustering could regulate local chromatin organization and contribute to the disease phenotypes caused by human lamin A/C laminopathies.
Sponsorship
Jiangsu Government Scholarship for Overseas Studies, China to Jinmei Cheng
Funder references
Wellcome Trust (092096/Z/10/Z)
Wellcome Trust (203144/Z/16/Z)
Wellcome Trust (203285/Z/16/Z)
Wellcome Trust (080007/Z/06/Z)
Wellcome Trust (080007/B/06/Z)
Wellcome Trust (109143/Z/15/Z)
Biotechnology and Biological Sciences Research Council (BB/P026486/1)
Cancer Research Uk (None)
Cancer Research UK (C6946/A24843)
Wellcome Trust (095927/B/11/Z)
Embargo Lift Date
2022-12-15
Identifiers
External DOI: https://doi.org/10.1242/jcs.259570
This record's URL: https://www.repository.cam.ac.uk/handle/1810/330699
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