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Related Experiment Video

Updated: Sep 9, 2025

Volumetric Imaging and Analysis of Primary Cilia in Musculoskeletal Tissue using the ARL13B-CENTRIN-2 Mouse Model

Volumetric Imaging and Analysis of Primary Cilia in Musculoskeletal Tissue using the ARL13B-CENTRIN-2 Mouse Model

Published on: March 28, 2025

Characterization of membrane structures regulating primary ciliogenesis by quantitative isotropic ultrastructure

Quanlong Lu¹, Huijie Zhao¹, Ziam Khan¹

¹Laboratory of Cellular and Developmental Signaling, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD 21702, USA.

Biorxiv : the Preprint Server for Biology

|September 2, 2025

Summary

Primary cilium assembly requires mother centriole (MC) vesicle fusion. This study reveals novel tubular membrane intermediates and their regulators, EHD1 and RAB8, crucial for ciliogenesis progression and MC protein removal.

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Last Updated: Sep 9, 2025

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Simple Detection of Primary Cilia by Immunofluorescence

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Area of Science:

Cell Biology
Membrane Trafficking
Organelle Biogenesis

Background:

Primary cilia formation depends on membrane vesicle trafficking and fusion at the mother centriole (MC).
Understanding the upstream events of axoneme growth is critical for deciphering ciliogenesis.

Purpose of the Study:

To elucidate the 3D membrane ultrastructures and proteins involved in primary cilium assembly.
To investigate the mechanisms triggering ciliogenesis progression at the mother centriole.

Main Methods:

Isotropic ultrastructure imaging
Three-dimensional quantitative analysis of membrane structures and associated proteins
Investigated protein functions (CEP164, EHD1, RAB8, IFT88, CP110/CEP97) in ciliogenesis.

Main Results:

Identified enlargement of docked vesicles at the MC as a key ciliogenesis trigger, dependent on CEP164.
Discovered novel C-shaped and toroidal membrane intermediates formed by vesicle fusion.
Demonstrated EHD1 and RAB8 orchestrate tubular intermediate formation, requiring IFT88.
Showed EHD1-mediated membrane tubulation promotes CP110/CEP97 removal from the MC cap.
Observed recruitment of ciliary gate transition zone proteins to these structures.

Conclusions:

The study redefines the architectural framework for understanding ciliogenesis.
Highlights the role of specific membrane trafficking regulators (EHD1, RAB8) and protein removal in primary cilium formation.
Emphasizes the utility of advanced imaging and analysis techniques for studying membrane trafficking and organelle biogenesis.