Generation of a transgenic medaka (Oryzias latipes) strain for visualization of nuclear dynamics in early developmental stages |
| |
| Authors: | Shingo Maegawa Atsuko Sehara‐Fujisawa Masato Kinoshita |
| |
| Affiliation: | 1. Department of Intelligence Science and Technology, Graduate School of Informatics, Kyoto University, Sakyo‐ku, Kyoto, Japan;2. Department of Growth Regulation, Institute for Frontier Medical Sciences, Kyoto University, Sakyo‐ku, Kyoto, Japan;3. Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Sakyo‐ku, Kyoto, Japan |
| |
| Abstract: | In this study, we verified nuclear transport activity of an artificial nuclear localization signal (aNLS) in medaka fish (Oryzias latipes). We generated a transgenic medaka strain expresses the aNLS tagged enhanced green fluorescent protein (EGFP) driven by a medaka beta‐actin promoter. The aNLS‐EGFP was accumulated in the nuclei of somatic tissues and yolk nuclei of oocytes, but undetectable in the spermatozoa. The fluorescent signal was observed from immediately after fertilization by a maternal contribution. Furthermore, male and female pronuclei were visualized in fertilized eggs, and nuclear dynamics of pronuclear fusion and subsequent cleavage were captured by time‐lapse imaging. In contrast, SV40NLS exhibited no activity of nuclear transport in early embryos. In conclusion, the aNLS possesses a strong nuclear localization activity and is a useful probe for fluorescent observation of the pronuclei and nuclei in early developmental stage of medaka. |
| |
| Keywords: | medaka nuclear localization signal pronuclear fusion |
|
|
