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Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale
The axonal cytoskeleton is organized in a highly periodic structure, the membrane-associated periodic skeleton (MPS), which is essential to maintain the structure and function of the axon. Here, we use stimulated emission depletion microscopy of primary rat cortical neurons in microfluidic chambers...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9399097/ https://www.ncbi.nlm.nih.gov/pubmed/35999340 http://dx.doi.org/10.1038/s41598-022-18562-5 |
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author | Hofmann, Max Biller, Lucas Michel, Uwe Bähr, Mathias Koch, Jan Christoph |
author_facet | Hofmann, Max Biller, Lucas Michel, Uwe Bähr, Mathias Koch, Jan Christoph |
author_sort | Hofmann, Max |
collection | PubMed |
description | The axonal cytoskeleton is organized in a highly periodic structure, the membrane-associated periodic skeleton (MPS), which is essential to maintain the structure and function of the axon. Here, we use stimulated emission depletion microscopy of primary rat cortical neurons in microfluidic chambers to analyze the temporal and spatial sequence of MPS formation at the distal end of growing axons and during regeneration after axotomy. We demonstrate that the MPS does not extend continuously into the growing axon but develops from patches of periodic βII-spectrin arrangements that grow and coalesce into a continuous scaffold. We estimate that the underlying sequence of assembly, elongation, and subsequent coalescence of periodic βII-spectrin patches takes around 15 h. Strikingly, we find that development of the MPS occurs faster in regenerating axons after axotomy and note marked differences in the morphology of the growth cone and adjacent axonal regions between regenerating and unlesioned axons. Moreover, we find that inhibition of the spectrin-cleaving enzyme calpain accelerates MPS formation in regenerating axons and increases the number of regenerating axons after axotomy. Taken together, we provide here a detailed nanoscale analysis of MPS development in growing axons. |
format | Online Article Text |
id | pubmed-9399097 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-93990972022-08-25 Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale Hofmann, Max Biller, Lucas Michel, Uwe Bähr, Mathias Koch, Jan Christoph Sci Rep Article The axonal cytoskeleton is organized in a highly periodic structure, the membrane-associated periodic skeleton (MPS), which is essential to maintain the structure and function of the axon. Here, we use stimulated emission depletion microscopy of primary rat cortical neurons in microfluidic chambers to analyze the temporal and spatial sequence of MPS formation at the distal end of growing axons and during regeneration after axotomy. We demonstrate that the MPS does not extend continuously into the growing axon but develops from patches of periodic βII-spectrin arrangements that grow and coalesce into a continuous scaffold. We estimate that the underlying sequence of assembly, elongation, and subsequent coalescence of periodic βII-spectrin patches takes around 15 h. Strikingly, we find that development of the MPS occurs faster in regenerating axons after axotomy and note marked differences in the morphology of the growth cone and adjacent axonal regions between regenerating and unlesioned axons. Moreover, we find that inhibition of the spectrin-cleaving enzyme calpain accelerates MPS formation in regenerating axons and increases the number of regenerating axons after axotomy. Taken together, we provide here a detailed nanoscale analysis of MPS development in growing axons. Nature Publishing Group UK 2022-08-23 /pmc/articles/PMC9399097/ /pubmed/35999340 http://dx.doi.org/10.1038/s41598-022-18562-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Hofmann, Max Biller, Lucas Michel, Uwe Bähr, Mathias Koch, Jan Christoph Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
title | Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
title_full | Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
title_fullStr | Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
title_full_unstemmed | Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
title_short | Cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
title_sort | cytoskeletal assembly in axonal outgrowth and regeneration analyzed on the nanoscale |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9399097/ https://www.ncbi.nlm.nih.gov/pubmed/35999340 http://dx.doi.org/10.1038/s41598-022-18562-5 |
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