Sequential multiple probe fluorescence in-situ hybridization analysis of human oocytes and polar bodies by combining centromeric labelling and whole chromosome painting.
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.
Anahory
T
T
Service de Génétique Moléculaire et Chromosomique, CHU Arnaud de Villeneuve, 34295 Montpellier, Cedex 5, France.
Andréo
B
B
Régnier-Vigouroux
G
G
Soulie
J P
JP
Baudouin
M
M
Demaille
J
J
Pellestor
F
F
D002503:Centromere / Q000235:genetics*; D020223:Chromosome Painting / Q000379:methods*; D002877:Chromosomes, Human / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics*; D005260:Female; D006801:Humans; D008677:Metaphase; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism*
D015342:DNA Probes