Baumgartner A, Van Hummelen P, Lowe X R, Adler I D, Wyrobek A J
Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, California 94550, USA.
Environ Mol Mutagen. 1999;33(1):49-58. doi: 10.1002/(sici)1098-2280(1999)33:1<49::aid-em6>3.0.co;2-f.
A pair of multicolor FISH assays (X-Y-21 and A-M-16) was developed for human sperm to simultaneously measure sex ratios; aneuploidies involving chromosomes 1, 16, 21, X, and Y; meiotic diploidies; and structural aberrations involving chromosome 1p. Sex ratios in sperm were not significantly different from unity among healthy men. Baseline frequencies of disomic sperm for chromosomes 1, 8, and 21 were similar (6.7 per 10(4) sperm, 95% CI of 5.6-8.1), suggesting that among these three chromosomes, chromosome 21 was not especially prone to nondisjunction. Frequencies of disomy 16 sperm were significantly lower, however (3.5 per 10(4) sperm, 95% CI of 2.0-6.2; P < 0.02). The baseline frequencies of sperm disomy by FISH for chromosomes 16 and 21 were validated against aneuploidy data obtained by the hamster-egg technique for human sperm cytogenetics. The frequencies of X-X, Y-Y, X-Y ("Klinefelter") sperm and sex-null ("Turner") sperm were 5.5, 5.1, 5.5, and 7.8 per 10(4) sperm, respectively. For chromosomes 16 and 21, the frequencies of nullisomic and disomic sperm were similar, suggesting that gain and loss events occurred symmetrically. However, more gain than loss was reported for chromosomes 1, X, and Y. The frequency of MI and MII diploid sperm (with flagella) was approximately 12 per 10(4) (range 8.3-16.7 per 10(4) sperm). Based on flagella data, the frequency of somatic cells in the semen was estimated to be approximately 1.8 per 10(4) sperm. Loss or gain of a portion of chromosome-arm 1p occurred in 5.5 per 10(4) sperm, and the percentage of sperm carrying structural aberrations within the haploid genome as calculated from FISH (1.4%), was similar to that obtained with the hamster-egg technique. These complementary sperm FISH assays have promising applications in studies of chromosomally abnormal sperm after exposure to occupational, medical, and environmental toxicants.
开发了一对用于人类精子的多色荧光原位杂交检测方法（X - Y - 21和A - M - 16），以同时测量性别比例；涉及1号、16号、21号、X和Y染色体的非整倍体；减数分裂二倍体；以及涉及1号染色体短臂的结构畸变。健康男性精子中的性别比例与1无显著差异。1号、8号和21号染色体的二体精子基线频率相似（每10⁴个精子中有6.7个，95%置信区间为5.6 - 8.1），这表明在这三条染色体中，21号染色体并非特别容易发生不分离。然而，16号染色体二体精子的频率显著较低（每10⁴个精子中有3.5个，95%置信区间为2.0 - 6.2；P < 0.02）。通过荧光原位杂交检测到的16号和21号染色体二体精子的基线频率，与通过仓鼠卵技术获得的人类精子细胞遗传学非整倍体数据进行了验证。X - X、Y - Y、X - Y（“克兰费尔特综合征”）精子和无性染色体（“特纳综合征”）精子的频率分别为每10⁴个精子5.5个、5.1个、5.5个和7.8个。对于16号和21号染色体，单体和二体精子的频率相似，这表明增加和缺失事件对称发生。然而，据报道，1号、X和Y染色体增加的情况多于缺失。MI和MII二倍体精子（有鞭毛）的频率约为每10⁴个中有12个（范围为每10⁴个精子8.3 - 16.7个）。根据鞭毛数据，精液中体细胞的频率估计约为每10⁴个精子中有1.8个。1号染色体短臂的部分缺失或增加发生在每10⁴个精子中有5.5个，通过荧光原位杂交计算得出的单倍体基因组中携带结构畸变的精子百分比（1.4%），与通过仓鼠卵技术获得的结果相似。这些互补的精子荧光原位杂交检测方法在研究接触职业、医疗和环境毒物后染色体异常精子方面具有广阔的应用前景。
