scholarly journals Mouse zygotes as recipients in embryo cloning

Reproduction ◽  
2006 ◽  
Vol 132 (5) ◽  
pp. 741-748 ◽  
Author(s):  
Pawel Gręda ◽  
Jolanta Karasiewicz ◽  
Jacek A Modliński
Keyword(s):  
Nuclear Membrane ◽  
Genetic Material ◽  
Coat Colour ◽  
New Technique ◽  
Glucose Phosphate Isomerase ◽  
Cell Stage ◽  
Glucose Phosphate ◽  
Mouse Zygotes ◽  
A New Technique ◽  
Embryo Cloning

Zygotes have not been recognized as nuclear recipients since enucleated zygotes receiving nuclei from beyond two-cell stage embryos are not able to form blastocysts. In the present study, a new technique of zygote enucleation is presented, which consists in selectively removing the nuclear membrane with genetic material of pronuclei, but leaving other pronuclear components in the cytoplasm. With selective enucleation it is possible – after transfer of eight-cell stage nuclei – to obtain 70.5 and 7.8% of preimplantation and full-term development respectively. Origin of cloned mice from introduced nuclei was confirmed by the coat colour and glucose phosphate isomerase (GPI) isozyme of the donor. We suggest that some pronuclear factors – taken away from the zygotes in the karyoplasts upon classical enucleation – are needed to reprogram the introduced nuclei.

Germ cell nuclei of male fetal mice can support development of chimeras to midgestation following serial transplantation

Development ◽  
1995 ◽  
Vol 121 (3) ◽  
pp. 779-783 ◽  
Author(s):  
Y. Kato ◽  
Y. Tsunoda
Keyword(s):  
Germ Cells ◽  
Nuclear Transfer ◽  
Yolk Sac ◽  
Glucose Phosphate Isomerase ◽  
Cell Nuclei ◽  
Cell Stage ◽  
Glucose Phosphate ◽  
Fetal Mice ◽  
Serial Nuclear Transfer

Chimeric embryos between fertilized eggs from F1 (C57BL × CBA) and 15.5-16.5 days post coitum (dpc) male fetal germ cells (FGCs) from CD-1 strain (glucose phosphate isomerase, Gpi-1a/a) mice were produced by nuclear transfer. Briefly, a single FGC was fused with enucleated oocytes and activated, and the reconstituted oocytes were cultured to the 2-cell stage. The nucleus from the reconstituted 2-cell embryos was then transferred into an enucleated blastomere of the same stage embryos derived from F1 mice to produce chimeric embryos. The reconstituted 2-cell embryos, which synchronously divided to the 4-cell stage after treatment with nocodazole, were further cultured in vitro. Compacted morula and blastocysts were transferred to the uteri of pseudopregnant female mice. Some recipients were allowed to develop to term and the others were killed at mid gestation to analyze the contribution of donor FGC-derived cells. Survival to term was low with no chimeric animals. Glucose phosphate isomerase (GPI) analysis at midgestation revealed that some conceptuses had chimerism in the fetuses, trophoblast and yolk sac at day 10.5 of pregnancy. The contribution of donor cells was 37–47%, 19–65% and 12–63%, respectively. It was concluded that the nucleus from 15.5-16.5 dpc male fetal germ cells had the potency to develop into fetus, trophoblast and yolk sac after serial nuclear transfer with oocytes and fertilized embryos. The reason for the low viability of chimeric embryos is discussed.

A new technique for the mapping of oxygen tension on the brain surface

2005 ◽  
Vol 25 (1_suppl) ◽  
pp. S543-S543
Author(s):  
Satoshi Kimura ◽  
Keigo Matsumoto ◽  
Yoshio Imahori ◽  
Katsuyoshi Mineura ◽  
Toshiyuki Itoh
Keyword(s):  
Oxygen Tension ◽  
New Technique ◽  
Brain Surface ◽  
A New Technique ◽  
The Brain

A new technique to visualize alveolar dynamics in a rabbit model

2009 ◽  
Vol 56 (S 01) ◽  
Author(s):  
J Bickenbach ◽  
R Rossaint ◽  
R Autschbach ◽  
R Dembinski
Keyword(s):  
Rabbit Model ◽  
New Technique ◽  
A New Technique
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