A single-molecule localization microscopy method for tissues reveals nonrandom nuclear pore distribution in Drosophila.
View / Open Files
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. S., Richens, J. H., 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.
Keywords
Drosophila, DNA-PAINT, Lamin C, Nuclear pore complex, Super-resolution microscopy, Animals, Chromatin, Drosophila, Drosophila Proteins, Humans, Microscopy, Nuclear Envelope, Nuclear Pore
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)
Identifiers
External DOI: https://doi.org/10.1242/jcs.259570
This record's URL: https://www.repository.cam.ac.uk/handle/1810/330699
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.
Recommended or similar items
The current recommendation prototype on the Apollo Repository will be turned off on 03 February 2023. Although the pilot has been fruitful for both parties, the service provider IKVA is focusing on horizon scanning products and so the recommender service can no longer be supported. We recognise the importance of recommender services in supporting research discovery and are evaluating offerings from other service providers. If you would like to offer feedback on this decision please contact us on: support@repository.cam.ac.uk