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

eLife

Iva Ganeva¹  Michael R Wozny¹  Madeline A Lancaster¹  Patrick C Hoffmann¹  Magdalena Sutcliffe¹  Stefano L Giandomenico¹  Wanda Kukulski² 

[1] MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, United Kingdom;MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge, United Kingdom;Institute of Biochemistry and Molecular Medicine, University of Bern, Bern, Switzerland;

关键词: cryo-EM;    CLEM;    cerebral organoids;    cryo-ET;    axons;    Human;   

DOI  :  10.7554/eLife.70269

来源: eLife Sciences Publications, Ltd

【 摘 要 】

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.

【 授权许可】

CC BY