Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids

During brain development, axons must extend over great distances in a relatively short amount of time. How the subcellular architecture of the growing axon sustains the requirements for such rapid build-up of cellular constituents has remained elusive. Human axons have been particularly poorly acces...

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Autores principales:	Patrick C Hoffmann, Stefano L Giandomenico, Iva Ganeva, Michael R Wozny, Magdalena Sutcliffe, Madeline A Lancaster, Wanda Kukulski
Formato:	article
Lenguaje:	EN
Publicado:	eLife Sciences Publications Ltd 2021
Materias:	cryo-EM CLEM cerebral organoids cryo-ET axons Medicine R Science Q Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/02bc299a719544eaa41e48038e4ecda2
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id	oai:doaj.org-article:02bc299a719544eaa41e48038e4ecda2
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spelling	oai:doaj.org-article:02bc299a719544eaa41e48038e4ecda22021-12-01T10:55:16ZElectron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids10.7554/eLife.702692050-084Xe70269https://doaj.org/article/02bc299a719544eaa41e48038e4ecda22021-10-01T00:00:00Zhttps://elifesciences.org/articles/70269https://doaj.org/toc/2050-084XDuring brain development, axons must extend over great distances in a relatively short amount of time. How the subcellular architecture of the growing axon sustains the requirements for such rapid build-up of cellular constituents has remained elusive. Human axons have been particularly poorly accessible to imaging at high resolution in a near-native context. Here, we present a method that combines cryo-correlative light microscopy and electron tomography with human cerebral organoid technology to visualize growing axon tracts. Our data reveal a wealth of structural details on the arrangement of macromolecules, cytoskeletal components, and organelles in elongating axon shafts. In particular, the intricate shape of the endoplasmic reticulum is consistent with its role in fulfilling the high demand for lipid biosynthesis to support growth. Furthermore, the scarcity of ribosomes within the growing shaft suggests limited translational competence during expansion of this compartment. These findings establish our approach as a powerful resource for investigating the ultrastructure of defined neuronal compartments.Patrick C HoffmannStefano L GiandomenicoIva GanevaMichael R WoznyMagdalena SutcliffeMadeline A LancasterWanda KukulskieLife Sciences Publications Ltdarticlecryo-EMCLEMcerebral organoidscryo-ETaxonsMedicineRScienceQBiology (General)QH301-705.5ENeLife, Vol 10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cryo-EM CLEM cerebral organoids cryo-ET axons Medicine R Science Q Biology (General) QH301-705.5
spellingShingle	cryo-EM CLEM cerebral organoids cryo-ET axons Medicine R Science Q Biology (General) QH301-705.5 Patrick C Hoffmann Stefano L Giandomenico Iva Ganeva Michael R Wozny Magdalena Sutcliffe Madeline A Lancaster Wanda Kukulski Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
description	During brain development, axons must extend over great distances in a relatively short amount of time. How the subcellular architecture of the growing axon sustains the requirements for such rapid build-up of cellular constituents has remained elusive. Human axons have been particularly poorly accessible to imaging at high resolution in a near-native context. Here, we present a method that combines cryo-correlative light microscopy and electron tomography with human cerebral organoid technology to visualize growing axon tracts. Our data reveal a wealth of structural details on the arrangement of macromolecules, cytoskeletal components, and organelles in elongating axon shafts. In particular, the intricate shape of the endoplasmic reticulum is consistent with its role in fulfilling the high demand for lipid biosynthesis to support growth. Furthermore, the scarcity of ribosomes within the growing shaft suggests limited translational competence during expansion of this compartment. These findings establish our approach as a powerful resource for investigating the ultrastructure of defined neuronal compartments.
format	article
author	Patrick C Hoffmann Stefano L Giandomenico Iva Ganeva Michael R Wozny Magdalena Sutcliffe Madeline A Lancaster Wanda Kukulski
author_facet	Patrick C Hoffmann Stefano L Giandomenico Iva Ganeva Michael R Wozny Magdalena Sutcliffe Madeline A Lancaster Wanda Kukulski
author_sort	Patrick C Hoffmann
title	Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
title_short	Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
title_full	Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
title_fullStr	Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
title_full_unstemmed	Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
title_sort	electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids
publisher	eLife Sciences Publications Ltd
publishDate	2021
url	https://doaj.org/article/02bc299a719544eaa41e48038e4ecda2
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Electron cryo-tomography reveals the subcellular architecture of growing axons in human brain organoids

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