In situ cryo-electron tomography reveals the progressive biogenesis of basal bodies and cilia in mouse ependymal cells

Shanshan Ma, Luan Li, Zhixun Li, Shenjia Luo, Qi Liu, Wenjing Du, Benhua Qiu, Miao Gui (), Xueliang Zhu () and Qiang Guo ()
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Shanshan Ma: Peking University
Luan Li: Chinese Academy of Sciences
Zhixun Li: Peking University
Shenjia Luo: Peking University
Qi Liu: Zhejiang University
Wenjing Du: Peking University
Benhua Qiu: Chinese Academy of Sciences
Miao Gui: Zhejiang University
Xueliang Zhu: Chinese Academy of Sciences
Qiang Guo: Peking University

Nature Communications, 2025, vol. 16, issue 1, 1-16

Abstract: Abstract Cilia, essential organelles for cell motility and signaling, comprise an axoneme extended from the basal body (BB). The assembly process of BBs and axonemes during ciliogenesis, however, remains largely unknown due to the lack of structural information. Here, we leverage in-situ cryo-electron tomography to capture within mouse ependymal cells the dynamic processes of BB biogenesis and multiciliogenesis at various stages. This approach enables 3D visualization of the complete motile machinery, revealing the continuous microtubule-based scaffold from BBs to axonemes at sub-nanometer resolution with unprecedented structural details. Furthermore, we elucidate along BBs and cilia heterogeneous landscapes of microtubule-binding proteins underlying the establishment of structural periodicity and diverse subregions. Notably, the chronological binding patterns of microtubule-inner proteins (e.g., CEP41) correlate with the progressive assembly of ciliary beating machinery. We also resolve a substructure that borders the BB and the axoneme. Our findings provide key insights into intricate orchestrations during ciliogenesis.

Date: 2025
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DOI: 10.1038/s41467-025-61015-6

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