Fertile and diploid nuclear transplants derived from embryonic cells of a small laboratory fish, medaka ( Oryzias latipes )

Fertile and diploid nuclear transplants derived from embryonic cells of a small laboratory fish, medaka (Oryzias latipes)

^*Laboratory of Freshwater Fish Stocks, Bioscience Center, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan; ^‡Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan; and ^§Division of Cell Technology, National Research Institute of Aquaculture, Tamaki-cho, Watarai-gun, Mie 519-0423, Japan

Communicated by John B. Gurdon, University of Cambridge, Cambridge, United Kingdom (received for review September 21, 2000)

Abstract

Fertile and diploid nuclear transplants were successfully generated by using embryonic cells as donors in a small laboratory fish, medaka (Oryzias latipes). Embryonic cell nuclei from transgenic fish carrying the green fluorescent protein (GFP) gene were transplanted into unfertilized eggs enucleated by x-ray irradiation. In this study, 1 out of 588 eggs transplanted in the first experiment and 5 out of 298 eggs transplanted in the second experiment reached the adult stage. All of these nuclear transplants were fertile and diploid, and the natural and GFP markers of the donor nuclei were transmitted to the F₁ and F₂ offspring in a Mendelian fashion. This systematic study proves the feasibility of generating nuclear transplants by using embryonic cells from fish as donors, and it is supported by convincing evidence.

Footnotes

↵ † To whom reprint requests should be addressed. E-mail: wakamatu{at}bio.nagoya-u.ac.jp.
Abbreviations:

β-Act, β-actin gene promoter,

BSS, balanced salt solution;

EF1a-A,

elongation factor 1a-A;

GFP,

green fluorescent protein;

OR,

orange-red fish;

PGM,

phosphoglucomutase;

PS,

penicillin and streptomycin