Generation of a transgenic medaka (Oryzias latipes) strain for visualization of nuclear dynamics in early developmental stages. (19th October 2016)

• Record Type: Journal Article
• Authors: Inoue, Takanobu; Iida, Atsuo; Maegawa, Shingo; Sehara‐Fujisawa, Atsuko; Kinoshita, Masato
• Is Part Of: Development growth and differentiation. Volume 58:Number 9(2016)
• Page Start: 679
• Page End: 687
• Publication Date: 2016-10-19
• Subjects: medaka -- nuclear localization signal -- pronuclear fusion; Embryology -- Periodicals; Developmental biology -- Periodicals; Growth -- Periodicals; 574.3
• DOI: 10.1111/dgd.12324
• Languages: English
• ISSNs: 0012-1592

Abstract

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. Abstract : This is the first study to label pronuclei in the living fish embryo. We used artificially designated nuclear localization signal (aNLS) fused with GFP protein as a reporter. This aNLS brings the reporter protein into pronuclear before the fusion of male and female pronuclears in living embryo.