期刊
ELIFE
卷 10, 期 -, 页码 -出版社
eLIFE SCIENCES PUBL LTD
DOI: 10.7554/eLife.70269
关键词
cryo-EM; CLEM; cerebral organoids; cryo-ET; axons; Human
类别
资金
- Medical Research Council [MC_UP_1201/9, MC_UP_1201/8]
- European Research Council [ERC STG 757710]
- Natural Sciences and Engineering Research Council of Canada
This study presents a new method combining cryo-correlative light microscopy and electron tomography with human cerebral organoid technology to visualize axon growth. The data reveal structural details on macromolecule arrangement, cytoskeletal components, and organelles, highlighting the role of the endoplasmic reticulum in lipid biosynthesis for growth support. The scarcity of ribosomes within the expanding shaft suggests limited translational competence during expansion.
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.
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