title: Three-dimensional reconstruction of tetraploid<-->diploid chimaeric mouse blastocysts.
abstract: Studies of tetraploid<-->diploid (4n<-->2n) mouse chimaeras have demonstrated unequal contributions of 4n cells to different tissues of the midgestation conceptus. Such a pattern has also been reported in chimaeras as early as E3.5d, which show an enhanced contribution of 4n cells to the mural trophectoderm (Everett & West, 1996). In this study, sectioned 4n<-->2n and 2n<-->2n control chimaeric blastocysts were digitised and reconstructed in 3 dimensions (3-D). The 3-D images revealed only limited mixing of cells from the 2 contributing embryos of individual blastocysts in both chimaera groups. Consequently, the distribution pattern of the 2 cell types was dependent on the spatial relationship between the orientation of the blastocyst and the boundary between the 2 clusters of cells. The distribution patterns observed were not strikingly different for 4n<-->2n and 2n<-->2n chimaeras, each showing some transgenic positive cell contribution in all 3 identifiable developmental lineages. It was notable, however, that in all 4n<-->2n blastocysts at least some 4n cells were located adjacent to the blastocyst cavity. Such a consistent pattern was not evident in 2n<-->2n chimaeras. This study has demonstrated the value of 3-D reconstructions for the analysis of spatial relationships of 2 cell populations in chimaeric mouse blastocysts.
authors.affiliation: Department of Reproductive and Developmental Sciences, University of Edinburgh, Western General Hospital, UK.
authors.forename: C A
authors.initials: CA
authors.lastname: Everett
authors.forename: M H
authors.initials: MH
authors.lastname: Stark
authors.forename: J D
authors.initials: JD
authors.lastname: West
authors.forename: D
authors.initials: D
authors.lastname: Davidson
authors.forename: R A
authors.initials: RA
authors.lastname: Baldock
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000648:ultrastructure*; D019070:Cell Lineage; D002678:Chimera; D004171:Diploidy; D005865:Gestational Age; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008854:Microscopy, Electron; D011003:Ploidies; D011123:Polyploidy