Time-Lapse Live-Cell Imaging Reveals Dual Function of Oseg4, Drosophila WDR35, in Ciliary Protein Trafficking
Nayoung Lee1, Jina Park1,4, Yong Chul Bae2, Jung Ho Lee3, Chul Hoon Kim3, and Seok Jun Moon1,* 
1Department of Oral Biology, BK21 PLUS, Yonsei University College of Dentistry, Seoul 03722, Korea, 2Department of Oral
Anatomy and Neurobiology, BK21, School of Dentistry, Kyungpook National University, Daegu 41940, Korea, 3Department of
Pharmacology, Yonsei University College of Medicine, Seoul 03722, Korea, 4Laboratory of Low Dose Risk Assessment, National
Radiation Emergency Medical Center, Korea Institute of Radiological & Medical Sciences, Seoul 01812, Korea
*Correspondence: sjmoon@yuhs.ac
Received April 2, 2018; Revised June 1, 2018; Accepted June 4, 2018.; Published online July 9, 2018.
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Cilia are highly specialized antennae-like organelles that extend from the cell surface and act as cell signaling hubs. Intraflagellar transport (IFT) is a specialized form of intracellular protein trafficking that is required for the assembly and maintenance of cilia. Because cilia are so important, mutations in several IFT components lead to human disease. Thus, clarifying the molecular functions of the IFT proteins is a high priority in cilia biology. Live imaging in various species and cellular preparations has proven to be an important technique in both the discovery of IFT and the mechanisms by which it functions. Live imaging of Drosophila cilia, however, has not yet been reported. Here, we have visualized the movement of IFT in Drosophila cilia using time-lapse live imaging for the first time. We found that NOMPB-GFP (IFT88) moves according to distinct parameters depending on the ciliary segment. NOMPB-GFP moves at a similar speed in proximal and distal cilia toward the tip (~0.45 μm/s). As it returns to the ciliary base, however, NOMPB-GFP moves at ~0.12 μm/s in distal cilia, accelerating to ~0.70 μm/s in proximal cilia. Furthermore, while live imaging NOMPB-GFP, we observed one of the IFT proteins required for retrograde movement, Oseg4 (WDR35), is also required for anterograde movement in distal cilia. We anticipate our time-lapse live imaging analysis technique in Drosophila cilia will be a good starting point for a more sophisticated analysis of IFT and its molecular mechanisms.
Keywords: Drosophila, intraflagellar transport, live imaging,
Oseg4, primary cilia

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30 June 2018 Volume 41,
Number 6, pp. 495~611

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