A transgenic zebrafish for in vivo visualization of cilia
Hongyu Zhang,
Zhuoya Huang,
Liuliu LV,
Yuye Xin,
Qian Wang,
Feng Li,
Lina Dong,
Changxin Wu,
Philip W. Ingham,
Zhonghua Zhao
Affiliations
Hongyu Zhang
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Zhuoya Huang
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Liuliu LV
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Yuye Xin
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Qian Wang
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Feng Li
Department of Molecular Biology, Shanxi Cancer Hospital, Affiliated Cancer Hospital of Shanxi Medical University, Taiyuan 030013, People's Republic of China
Lina Dong
Central Laboratory, Shanxi Provincial People's Hospital, Affiliate of Shanxi Medical University, Taiyuan 030012, People's Republic of China
Changxin Wu
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Philip W. Ingham
LKC Medicine School, Nanyang Technological University, 639798, Singapore
Zhonghua Zhao
Institute of Biomedical Sciences, 1331 Local Bio-Resources and Health Industry Collaborative Innovation Center of Shanxi Province, Key Laboratory of Biomedical Shanxi Province, Shanxi University, Taiyuan 030006, People's Republic of China
Cilia are organelles for cellular signalling and motility. Mutations affecting ciliary function are also associated with cilia-related disorders (ciliopathies). The identification of cilia markers is critical for studying their function at the cellular level. Due to the lack of a conserved, short ciliary localization motif, the full-length ARL13b or 5HT6 proteins are normally used for cilia labelling. Overexpression of these genes, however, can affect the function of cilia, leading to artefacts in cilia studies. Here, we show that Nephrocystin-3 (Nphp3) is highly conserved among vertebrates and demonstrate that the N-terminal truncated peptide of zebrafish Nphp3 can be used as a gratuitous cilia-specific marker. To visualize the dynamics of cilia in vivo, we generated a stable transgenic zebrafish Tg (β-actin: nphp3N-mCherry)sx1001. The cilia in multiple cell types are efficiently labelled by the encoded fusion protein from embryonic stages to adulthood, without any developmental and physiological defects. We show that the line allows live imaging of ciliary dynamics and trafficking of cilia proteins, such as Kif7 and Smo, key regulators of the Hedgehog signalling pathway. Thus, we have generated an effective new tool for in vivo cilia studies that will help shed further light on the roles of these important organelles.
