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Nuclear Migration in the Drosophila Oocyte
Published on: May 13, 2021
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A single-molecule localization microscopy method for tissues reveals nonrandom nuclear pore distribution in
Jinmei Cheng1,2, Edward S Allgeyer1, Jennifer H Richens1
1The Gurdon Institute and Department of Genetics, University of Cambridge, Tennis Court Road, Cambridge CB2 1QN, UK.
Journal of Cell Science
|November 22, 2021
Summary
We developed a new microscopy technique for high-resolution imaging in tissues, revealing nuclear pore clustering in Drosophila. This finding has implications for understanding chromatin organization and laminopathies.
Area of Science:
- Cell Biology
- Microscopy
- Biophysics
Background:
- Single-molecule localization microscopy (SMLM) offers nanoscale resolution but is limited in tissue imaging due to optical challenges.
- High background noise and aberrations hinder SMLM application in complex biological tissues.
Purpose of the Study:
- To develop an optical sectioning method for SMLM in thick tissue samples.
- To investigate the spatial distribution of nuclear pores and their relationship with lamins in Drosophila tissues.
Main Methods:
- Utilized a line scanning microscope for optical sectioning in SMLM.
- Employed DNA- and peptide-point accumulation for imaging in nanoscale topography (PAINT) for high-resolution imaging.
- Visualized endogenously-tagged nucleoporins and F-actin at depths exceeding 20 µm.
Main Results:
- Achieved 30 nm resolution or better at depths >20 µm in Drosophila tissues.
- Demonstrated nonrandom nuclear pore distribution in most tissues, contrasting with cultured cells.
- Observed complementary localization of Lamin Dm0 to nuclear pores, suggesting a corralling role.
- Showed that tissue-specific Lamin C expression influences nuclear pore distribution, with mutants enhancing clustering.
Conclusions:
- Nuclear pore distribution in tissues is nonrandom and influenced by lamins.
- Nuclear pore clustering may regulate local chromatin organization.
- Findings provide insights into the pathogenesis of lamin A/C laminopathies.
