Martin R H, Spriggs E, Rademaker A W
Department of Medical Genetics, University of Calgary, Alberta, Canada.
Biol Reprod. 1996 Feb;54(2):394-8. doi: 10.1095/biolreprod54.2.394.
Aneuploidy and diploidy frequencies for chromosomes 1, 12, X, and Y were assessed in 225,846 sperm from 10 normal men. Results from 5 of the men have previously been reported. Multicolor fluorescence in situ hybridization (FISH) was used to control for lack of probe hybridization and to distinguish diploidy from disomy. A minimum of 10,000 sperm per donor were evaluated for each chromosome. Sperm were considered disomic if two fluorescent signals were separated by a distance of a minimum of one signal domain. The mean frequencies of disomic sperm for chromosomes 1 and 12 were 0.11% (range 0.05-0.18%) and 0.16% (range 0.10-0.25%), respectively. The means for the sex chromosomal aneuploidies were 0.07% XX, 0.18% YY, and 0.16% XY, totaling 0.42% for all sex chromosomes (range 0.23-0.71%). The incidence of disomic sperm for the sex chromosomes was significantly increased compared to the frequency for the autosomes, corroborating results obtained from studies of sperm karyotypes and spontaneous abortions. The mean frequencies of single X- and Y-bearing sperm were 50.1% and 49.0%, respectively--not significantly different from 50%. The mean frequency of diploid sperm was 0.16% (0.06-0.42%). Interdonor heterogeneity was found to exist for disomy 1, XX, YY, and diploidy, suggesting significant variation among normal men. Comparison of these FISH results to our historical sperm karyotypes demonstrated that the sex ratios and disomy frequencies for chromosomes 1 and X were similar. However, there was a significantly increased frequency of disomic sperm for chromosomes 12, YY, and XY in FISH data compared with sperm karyotypes. In general, FISH data agreed quite well with values from sperm karyotyping, including the increased frequency of sex chromosomal aneuploidy compared with autosomal aneuploidy in sperm. Multicolor FISH analysis permits an accurate distinction between disomic and diploid sperm and allows analysis of large sample sizes. This powerful technology may be useful for future studies of potential environmental and occupational mutagens.
对10名正常男性的225,846条精子评估了1号、12号、X和Y染色体的非整倍体和二倍体频率。其中5名男性的结果先前已报道。采用多色荧光原位杂交（FISH）技术来控制探针杂交失败的情况，并区分二倍体和双体。每个供体的每条染色体至少评估10,000条精子。如果两个荧光信号之间的距离至少为一个信号域，则精子被认为是双体。1号和12号染色体双体精子的平均频率分别为0.11%（范围0.05 - 0.18%）和0.16%（范围0.10 - 0.25%）。性染色体非整倍体的平均值为XX 0.07%、YY 0.18%、XY 0.16%，所有性染色体总计0.42%（范围0.23 - 0.71%）。与常染色体相比，性染色体双体精子的发生率显著增加，这与精子核型和自然流产研究结果一致。含单条X和Y精子的平均频率分别为50.1%和49.0%，与50%无显著差异。二倍体精子的平均频率为0.16%（0.06 - 0.42%）。发现1号、XX、YY染色体双体和二倍体存在供体间异质性，表明正常男性之间存在显著差异。将这些FISH结果与我们既往的精子核型进行比较，结果显示1号和X染色体的性别比例和双体频率相似。然而，与精子核型相比，FISH数据中12号、YY和XY染色体双体精子的频率显著增加。总体而言，FISH数据与精子核型分析结果相当吻合，包括精子中性染色体非整倍体频率高于常染色体非整倍体频率。多色FISH分析能够准确区分双体和二倍体精子，并允许对大样本进行分析。这项强大的技术可能对未来潜在环境和职业诱变剂的研究有用。
