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Richard L Gardner

Publications and source records attributed to Richard L Gardner.

9 recordsLinked to original sources
PubMed2005

The case for prepatterning in the mouse.

In studies from several laboratories using a variety of different techniques, features of the zygote and two-cell conceptus have been found to map nonrandomly on the blastocyst with respect to both its axis of polarity and bilateral plane. This is not what would be expected if, as is widely believed, early patterning depends entirely on positional relationships and interactions among the progeny of blastomeres that are equipotential until at least the eight-cell stage. Rather, the implication of these findings is that prepatterning is a normal facet of development in mammals, just as it is in most other metazoa. Nevertheless, there is still no general consensus regarding the extent to which such prepatterning depends on intrinsic organization of the oocyte, as opposed to events that are contingent on fertilization.

Animals↗
PubMed2005

Experimental analysis of the transdifferentiation of visceral to parietal endoderm in the mouse.

The visceral endoderm (VE) of isolated extraembryonic regions (ExEmbs) of 7 days postcoitum (dpc) prestreak mouse conceptuses have been shown to convert readily to parietal endoderm (PE). The present study addresses the following three unanswered questions. On what does conversion depend, how rapidly does it occur, and is it an enduring general property of a residual small population of relatively immature cells? In situ hybridization reveals that change in cell state occurs within 2 days of culture. Deprivation of the mesoderm also promotes it in later ExEmbs. Conversely, the conversion to PE in isolated 7 dpc ExEmbs is suppressed by grafting 8 dpc or 9 dpc mesoderm. Hence, the conversion provides an example of transdifferentiation that is promoted by the absence of extraembryonic mesoderm. The presence of mesoderm seems to be necessary to enable the VE to grow rather than convert to PE, as occurs if it retains contact with the extraembryonic ectoderm.

Animals↗
PubMed2005

Imprinted X-inactivation in extra-embryonic endoderm cell lines from mouse blastocysts.

The extra-embryonic endoderm lineage plays a major role in the nutritive support of the embryo and is required for several inductive events, such as anterior patterning and blood island formation. Blastocyst-derived embryonic stem (ES) and trophoblast stem (TS) cell lines provide good models with which to study the development of the epiblast and trophoblast lineages, respectively. We describe the derivation and characterization of cell lines that are representative of the third lineage of the blastocyst -extra-embryonic endoderm. Extra-embryonic endoderm (XEN) cell lines can be reproducibly derived from mouse blastocysts and passaged without any evidence of senescence. XEN cells express markers typical of extra-embryonic endoderm derivatives, but not those of the epiblast or trophoblast. Chimeras generated by injection of XEN cells into blastocysts showed exclusive contribution to extra-embryonic endoderm cell types. We used female XEN cells to investigate the mechanism of X chromosome inactivation in this lineage. We observed paternally imprinted X-inactivation, consistent with observations in vivo. Based on gene expression analysis, chimera studies and imprinted X-inactivation, XEN cell lines are representative of extra-embryonic endoderm and provide a new cell culture model of an early mammalian lineage.

Animals↗
PubMed2003

The basis and significance of pre-patterning in mammals.

The second polar body (Pb) provides an enduring marker of the animal pole of the zygote, thereby revealing that the axis of bilateral symmetry of the early blastocyst is aligned with the zygote's animal-vegetal axis. That this relationship is biologically significant appeared likely when subsequent studies showed that the equator of the blastocyst tended to correspond with the plane of first cleavage. However, this cleavage plane varies both with respect to the position of the second Pb and to the distribution of components of the fertilizing sperm that continue to mark the point where it entered the egg. It also maps too variably on the blastocyst to play a causal role in early patterning. The zygote has been found transiently to exhibit bilateral symmetry before regaining an essentially spherical shape prior to first cleavage. Marking experiments indicate that the plane of bilateral symmetry of the blastocyst is aligned with, and the plane of first cleavage is typically orthogonal to, the zygote's bilateral plane. The bilateral symmetry of the zygote bears no consistent relationship either to the point of sperm entry or to the distribution of the pronuclei, and may therefore be a manifestation of intrinsic organization of the egg. Finally, the two-cell blastomere inheriting the sperm entry point has not been found to differ consistently in fate from the one that does not.

Animals↗
PubMed2003

The derivation of highly germline-competent embryonic stem cells containing NOD-derived genome.

It would be extremely advantageous to the analysis of disease mechanisms in the spontaneous mouse model of type 1 diabetes, the nonobese diabetic (NOD) strain, if genes in this strain could be modified in vivo using embryonic stem (ES) cells and homologous recombination. However, a NOD ES cell line with adequate germline transmission has not yet been reported. We report the development of highly germline-competent ES cell lines from the F1 hybrid of NOD and 129 for use in NOD gene targeting. Consequently, we developed ES cell lines derived from (NOD x 129)F1 x 129 backcross 1 mice, which were intercrossed to select for homozygosity of particular regions of NOD genome known to contain disease loci.

Animals↗
PubMed2002

Trophectoderm growth and bilateral symmetry of the blastocyst in the mouse.

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.

Animals↗
PubMed2002

Inhibition of trophoblast stem cell potential in chorionic ectoderm coincides with occlusion of the ectoplacental cavity in the mouse.

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.

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