Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy

The synaptonemal complex (SC) is a meiosis-specific nuclear multiprotein complex that is essential for proper synapsis, recombination and segregation of homologous chromosomes. We combined structured illumination microscopy (SIM) with different expansion microscopy (ExM) protocols including U-ExM, p...

Descripción completa

Detalles Bibliográficos
Autores principales: Zwettler, Fabian U., Spindler, Marie-Christin, Reinhard, Sebastian, Klein, Teresa, Kurz, Andreas, Benavente, Ricardo, Sauer, Markus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7320163/
https://www.ncbi.nlm.nih.gov/pubmed/32591508
http://dx.doi.org/10.1038/s41467-020-17017-7
_version_ 1783551182709456896
author Zwettler, Fabian U.
Spindler, Marie-Christin
Reinhard, Sebastian
Klein, Teresa
Kurz, Andreas
Benavente, Ricardo
Sauer, Markus
author_facet Zwettler, Fabian U.
Spindler, Marie-Christin
Reinhard, Sebastian
Klein, Teresa
Kurz, Andreas
Benavente, Ricardo
Sauer, Markus
author_sort Zwettler, Fabian U.
collection PubMed
description The synaptonemal complex (SC) is a meiosis-specific nuclear multiprotein complex that is essential for proper synapsis, recombination and segregation of homologous chromosomes. We combined structured illumination microscopy (SIM) with different expansion microscopy (ExM) protocols including U-ExM, proExM, and magnified analysis of the proteome (MAP) to investigate the molecular organization of the SC. Comparison with structural data obtained by single-molecule localization microscopy of unexpanded SCs allowed us to investigate ultrastructure preservation of expanded SCs. For image analysis, we developed an automatic image processing software that enabled unbiased comparison of structural properties pre- and post-expansion. Here, MAP-SIM provided the best results and enabled reliable three-color super-resolution microscopy of the SCs of a whole set of chromosomes in a spermatocyte with 20–30 nm spatial resolution. Our data demonstrate that post-expansion labeling by MAP-SIM improves immunolabeling efficiency and allowed us thus to unravel previously hidden details of the molecular organization of SCs.
format Online
Article
Text
id pubmed-7320163
institution National Center for Biotechnology Information
language English
publishDate 2020
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-73201632020-06-30 Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy Zwettler, Fabian U. Spindler, Marie-Christin Reinhard, Sebastian Klein, Teresa Kurz, Andreas Benavente, Ricardo Sauer, Markus Nat Commun Article The synaptonemal complex (SC) is a meiosis-specific nuclear multiprotein complex that is essential for proper synapsis, recombination and segregation of homologous chromosomes. We combined structured illumination microscopy (SIM) with different expansion microscopy (ExM) protocols including U-ExM, proExM, and magnified analysis of the proteome (MAP) to investigate the molecular organization of the SC. Comparison with structural data obtained by single-molecule localization microscopy of unexpanded SCs allowed us to investigate ultrastructure preservation of expanded SCs. For image analysis, we developed an automatic image processing software that enabled unbiased comparison of structural properties pre- and post-expansion. Here, MAP-SIM provided the best results and enabled reliable three-color super-resolution microscopy of the SCs of a whole set of chromosomes in a spermatocyte with 20–30 nm spatial resolution. Our data demonstrate that post-expansion labeling by MAP-SIM improves immunolabeling efficiency and allowed us thus to unravel previously hidden details of the molecular organization of SCs. Nature Publishing Group UK 2020-06-26 /pmc/articles/PMC7320163/ /pubmed/32591508 http://dx.doi.org/10.1038/s41467-020-17017-7 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Zwettler, Fabian U.
Spindler, Marie-Christin
Reinhard, Sebastian
Klein, Teresa
Kurz, Andreas
Benavente, Ricardo
Sauer, Markus
Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
title Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
title_full Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
title_fullStr Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
title_full_unstemmed Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
title_short Tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
title_sort tracking down the molecular architecture of the synaptonemal complex by expansion microscopy
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7320163/
https://www.ncbi.nlm.nih.gov/pubmed/32591508
http://dx.doi.org/10.1038/s41467-020-17017-7
work_keys_str_mv AT zwettlerfabianu trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy
AT spindlermariechristin trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy
AT reinhardsebastian trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy
AT kleinteresa trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy
AT kurzandreas trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy
AT benaventericardo trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy
AT sauermarkus trackingdownthemoleculararchitectureofthesynaptonemalcomplexbyexpansionmicroscopy

Ejemplares similares