Van Hummelen P, Lowe X R, Wyrobek A J
Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, CA 94550, USA.
Hum Genet. 1996 Nov;98(5):608-15. doi: 10.1007/s004390050268.
Both structural and numerical chromosome aberrations in sperm represent important categories of paternally transmitted genetic damage. Therefore, a new multiprobe fluorescence in situ hybridization (FISH) method, using DNA probes for three targets (centromere and telomere of chromosome 1, centromere of chromosome 8), was developed to detect human sperm carrying three types of chromosomal defects: (1) terminal duplications or deletions in chromosome 1p, (2) aneuploidy involving chromosomes 1 or 8, and (3) diploidy. Baseline frequencies were determined for three healthy donors who had been previously evaluated for sperm cytogenetics by the human-sperm/hamster-oocyte cytogenetic technique (hamster technique). Among approximately 120,000 sperm analyzed by the new FISH method, the average baseline frequencies of sperm carrying telomeric duplications and deletions of 1p were 3.2 +/- 1.9 and 2.9 +/- 3.6 per 10(4), respectively. Diploid sperm was found in an average frequency of 6.6 +/- 4.0 per 10(4). Average frequencies of disomic sperm for chromosomes 1 or 8 were 1.7 +/- 2.2 and 1.9 +/- 2.3 per 10(4), respectively. Inter-individual differences were observed for deletions of 1p but not for the other sperm phenotypes. A good correlation was obtained between the frequencies of sperm with structural chromosome aberrations detected with the new assay and the frequency of sperm carrying premeiotic or meiotic cytogenetic damage detected with the hamster technique. The observed levels of numerical aberrations with the new FISH assay were within range of the baseline frequencies reported by the hamster technique. The newly developed FISH assay has promising applications in genetic, clinical, physiological and toxicological studies.
精子中的染色体结构和数目畸变是父系遗传损伤的重要类型。因此，开发了一种新的多探针荧光原位杂交（FISH）方法，使用针对三个靶点（1号染色体的着丝粒和端粒、8号染色体的着丝粒）的DNA探针，以检测携带三种染色体缺陷类型的人类精子：（1）1号染色体短臂的末端重复或缺失；（2）涉及1号或8号染色体的非整倍体；（3）二倍体。对三名健康供体的基线频率进行了测定，这三名供体此前已通过人精子/仓鼠卵母细胞细胞遗传学技术（仓鼠技术）进行了精子细胞遗传学评估。在用新的FISH方法分析的约120,000个精子中，携带1号染色体短臂端粒重复和缺失的精子的平均基线频率分别为每10⁴个3.2±1.9和2.9±3.6。二倍体精子的平均频率为每10⁴个6.6±4.0。1号或8号染色体二体精子的平均频率分别为每10⁴个1.7±2.2和1.9±2.3。观察到1号染色体短臂缺失存在个体间差异，但其他精子表型不存在个体间差异。新检测方法检测到的具有染色体结构畸变的精子频率与仓鼠技术检测到的携带减数分裂前或减数分裂细胞遗传学损伤的精子频率之间存在良好的相关性。新的FISH检测方法观察到的数目畸变水平在仓鼠技术报告的基线频率范围内。新开发的FISH检测方法在遗传、临床、生理和毒理学研究中有广阔的应用前景。
