title: Complex cytogenetic analysis of early lethality mouse embryos.
abstract: An increasing interest in the molecular mechanisms governing cell division has resulted in the discovery of several groups of genes that participate in the regulation of mitosis and meiosis in eukaryotes. Inactivation of these genes in mice often leads to early embryonic lethality. To show direct causality between mutations of these genes, chromosomal instability and embryonic death, a technique enabling detailed cytogenetic analysis of embryonic cells is required. Here, we develop and test a comprehensive approach that allows complex analysis of individual early postimplantation embryos and combines polymerase chain reaction genotyping with the preparation and detailed karyotypic inspection of cells at the metaphase and anaphase stages. The method enables good chromosomal spreading and scattering of nuclei to perform routine cytogenetics (i.e., standard stain and G-banding). It also permits the application of specialized techniques such as fluorescence in situ hybridization to detect particular chromosomes and to verify the integrity of individual chromosomes. Utility of the new method is demonstrated by an analysis of embryonic day E7.5-E9.5 tissue from mice deficient in the spindle checkpoint gene Bub1b.
authors.affiliation: Transgenic Mouse model Shared Resource, Knight Cancer Institute, Oregon Health & Science University, Portland, OR 97239, USA.
authors.forename: Qi
authors.initials: Q
authors.lastname: Tian
authors.forename: Amy E
authors.initials: AE
authors.lastname: Hanlon Newell
authors.forename: Yingming
authors.initials: Y
authors.lastname: Wang
authors.forename: Susan B
authors.initials: SB
authors.lastname: Olson
authors.forename: Lev M
authors.initials: LM
authors.lastname: Fedorov
mesh_terms: D000818:Animals; D018797:Cell Cycle Proteins; D002871:Chromosome Banding; D033481:Chromosomes, Mammalian / Q000235:genetics; D020732:Cytogenetic Analysis; D020964:Embryo Loss / Q000235:genetics*; D005260:Female; D007621:Karyotyping; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D011247:Pregnancy; D017346:Protein Serine-Threonine Kinases / Q000235:genetics
chemical_list: C584278:Bub1b protein, mouse; D018797:Cell Cycle Proteins; C090464:Bub1 spindle checkpoint protein; D017346:Protein Serine-Threonine Kinases