Synchronous division of mouse two-cell embryos with nocodazole in vitro

Kato, Yuki; Tsunoda, Yukio

doi:10.1530/jrf.0.0950039

Cited by 53 publications

(26 citation statements)

References 8 publications

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“…Exposure of embryos to the concentration of nocodazole necessary to cause cell cycle arrest does not affect the ability of the manipulated embryos to complete development to term (ref. 27 and unpublished data). Thus, large numbers of identical embryos can be produced from the donor nuclei.…”

Section: Discussionmentioning

confidence: 68%

Production of identical sextuplet mice by transferring metaphase nuclei from four-cell embryos

Kwon

Kono

1996

Proc. Natl. Acad. Sci. U.S.A.

103

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Mouse clones were produced by serial nuclear transfer commencing with the transfer of four-cell nuclei at metaphase into unfertilized ooplasts. The donor four-cellstage nuclei were synchronized in metaphase with nocodazole. The oocytes receiving a four-cell nucleus at metaphase formed two nuclei after artificial activation and inhibition of cytokinesis with cytochalasin B. To obtain embryos with diploid sets of chromosomes, nuclei from each reconstructed embryo were transferred individually into separate enucleated fertilized one-cell embryos, thus doubling the number of identical embryos. This procedure produced a high frequency of development of reconstructed embryos to the blastocyst stage. Of 11 sets of identical embryos produced by serial nuclear transplantation, 83% developed into blastocysts, including three sets of identical septuplet blastocysts. After transfer to recipient mice, a total of 25 (57%) live young were obtained, which included one set of identical sextuplet and two sets of identical quadruplet mice.

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Section: Discussionmentioning

confidence: 68%

Production of identical sextuplet mice by transferring metaphase nuclei from four-cell embryos

Kwon

Kono

1996

Proc. Natl. Acad. Sci. U.S.A.

103

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“…Mimosine is a plant amino acid that inhibits the initiation of replication and blocks cell cycle in the middle-late G 1 phase (29,30), whereas OH-urea inhibits DNA replication by interfering with DNA polymerases (31) and stops cell cycle at the beginning of the S phase after crossing the restriction point (32). Nocodazole inhibits the cell cycle of macrophages at the G 2 -M phase by blocking the polimerization of tubuline needed for the microspindle formation (33,34).…”

Section: Resultsmentioning

confidence: 99%

The Expression of MHC Class II Genes in Macrophages Is Cell Cycle Dependent

Xaus

Comalada

Barrachina

et al. 2000

The Journal of Immunology

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Using different drugs, we stopped the cell cycle of bone marrow-derived macrophages at different points. After IFN-γ stimulation, macrophages arrested at the G1 phase of the cell cycle did not increase cell surface expression of the MHC class II IA. This inhibition is specific, because, under the same conditions, IFN-γ induces the expression of Fcγ receptors and the inducible NO synthase mRNA. Treatments that inhibit macrophage proliferation by blocking the cell cycle at the G1 phase, such as adenosine, forskolin, or LPS, blocked the IFN-γ induction of IA. Under IFN-γ treatment, the steady-state levels of IAα and IAβ mRNA did not increase in cells arrested at the G1 phase and the half-life of the MHC mRNA was not modified. These data suggest that the cell cycle modulation of IFN-γ-induced MHC II gene expression occurs at the transcriptional level. The expression of the class II transactivator mRNA induced by IFN-γ was also blocked when macrophages were arrested at the G1 phase of the cell cycle, suggesting that the lack of IFN-γ response occurs at the early steps of MHC class II expression. Finally, macrophages arrested at the G1 phase showed increased basal levels of cell surface IA due to an increase of the translational efficiency. These data show that the expression of MHC class II genes is regulated by the cell cycle.

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“…Mouse preimplantation embryos are easily synchronized at metaphase of the cell cycle by culturing the embryos in a medium with nocodazole. The exposure of embryos to the concentration of nocodazole necessary to cause cell cycle arrest does not affect the ability of the manipulated embryos to complete development to term [30,31]. Therefore, synchronization of donor nuclei at M phase of the cell cycle may be suitable for improving development of nuclear transplants.…”

Section: Discussionmentioning

confidence: 99%

Production of Live Young by Serial Nuclear Transfer with Mitotic Stages of Donor Nuclei in Mice.

Kwon

Kono

Nakahara

1997

Journal of Reproduction and Development

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Abstract. The developmental ability of reconstituted embryos that received nuclei from 2-, 4-and 8-cell embryos at M phase of the cell cycle was examined in vitro and in vivo. After fusion of M phase nuclei with enucleated oocytes, the reconstituted oocytes were artificially activated and cultured with cytochalasin B to achieve formation of two "pronuclei-like nuclei" (PN-like nuclei). To obtain embryos with a diploid set of chromosomes, nuclei from each reconstructed embryo were transferred individually into separate enucleated fertilized 1-cell embryos. These nuclear transplants developed to multicellular stages at higher rates and the number of reconstituted embryos doubled. Of the embryos reconstituted with 2-, 4-, and 8-cell nuclei at M phase, 26 (23/88), 40 (32/ 80) and 4% (5/135) developed to blastocysts, respectively. Live young were produced from blastocysts obtained from reconstituted embryos that received 2-and 4-cell nuclei at M phase, after transfer into recipient females. to be a result of exchanges of nuclear proteins between the nuclear and cytoplasmic compartments [12]. Several nuclear transfer systems have been proposed that produce reconstituted embryos from donor nuclei containing a normal diploid set of chromosomes at different stages of the cell cycle [13,14]. The key is the relational timing between fusion of the donor nucleus with an enucleated oocyte and activation of the oocyte [14,15]. The reconstituted oocytes extrude an extra-polar body after receiving stimuli that induce oocyte activation. To develop, reconstituted oocytes must contain a normal set of diploid chromosomes after fusion with a donor nucleus and activation. We developed an efficient procedure for transplanting G2 phase nuclei into enucleated mouse oocytes. After activation, the oocytes emitted an extra-polar body and formed a normal diploid nucleus. It was revealed that G2 phase nuclei from 2-cell stage C enucleated oocytes, e.g., mice [1,2]; rabbits [3]; cattle [4,5]; pigs [6]; sheep [7]. The transplantation of an interphase nucleus into enucleated metaphase II oocytes leads to the remodeling of the donor nucleus; nuclear envelope breakdown (NEBD) and premature chromosome condensation (PCC). Interphase nuclei transferred into oocytes undergo PCC, which is associated with the spindle apparatus that is induced by the high level of maturation promoting factor (MPF) activity in the oocyte. When reconstituted oocytes receive suitable stimuli for parthenogenetic activation, extrusion of an extra-polar body [1,7], chromatin decondensation and nuclear reformation occur [8][9][10][11]. Nuclear remodeling of the reconstituted oocytes is thought

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Synchronous division of mouse two-cell embryos with nocodazole in vitro

Cited by 53 publications

References 8 publications

Production of identical sextuplet mice by transferring metaphase nuclei from four-cell embryos

Production of identical sextuplet mice by transferring metaphase nuclei from four-cell embryos

The Expression of MHC Class II Genes in Macrophages Is Cell Cycle Dependent

Production of Live Young by Serial Nuclear Transfer with Mitotic Stages of Donor Nuclei in Mice.

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