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

Published version
Peer-reviewed

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Abstract

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.

Description

Funder: Natural Sciences and Engineering Research Council of Canada; FundRef: http://dx.doi.org/10.13039/501100000038

Keywords

CLEM, axons, cell biology, cerebral organoids, cryo-EM, cryo-ET, human, Axons, Brain, Cryoelectron Microscopy, Electron Microscope Tomography, HeLa Cells, Humans, Macromolecular Substances, Microscopy, Microscopy, Fluorescence, Organoids

Journal Title

Elife

Conference Name

Journal ISSN

2050-084X
2050-084X

Volume Title

10

Publisher

eLife Sciences Publications, Ltd
Sponsorship
Medical Research Council (MC_UP_1201/9)
European Research Council (ERC STG 757710)
Medical Research Council (MC_UP_1201/8)