Effect of donor cell cycle stage on chromatin and spindle morphology in nuclear transplant rabbit embryos.

We investigated the influence of the cell cycle stage of the nuclear donor on prematurely condensed chromatin (PCC) and spindle morphology and on chromosome constitution in rabbit nuclear transplant embryos. The configuration of PCC following nuclear transplantation with G1, early S, and late S phase donor nuclei (G1, early S, and late S transplants, respectively) was characterized in whole mounts and chromosome spreads. In addition, the influence of the donor cell cycle stage on chromosome constitution in cleavage stage-manipulated embryos was determined. Within 2 h after fusion of the donor blastomere, the recipient oocyte cytoplasm was able to induce formation de novo of a metaphase plate associated with a spindle in G1, early S, and late S transplants. Metaphase chromosomes and spindle were intact in most cases of PCC in G1 transplants. However, these structures displayed minor abnormalities in early S transplants and gross abnormalities in late S transplants, such as incomplete or absent spindle formation and incomplete chromatin condensation. Normal chromosomes were present in G1 and early S transplants, whereas chromosome abnormalities were detected in late S transplants. The results indicate that morphology of prematurely condensed G1 and early S chromatin has a minor influence on chromosome constitution of manipulated embryos. That of late S chromatin, however, affects chromosome constitution in embryos and may account for reduced development of nuclear transplant embryos when late S phase donor nuclei are used.

[1]  P. Collas,et al.  Influence of cell cycle stage of the donor nucleus on development of nuclear transplant rabbit embryos. , 1992, Biology of reproduction.

[2]  P. Collas,et al.  Relationship between nuclear remodeling and development in nuclear transplant rabbit embryos. , 1991, Biology of reproduction.

[3]  P. Collas,et al.  Factors affecting the efficiency of nuclear transplantation in the rabbit embryo. , 1990, Biology of reproduction.

[4]  G Hermanson,et al.  High-resolution mapping of human chromosome 11 by in situ hybridization with cosmid clones. , 1990, Science.

[5]  A. Tarkowski,et al.  Remodeling of mouse thymocyte nuclei depends on the time of their transfer into activated, homologous oocytes. , 1988, Journal of cell science.

[6]  S. Stice,et al.  Nuclear reprogramming in nuclear transplant rabbit embryos. , 1988, Biology of reproduction.

[7]  Y. Matsuda,et al.  Chromosomal analysis in mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) and methyl and ethyl methanesulfonate (MMS and EMS). , 1988, Mutation research.

[8]  A. Tarkowski,et al.  Remodelling of thymocyte nuclei in activated mouse oocytes: an ultrastructural study. , 1986, European journal of cell biology.

[9]  A. Tarkowski,et al.  Chromosome condensation activity in ovulated metaphase II mouse oocytes assayed by fusion with interphase blastomeres. , 1986, Journal of cell science.

[10]  G. Schatten,et al.  Microtubule configurations during fertilization, mitosis, and early development in the mouse and the requirement for egg microtubule-mediated motility during mammalian fertilization. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[11]  A. Tarkowski,et al.  Behaviour of thymocyte nuclei in non-activated and activated mouse oocytes. , 1984, Journal of cell science.

[12]  R. Brown,et al.  Genetic manipulation by means of microcell-mediated transfer of normal human chromosomes into recipient mouse cells. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[13]  R. Johnson,et al.  The induction of DNA synthesis in the chick red cell nucleus in heterokaryons during the first cell cycle after fusion with HeLa cells. , 1975, Journal of cell science.

[14]  H. Klinger,et al.  Modulation of the activity of an avian gene transferred into a mammalian cell by cell fusion. , 1974, Proceedings of the National Academy of Sciences of the United States of America.

[15]  H. Harris,et al.  Correction of genetic defects in mammalian cells by the input of small amounts of foreign genetic material. , 1973, Journal of cell science.

[16]  H. Harris,et al.  Correction of a genetic defect in a mammalian cell. , 1971, Nature: New biology.

[17]  Robert T. Johnson,et al.  Mammalian cell fusion III. A hela cell inducer of premature chromosome condensation active in cells from a variety of animal species , 1970 .

[18]  P. N. Rao,et al.  Mammalian Cell Fusion : Induction of Premature Chromosome Condensation in Interphase Nuclei , 1970, Nature.

[19]  Robert T. Johnson,et al.  Mammalian Cell Fusion : Studies on the Regulation of DNA Synthesis and Mitosis , 1970, Nature.

[20]  Y. Masui,et al.  Chromosome condensation activity in the cytoplasm of anucleate and nucleate fragments of mouse oocytes. , 1986, Developmental biology.