A transgenic zebrafish for in vivo visualization of cilia
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 ci...
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| Format: | Article |
| Language: | English |
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The Royal Society
2022-08-01
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| Series: | Open Biology |
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| Online Access: | https://royalsocietypublishing.org/doi/10.1098/rsob.220104 |
| _version_ | 1797840267566383104 |
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| author | Hongyu Zhang Zhuoya Huang Liuliu LV Yuye Xin Qian Wang Feng Li Lina Dong Changxin Wu Philip W. Ingham Zhonghua Zhao |
| author_facet | Hongyu Zhang Zhuoya Huang Liuliu LV Yuye Xin Qian Wang Feng Li Lina Dong Changxin Wu Philip W. Ingham Zhonghua Zhao |
| author_sort | Hongyu Zhang |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-09T16:12:09Z |
| format | Article |
| id | doaj.art-4d9cc8d1af2649909f77cefea14d0f8f |
| institution | Directory Open Access Journal |
| issn | 2046-2441 |
| language | English |
| last_indexed | 2024-04-09T16:12:09Z |
| publishDate | 2022-08-01 |
| publisher | The Royal Society |
| record_format | Article |
| series | Open Biology |
| spelling | doaj.art-4d9cc8d1af2649909f77cefea14d0f8f2023-04-24T09:16:43ZengThe Royal SocietyOpen Biology2046-24412022-08-0112810.1098/rsob.220104A transgenic zebrafish for in vivo visualization of ciliaHongyu Zhang0Zhuoya Huang1Liuliu LV2Yuye Xin3Qian Wang4Feng Li5Lina Dong6Changxin Wu7Philip W. Ingham8Zhonghua Zhao9Institute 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 ChinaInstitute 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 ChinaInstitute 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 ChinaInstitute 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 ChinaInstitute 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 ChinaDepartment of Molecular Biology, Shanxi Cancer Hospital, Affiliated Cancer Hospital of Shanxi Medical University, Taiyuan 030013, People's Republic of ChinaCentral Laboratory, Shanxi Provincial People's Hospital, Affiliate of Shanxi Medical University, Taiyuan 030012, People's Republic of ChinaInstitute 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 ChinaLKC Medicine School, Nanyang Technological University, 639798, SingaporeInstitute 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 ChinaCilia 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.https://royalsocietypublishing.org/doi/10.1098/rsob.220104transgenic zebrafishciliaNphp3live imagingciliary signal peptides |
| spellingShingle | Hongyu Zhang Zhuoya Huang Liuliu LV Yuye Xin Qian Wang Feng Li Lina Dong Changxin Wu Philip W. Ingham Zhonghua Zhao A transgenic zebrafish for in vivo visualization of cilia Open Biology transgenic zebrafish cilia Nphp3 live imaging ciliary signal peptides |
| title | A transgenic zebrafish for in vivo visualization of cilia |
| title_full | A transgenic zebrafish for in vivo visualization of cilia |
| title_fullStr | A transgenic zebrafish for in vivo visualization of cilia |
| title_full_unstemmed | A transgenic zebrafish for in vivo visualization of cilia |
| title_short | A transgenic zebrafish for in vivo visualization of cilia |
| title_sort | transgenic zebrafish for in vivo visualization of cilia |
| topic | transgenic zebrafish cilia Nphp3 live imaging ciliary signal peptides |
| url | https://royalsocietypublishing.org/doi/10.1098/rsob.220104 |
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