title: Sexing whole human pre-embryos by in-situ hybridization with a Y-chromosome specific DNA probe.
abstract: We have used DNA-DNA in-situ hybridization with a DNA probe for the human Y-chromosome to distinguish between male and female human pre-embryos. Both biotinylated and tritiated Y-probes worked well on control cell cultures where 100 interphase nuclei were scored. Morphologically normal pre-embryos could be sexed with confidence with the tritiated Y-probe but the biotin results were less reliable (although only a few pre-embryos were analysed in this way). Early cleavage stage pre-embryos had large nuclei with relatively diffuse Y-bodies and were more difficult to score with the biotinylated Y-probe. Morphologically abnormal pre-embryos often had large nuclei with multiple Y-bodies (presumably polyploid nuclei) or small nuclei with no Y-bodies (possibly fragmenting nuclei). In all, 38 cleaving and two non-cleaving pre-embryos were analysed. The incidence of false positive and false negative cells seen after hybridization of tritiated Y-probes to control lymphocyte cultures suggests that it should normally be possible to distinguish morphologically normal male and female pre-embryos with samples of three to six interphase nuclei.
authors.affiliation: Department of Obstetrics and Gynaecology, University of Edinburgh, UK.
authors.forename: J D
authors.initials: JD
authors.lastname: West
authors.forename: J R
authors.initials: JR
authors.lastname: Gosden
authors.forename: R R
authors.initials: RR
authors.lastname: Angell
authors.forename: K M
authors.initials: KM
authors.lastname: West
authors.forename: A F
authors.initials: AF
authors.lastname: Glasier
authors.forename: S S
authors.initials: SS
authors.lastname: Thatcher
authors.forename: D T
authors.initials: DT
authors.lastname: Baird
mesh_terms: D001755:Blastocyst / Q000032:analysis*; D015342:DNA Probes; D005260:Female; D006801:Humans; D008297:Male; D009693:Nucleic Acid Hybridization; D012732:Sex Determination Analysis; D014998:Y Chromosome / Q000032:analysis*
chemical_list: D015342:DNA Probes