Experimental analysis of second cleavage in the mouse.

BACKGROUND Mammalian conceptuses typically have an approximately regular tetrahedral shape at the 4-cell stage. In the rabbit, this has been attributed to both 2-cell blastomeres dividing meridionally, but with the animal-vegetal axis of the second blastomere to divide rotating through roughly 90 degrees before or during cytokinesis. The aim of the present study was to ascertain whether this was also true for the mouse. METHODS AND RESULTS First, the distribution in regular tetrahedral 4-cell conceptuses of fluorescent microspheres applied to the vegetal polar region of one or both blastomeres at the 2-cell stage was analysed. Second, the ability of 2-cell stages to form regular tetrahedral 4-cell conceptuses after the previtelline space had been gelated to prevent blastomeres from rotating was also investigated. Neither experiment yielded evidence supporting blastomere rotation during second cleavage. Rather, the findings were consistent with the regular tetrahedral form of 4-cell conceptus resulting from meridional division of one blastomere and approximately equatorial division of the other. CONCLUSIONS Second cleavage in the mouse typically yields 4-cell conceptuses with three distinct types of blastomere. While both products of the meridional division include all axial levels of the zygote, those of the equatorial division acquire only its vegetal or animal half.

[1]  M. Zernicka-Goetz,et al.  Sperm entry position provides a surface marker for the first cleavage plane of the mouse zygote , 2002, Genesis.

[2]  D. L. Dorr,et al.  Determining the stage of the estrous cycle in the mouse by the appearance of the vagina. , 1973, Biology of reproduction.

[3]  R. Gardner,et al.  Trophectoderm growth and bilateral symmetry of the blastocyst in the mouse. , 2002, Human reproduction.

[4]  R. Gardner,et al.  The early blastocyst is bilaterally symmetrical and its axis of symmetry is aligned with the animal-vegetal axis of the zygote in the mouse. , 1997, Development.

[5]  E. Borghese,et al.  Cleavage of Mouse Egg , 1963 .

[6]  H. Suzuki,et al.  Developmental ability of zona-free mouse embryos is influenced by cell association at the 4-cell stage. , 1995, Biology of reproduction.

[7]  A. Dalcq Introduction to general embryology , 1957 .

[8]  A. Tarkowski,et al.  Development of blastomeres of mouse eggs isolated at the 4- and 8-cell stage. , 1967, Journal of embryology and experimental morphology.

[9]  Z. A. Deussen,et al.  Features of cell lineage in preimplantation mouse development. , 1978, Journal of embryology and experimental morphology.

[10]  M. Johnson,et al.  The nature of intercellular coupling within the preimplantation mouse embryo. , 1984, Journal of embryology and experimental morphology.

[11]  Y. Tsunoda,et al.  Effect of cutting the zona pellucida on the pronuclear transplantation in the mouse. , 1986, The Journal of experimental zoology.

[12]  R. Gardner,et al.  Can developmentally significant spatial patterning of the egg be discounted in mammals? , 1996, Human reproduction update.

[13]  R. Pedersen,et al.  Polarity of the mouse embryo is anticipated before implantation. , 1999, Development.

[14]  T. Davies,et al.  The plane of first cleavage is not related to the distribution of sperm components in the mouse. , 2002, Human reproduction.

[15]  R. Gardner 2 – Asymmetry and Prepattern in Mammalian Development , 2002 .

[16]  R. Gardner,et al.  Specification of embryonic axes begins before cleavage in normal mouse development. , 2001, Development.

[17]  J. Rossant,et al.  Postimplantation development of blastomeres isolated from 4- and 8-cell mouse eggs. , 1976, Journal of embryology and experimental morphology.

[18]  M. Depew,et al.  Mouse Development: Patterning, Morphogenesis, and Organogenesis , 2002 .

[19]  R. Pedersen,et al.  Blastomeres arising from the first cleavage division have distinguishable fates in normal mouse development. , 2001, Development.

[20]  A. Trounson,et al.  Allocation of cells to the inner cell mass and trophectoderm of 3/4 mouse embryos. , 1990, Reproduction, fertility, and development.

[21]  J. Biggers,et al.  Fertilization in vitro of mouse ova from inbred and outbred strains: complete preimplantation embryo development in glucose-supplemented KSOM. , 1995, Biology of reproduction.

[22]  R. Gardner Axial relationships between egg and embryo in the mouse. , 1998, Current topics in developmental biology.

[23]  L. J. Smith,et al.  Embryonic axis orientation in the mouse and its correlation with blastocyst relationships to the uterus. Part 1. Relationships between 82 hours and 4 1/4 days. , 1980, Journal of embryology and experimental morphology.

[24]  G. Rose Cinemicrography in cell biology , 1963 .

[25]  J. Biggers,et al.  IVF of mouse ova in a simplex optimized medium supplemented with amino acids. , 2000, Human reproduction.

[26]  Magdalena Zernicka-Goetz,et al.  Role for sperm in spatial patterning of the early mouse embryo , 2001, Nature.

[27]  J. Mulnard,et al.  Cell division and cell allocation in early mouse development. , 1978, Journal of embryology and experimental morphology.

[28]  A. Solomko,et al.  Role of ooplasmic segregation in mammalian development , 2004, Roux's archives of developmental biology.

[29]  R. Gardner,et al.  Effect of damage to the zona pellucida on development of preimplantation embryos in the mouse. , 1989, Human reproduction.

[30]  R. Gardner,et al.  Mouse chimeras and the analysis of development. , 2000, Methods in molecular biology.

[31]  B. Gulyas A reexamination of cleavage patterns in eutherian mammalian eggs: rotation of blastomere pairs during second cleavage in the rabbit. , 1975, The Journal of experimental zoology.

[32]  S. Howlett,et al.  Sequence and regulation of morphological and molecular events during the first cell cycle of mouse embryogenesis. , 1985, Journal of embryology and experimental morphology.

[33]  D. Altman,et al.  Is the anterior-posterior axis of the fetus specified before implantation in the mouse? , 1992, The Journal of experimental zoology.