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Showing papers by "Richard L. Gardner published in 2002"


Journal ArticleDOI
TL;DR: Findings suggest that the ectoplacental cavity may participate in maintaining proliferation throughout the developing chorionic ectoderm and, thus, in supporting its stem cell potential, and the possibility that fluid-filled cavities may play a general role in the development of tissues that border them.
Abstract: At the blastocyst stage of pre-implantation mouse development, close contact of polar trophectoderm with the inner cell mass (ICM) promotes proliferation of undifferentiated diploid trophoblast. However, ICM/polar trophectoderm intimacy is not maintained during post-implantation development, raising the question of how growth of undifferentiated trophoblast is controlled during this time. The search for the cellular basis of trophoblast proliferation in post-implantation development was addressed with an in vitro spatial and temporal analysis of fibroblast growth factor 4-dependent trophoblast stem cell potential. Two post-implantation derivatives of the polar trophectoderm - early-streak extra-embryonic ectoderm and late-streak chorionic ectoderm - were microdissected into fractions along their proximodistal axis and thoroughly dissociated for trophoblast stem cell culture. Results indicated that cells with trophoblast stem cell potential were distributed throughout the extra-embryonic/chorionic ectoderm, an observation that is probably attributable to non-coherent growth patterns exhibited by single extra-embryonic ectoderm cells at the onset of gastrulation. Furthermore, the frequency of cells with trophoblast stem cell potential increased steadily in extra-embryonic/chorionic ectoderm until the first somite pairs formed, decreasing thereafter in a manner independent of proximity to the allantois. Coincident with occlusion of the ectoplacental cavity via union between chorionic ectoderm and the ectoplacental cone, a decline in the frequency of mitotic chorionic ectoderm cells in vivo, and of trophoblast stem cell potential in vitro, was observed. These findings suggest that the ectoplacental cavity may participate in maintaining proliferation throughout the developing chorionic ectoderm and, thus, in supporting its stem cell potential. Together with previous observations, we discuss the possibility that fluid-filled cavities may play a general role in the development of tissues that border them.

132 citations


Journal ArticleDOI
TL;DR: 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.
Abstract: 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° 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.

71 citations


Journal ArticleDOI
TL;DR: The results show that the direction of polar to mural flow of cells is not random, and that the most distal mural trophectoderm cell could yield up to eight descendants during 45 h of culture.
Abstract: BACKGROUND: The present study was undertaken to ascertain whether the polarized flow of cells from polar to mural trophectoderm is related to the axis of bilateral symmetry of the blastocyst in the mouse, and whether trophectoderm cells can initiate new cycles once they have left the polar region. METHODS AND RESULTS: Two different approaches were used to investigate the relationship of polar to mural flow of trophectoderm cells to the bilateral axis. One was to mark peripheral polar trophectoderm cells at one or both ends of the bilateral axis in early blastocysts and examine the distribution of their clonal descendants after further growth in culture. The other was to mark the two ends of the bilateral axis with small oil drops in the zona pellucida in blastocysts whose polar trophectoderm was then labelled globally with fluorescent latex microspheres before culture. In both cases, marking of additional blastocysts orthogonal to the bilateral axis was also done. The results show that the direction of polar to mural flow of cells is not random, and that the most distal mural trophectoderm cell could yield up to eight descendants during 45 h of culture. CONCLUSION: The findings are consistent with the polar to mural flow of trophectoderm cells being aligned with the bilateral axis. Moreover, trophectoderm cells can embark on new cycles even when remote from the inner cell mass.

19 citations


Journal ArticleDOI
TL;DR: Regularities in axial relationships between the blastocyst and zygote have been revealed and show that, at least in normal development, patterning begins before cleavage.
Abstract: The preimplantation mammalian conceptus shows an impressive ability to develop normally following the loss, gain or rearrangement of cells. This has prompted the view that, unlike in other species, patterning in mammals cannot depend on information that is already present in the zygote before it begins to cleave. However, various findings are hard to reconcile with this conclusion, including evidence that the incidence of monozygotic twinning is sensitive to the conditions to which eggs or very early concepti are exposed. Possible causes of early twinning are discussed, and it is argued that partial hatching of the conceptus through a hernia in the zona pellucida cannot account for all cases. Moreover, it remains questionable whether studies on aggregated morulae and isolated blastomeres really provide compelling evidence against the existence of indispensible patterning information in the egg. Finally, regularities in axial relationships between the blastocyst and zygote have been revealed employing strictly non-invasive techniques. These show that, at least in normal development, patterning begins before cleavage.

8 citations