Molecular Human Reproduction, Vol. 9, No. 10, 577-585,
October 2003
© 2003 European Society of Human Reproduction and Embryology
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Sequential multiple probe fluorescence in-situ hybridization analysis of human oocytes and polar bodies by combining centromeric labelling and whole chromosome painting
Submitted on March 14, 2003; accepted on May 20, 2003
1 Service de Génétique Moléculaire et Chromosomique, C.H.U. Arnaud de Villeneuve, 34295 Montpellier, Cedex 5, 2 CNRS UPR 1142, Institut de Génétique Humaine, 141 rue de la Cardonille, 34396 Montpellier Cedex 5 and 3 Laboratoire de Biologie de la Reproduction, Clinique St Roch, Montpellier, France
4 To whom correspondence should be addressed. e-mail: franck.pellestor{at}igh.cnrs.fr
The incidence of chromosomal aneuploidy was analysed in 104 unfertilized human oocytes and 56 first polar bodies using a double-label fluorescence in-situ hybridization (FISH) procedure. Combinations of centromeric (or locus-specific) DNA probes and whole chromosome painting probes for chromosomes 9, 13, 16, 18, 21 and X were applied on oocyte preparations, in a sequential FISH protocol. This combined approach allowed a precise in-situ identification of both chromosomes and free chromatids, and consequently a reliable analysis of chromosomal segregation errors. Of the 104 analysed oocytes, 84 (80.7%) displayed a normal chromosome constitution. Three cases of chromosome non-disjunction (2.8%) were found, whereas seven oocytes (6.7%) presented extra single chromatids. In addition, 12 oocytes (11.5%) showed balanced pre-division of one pair of sister chromatids. Although this phenomenon was not classified as aneuploidy, it could lead to aneuploidy at anaphase II. Abnormalities were observed in all the targetted chromosomes. The present data confirm that both whole chromosome non- disjunction and premature chromatid separation constitute the two major mechanisms of aneuploidy in human female meiosis.
Key words: aneuploidy/double-labelling/FISH/oocyte/polar body
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