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A single-molecule localization microscopy method for tissues reveals nonrandom nuclear pore distribution in Drosophila.

Accepted version
Peer-reviewed

Type

Article

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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.

Description

Keywords

Drosophila, DNA-PAINT, Lamin C, Nuclear pore complex, Super-resolution microscopy, Animals, Chromatin, Drosophila, Drosophila Proteins, Humans, Microscopy, Nuclear Envelope, Nuclear Pore

Journal Title

J Cell Sci

Conference Name

Journal ISSN

0021-9533
1477-9137

Volume Title

Publisher

The Company of Biologists
Sponsorship
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)
Wellcome Trust (203144/A/16/Z)
BBSRC (BB/V008595/1)
Jiangsu Government Scholarship for Overseas Studies, China to Jinmei Cheng
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