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https://ir.csmu.edu.tw:8080/ir/handle/310902500/11642
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| Title: | The connexin 30.3 of zebrafish homologue of human connexin 26 may play similar role in the inner ear. |
| Authors: | Chang-Chien, J
YC, Yen
KH, Chien
SY, Li
TC, Hsu
Yang, JJ |
| Contributors: | 中山醫學大學 |
| Date: | 2014 |
| Issue Date: | 2015-07-27T08:04:32Z (UTC)
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| ISSN: | 0378-5955 |
| Abstract: | The intercellular gap junction channels formed by connexins (CXs) are important for recycling potassium ions in the inner ear. CXs are encoded by a family of the CX gene, such as GJB2, and the mechanism leading to mutant connexin-associated diseases, including hearing loss, remains to be elucidated. In this study, using bioinformatics, we found that two zebrafish cx genes, cx27.5 and cx30.3, are likely homologous to human and mouse GJB2. During embryogenesis, zebrafish cx27.5 was rarely expressed at 1.5-3 h post-fertilization (hpf), but a relatively high level of cx27.5 expression was detected from 6 to 96 hpf. However, zebrafish cx30.3 transcripts were hardly detected until 9 hpf. The temporal experiment was conducted in whole larvae. Both cx27.5 and cx30.3 transcripts were revealed significantly in the inner ear by reverse transcription polymerase chain reaction (RT-PCR) and whole-mount in situ hybridization (WISH). In the HeLa cell model, we found that zebrafish Cx27.5 was distributed intracellularly in the cytoplasm, whereas Cx30.3 was localized in the plasma membrane of HeLa cells stably expressing Cx proteins. The expression pattern of zebrafish Cx30.3 in HeLa cells was more similar to that of cells expressing human CX26 than Cx27.5. In addition, we found that Cx30.3 was localized in the cell membrane of hair cells within the inner ear by immunohistochemistry (IHC), suggesting that zebrafish cx30.3 might play an essential role in the development of the inner ear, in the same manner as human GJB2. We then performed morpholino knockdown studies in zebrafish embryos to elucidate the physiological functions of Cx30.3. The zebrafish cx30.3 morphants exhibited wild-type-like and heart edema phenotypes with smaller inner ears at 72 hpf. Based on these results, we suggest that the zebrafish Cx30.3 and mammalian CX26 may play alike roles in the inner ear. Thus, zebrafish can potentially serve as a model for studying hearing loss disorders that result from human CX26 mutations.
Copyright © 2014 Elsevier B.V. All rights reserved. |
| URI: | https://ir.csmu.edu.tw:8080/ir/handle/310902500/11642
http://dx.doi.org/10.1016/j.heares.2014.04.010 |
| Relation: | Hear Res. 2014 Jul;313:55-66. |
| Appears in Collections: | [免疫學研究所] 期刊論文
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