Abstract
In mammals, the ability to successfully introduce isolated cells into a recipient embryo and to document their development has been an important experimental advance in determining the developmental potential of cells. These techniques are also useful in reestablishing the genome of embryonic cells into a germ line. The objective of the present study was to determine whether blastomeres isolated from rainbow trout (Oncorhynchus mykiss) will incorporate and continue to develop when injected into recipient embryos. In the first experiment, donor cells, previously labeled by injecting fluorescein isothiocyanate-dextran into the zygote, were isolated from blastulae; approximately 1000 of these cells were microinjected into each unlabeled recipient embryo of the same developmental stage. Following subsequent development through gastrulation, microscopic examination revealed that 19 of 114 injected embryos (17%) contained fluorescent cells. These labeled cells were present at numerous sites within embryos, and the pattern of distribution of these cells varied among embryos. In experiment two, blastomeres from normal diploid embryos were injected into triploid blastulae. The injected embryos were incubated until hatching and then sacrificed; cells from these embryos were dispersed and treated with 4',6-diamidino-2-phenylindole. The proportion of diploid cells, as determined by flow cytometry, varied from 2.0% to 12%. From these results we conclude that blastomeres isolated from rainbow trout blastulae will incorporate and continue to develop following injection into recipient embryos.
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- Anderson G. B. Use of chimaeras to study development. J Reprod Fertil Suppl. 1987;34:251–259. [PubMed] [Google Scholar]
- Bradley A., Evans M., Kaufman M. H., Robertson E. Formation of germ-line chimaeras from embryo-derived teratocarcinoma cell lines. Nature. 1984 May 17;309(5965):255–256. doi: 10.1038/309255a0. [DOI] [PubMed] [Google Scholar]
- Brinster R. L. The effect of cells transferred into the mouse blastocyst on subsequent development. J Exp Med. 1974 Oct 1;140(4):1049–1056. doi: 10.1084/jem.140.4.1049. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chourrout D. Thermal induction of diploid gynogenesis and triploidy in the eggs of the rainbow trout (Salmo gairdneri Richardson). Reprod Nutr Dev. 1980;20(3A):727–733. doi: 10.1051/rnd:19800415. [DOI] [PubMed] [Google Scholar]
- Doetschman T., Williams P., Maeda N. Establishment of hamster blastocyst-derived embryonic stem (ES) cells. Dev Biol. 1988 May;127(1):224–227. doi: 10.1016/0012-1606(88)90204-7. [DOI] [PubMed] [Google Scholar]
- Evans M. J., Kaufman M. H. Establishment in culture of pluripotential cells from mouse embryos. Nature. 1981 Jul 9;292(5819):154–156. doi: 10.1038/292154a0. [DOI] [PubMed] [Google Scholar]
- Gossler A., Doetschman T., Korn R., Serfling E., Kemler R. Transgenesis by means of blastocyst-derived embryonic stem cell lines. Proc Natl Acad Sci U S A. 1986 Dec;83(23):9065–9069. doi: 10.1073/pnas.83.23.9065. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ho R. K., Kane D. A. Cell-autonomous action of zebrafish spt-1 mutation in specific mesodermal precursors. Nature. 1990 Dec 20;348(6303):728–730. doi: 10.1038/348728a0. [DOI] [PubMed] [Google Scholar]
- Kimmel C. B., Law R. D. Cell lineage of zebrafish blastomeres. I. Cleavage pattern and cytoplasmic bridges between cells. Dev Biol. 1985 Mar;108(1):78–85. doi: 10.1016/0012-1606(85)90010-7. [DOI] [PubMed] [Google Scholar]
- Kimmel C. B., Warga R. M. Cell lineage and developmental potential of cells in the zebrafish embryo. Trends Genet. 1988 Mar;4(3):68–74. doi: 10.1016/0168-9525(88)90043-1. [DOI] [PubMed] [Google Scholar]
- Martin G. R. Isolation of a pluripotent cell line from early mouse embryos cultured in medium conditioned by teratocarcinoma stem cells. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7634–7638. doi: 10.1073/pnas.78.12.7634. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robertson E., Bradley A., Kuehn M., Evans M. Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature. 1986 Oct 2;323(6087):445–448. doi: 10.1038/323445a0. [DOI] [PubMed] [Google Scholar]
- TARKOWSKI A. K. Mouse chimaeras developed from fused eggs. Nature. 1961 Jun 3;190:857–860. doi: 10.1038/190857a0. [DOI] [PubMed] [Google Scholar]