Visualizing the translational activation of a particular mRNA in zebrafish embryos using in situ hybridization and proximity ligation assay.
| Autor: | Sato K; Biosystems Science Course, Graduate School of Life Science, Hokkaido University, Sapporo 060-0810, Japan. Electronic address: sato.keisuke.b7@elms.hokudai.ac.jp., Kotani T; Biosystems Science Course, Graduate School of Life Science, Hokkaido University, Sapporo 060-0810, Japan; Department of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo 060-0810, Japan. Electronic address: tkotani@sci.hokudai.ac.jp. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2024 Jun 21; Vol. 5 (2), pp. 102951. Date of Electronic Publication: 2024 Mar 15. |
| DOI: | 10.1016/j.xpro.2024.102951 |
| Abstrakt: | Fertilized eggs initiate translation of stored mRNAs in spatially and temporally controlled manners. Here, we present a protocol for visualizing spatial and temporal translation in zebrafish embryos by fluorescence in situ hybridization and proximity ligation assay. We describe steps for labeling newly synthesized proteins and mRNA, visualizing mRNA translation and mRNA, sample mounting, and observation. Coupling detection of mRNA molecules with their translation sites is useful for understanding the molecular and cellular mechanisms that drive embryo development. For complete details on the use and execution of this protocol, please refer to Sato et al. 1 and Takada et al. 2 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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