Detection of aneuploidy in human interphase spermatozoa by fluorescence in situ hybridization (FISH)

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104

To understand the mechanisms that affect aneuploidy, fluorescence in situ hybridization (FISH), using chromosome-specific centromeric probes, was employed to screen a large population of human sperm for numerical errors. To determine the true rate of disomy for chromosomes 1, 12, 15, and 18, two-color FISH was performed, and for the gonosomes, three-color FISH. The use of multiple, differently colored probes allows one to distinguish a true disomic sperm from a diploid cell. A minimum of 10,000 sperm nuclei from each of five donors was scored per set of centromeric probes, giving a total of 165,330 sperm nuclei. The disomy frequencies for autosomes 1, 12, 15, and 18 were found to be similar, with a mean of 0.10% (range, 0.05%–0.16%) for chromosome 1, 0.16% (0.10%–0.25%) for chromosome 12, 0.11% (0.07%–0.20%) for chromosome 15, and 0.11% (0.08%–0.17%) for chromosome 18. For the sex chromosomes, the mean frequency of disomy was found to be 0.43% (range, 0.23%–0.71%), with XX disomy accounting for 0.07% (0.03%–0.10%), YY disomy 0.21% (0.10%–0.43%), and XY disomy 0.15% (0.08%–0.24%). The incidence of disomic sperm for the sex chromosomes was significantly increased, compared to the frequency of disomy for the autosomes (χ² = 218.61 P < 0.0001). Diploidy was observed in 0.05%-0.47% of the sperm nuclei counted. Interdonor heterogeneity for disomy frequencies was found to exist for the sex chromosomes and for chromosomes 1 and 15, suggesting significant variation among normal men.

“…1994. the majority of these studies was performed using one-color FISH Martin (1993) 0.06 0.04 0.03 Robbins et al (1993) 0.14 0.06 Martin et al (1993) 0.14 0.17…”

Section: Discussionmentioning

confidence: 99%

Aneuploidy in human sperm: results of two- and three-color fluorescence in situ hybridization using centromeric probes for chromosomes 1, 12, 15, 18, X, and Y

Spriggs

Rademaker

Martin

1995

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104

“…Sperm chromosome complements can be assessed by two different methods; human sperm karyotyping using the hamster oocyte-human sperm fusion system (Rudak et al, 1978;Martin et al, 1983;Brandriff et al. 1985) and by fluorescence in situ hybridization (FISH) using chromosomespecific DNA probes (Wyrobek et al, 1990;Martin et al, 1993Martin et al, , 1996Williams et al, 1993). Each of these methods has advan tages and disadvantages.…”

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confidence: 99%

Analysis of human sperm karyotypes in testicular cancer patients before and after chemotherapy

Martin¹,

Ernst

Rademaker

et al. 1997

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Sperm karyotype analysis was performed on testicular cancer patients before and after treatment with BEP (bleomycin, etoposide, and cisplatin). A total of 788 sperm chromosome complements was studied, 236 before chemotherapy (CT) and 552 post-CT. There was no significant difference in the total frequency of sperm chromosomal abnormalities pre-CT (10.2%) compared to post-CT (10.7 %). Similarly, there were no significant differences in the frequencies of numerical abnormalities (2.5% pre-CT vs. 2.4% post-CT) or structural abnormalities (6.4% pre-CT vs. 7.4% post-CT). The percentage of X-bearing sperm was also not significantly different before (46.3%) and after CT (50.1%). The results in cancer patients were not significantly different from those in control donors. This study corroborates results from our previous analysis of these same men using multicolor fluorescence in situ hybridization for assessment of aneuploidy for chromosomes 1, 12, X, Y, and XY. Together, these two studies suggest that the sperm of men receiving BEP chemotherapy are not at increased risk of chromosomal abnormalities two or more years after treatment.

“…1991: Jackson-Cook andHaller, 1991;Han et al. 1992: Holmes andMartin et al, 1993;Robbins et al, 1993;Guttenbach et al" 1994;Miharu et al. 1994) or two-color FISH (Goldman et al.…”

mentioning

confidence: 99%

Meiotic segregation of the X and Y chromosomes and chromosome 1 analyzed by three-color FISH in human interphase spermatozoa

Chevret

Rousseaux

Monteil

et al. 1995

Meiotic segregation of the X and Y chromosomes and chromosome 1 was analyzed by three-color fluorescence in situ hybridization (FISH) in 94,575 human interphase spermatozoa from four control subjects. More than 99% of the sperm cells were labeled. The proportions of X- and Y-bearing sperm were estimated to be 49.83% and 48.30%, respectively. The disomy rates were 0.04%, 0.009%, and 0.20% for the X and Y chromosomes and chromosome 1, respectively. Hyperhaploidy with an extra gonosome was found in 0.34% of spermatozoa, due to nondisjunction during meiosis I. The frequency of diploidy was 0.11 % at meiosis I and 0.036% at meiosis II. Cohybridization of one autosomal and two gonosomal probes, in three-color FISH in interphase spermatozoa, seems to accurately discriminate diploidies from disomies, as well as the meiotic origin of gonosomal aneuploidies in sperm cells.