Weissenberg R, Aviram A, Golan R, Lewin L M, Levron J, Madgar I, Dor J, Barkai G, Goldman B
Andrology Unit, Sheba Medical Center, Tel Hashomer, Ramat Gan, Israel.
Mol Hum Reprod. 1998 Jan;4(1):61-6. doi: 10.1093/molehr/4.1.61.
Routine semen analysis in an infertile patient revealed severe teratospermia associated with malformation of head and tail in 100% of the sperm cells. Flow cytometry and fluorescence in-situ hybridization (FISH) were shown to supplement routine semen analysis by providing information on the sperm chromatin. Using flow cytometry, propidium iodide-stained spermatozoa from the same sperm sample were compared with a normal reference pool, and with human lymphocytes. The results point to a population of diploid sperm cells rather than to mature haploid spermatozoa. Numerical chromosomal abnormalities of the spermatozoa were subsequently evaluated using FISH. A total of 1000 sperm cells were scored for X and Y chromosomes, and an additional 1128 sperm cells for chromosome 18. Aneuploidy of chromosomes X and Y was revealed in 96.9% of the cells and of chromosome 18 in 90.3% of the cells. Non-disjunction of chromosome X and Y in meiosis I and II occurred in 54.8 and 2.7% of the sperm cells respectively. Non-disjunction in both meiosis I and II occurred in 39.4% of the sperm cells. A normal haploid pattern for chromosomes X and Y was observed in only 3.1%, and for chromosome 18 in 9.7%, of the cells. Using three colour FISH for the sex chromosomes and for chromosome 18, diploidy was demonstrated in 19.4% of 500 sperm cells and aneuploidy in virtually all sperm cells (99.2%). The use of flow cytometry and FISH in cases where genetic and developmental chromatin abnormalities are suspected is a valuable adjunct to other available techniques, and can guide the clinicians to decide which samples are unsuitable for intracytoplasmic injection.
对一名不育患者进行的常规精液分析显示，100%的精子细胞存在严重畸形精子症，伴有头部和尾部畸形。流式细胞术和荧光原位杂交（FISH）被证明可通过提供精子染色质信息来补充常规精液分析。使用流式细胞术，将来自同一精液样本的碘化丙啶染色精子与正常参考库以及人类淋巴细胞进行比较。结果表明存在一群二倍体精子细胞而非成熟的单倍体精子。随后使用FISH评估精子的染色体数目异常。对总共1000个精子细胞进行X和Y染色体评分，另外对1128个精子细胞进行18号染色体评分。96.9%的细胞显示X和Y染色体非整倍体，90.3%的细胞显示18号染色体非整倍体。减数分裂I和II中X和Y染色体的不分离分别发生在54.8%和2.7%的精子细胞中。减数分裂I和II均发生不分离的情况出现在39.4%的精子细胞中。仅3.1%的细胞观察到X和Y染色体的正常单倍体模式，9.7%的细胞观察到18号染色体的正常单倍体模式。使用针对性染色体和18号染色体的三色FISH，在500个精子细胞中有19.4%显示二倍体，几乎所有精子细胞（99.2%）显示非整倍体。在怀疑存在遗传和发育染色质异常的情况下，使用流式细胞术和FISH是对其他现有技术的有价值补充，并且可以指导临床医生决定哪些样本不适合进行胞浆内注射。
