Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479 Poznan, Poland.
Reprod Biol. 2013 Sep;13(3):209-20. doi: 10.1016/j.repbio.2013.06.002. Epub 2013 Jul 18.
The presence of reciprocal chromosome translocations (RCTs), as well as sperm chromatin disturbances, is known to exert negative influence on male fertility. The aim of this study was to identify an association between chromosome structural rearrangements in male RCT carriers and sperm seminological parameters (concentration, motility, morphology), chromatin status (fragmentation and maturity), meiotic segregation pattern and observed chromosomal hyperhaploidy. Sperm samples originated from ten male RCT carriers with reproductive failure/success. TUNEL assay (DNA fragmentation) and chromomycin A3 (CMA3)/aniline blue (AB) staining (chromatin maturity) were used to analyze sperm chromatin status while fluorescent in situ hybridization (FISH) was applied to observe meiotic segregation patterns and hyperhaploidy in spermatozoa. We found that the mean level of sperm DNA fragmentation in the RCT carrier group (18.0 ± 11.9%) was significantly higher (p=0.0006) than the mean of the control group (7.5 ± 4.3%). There was no correlation observed between sperm DNA fragmentation levels (5.6-38.0%) and the frequency of genetically normal/balanced gametes (34.3-62.4%), sperm seminological quality or revealed reproductive failure. In contrast, a correlation between the frequencies of genetically normal/balanced spermatozoa and of gametes with mature chromatin was observed (CMA3: R=0.4524, p=0.2604; AB: R=0.5238, p=0.1827). A statistically significant increase in the hyperhaploidy level of selected chromosomes in all analyzed RCT carriers was documented but was not correlated to sperm seminology or fertility status. Further evaluation and additional assays toward sperm chromatin quality assessment in RCT carriers is suggested to explain the complexity of genomic structural rearrangements and its possible relevance to reproductive success or failure.
染色体相互易位（RCT）的存在以及精子染色质紊乱已知会对男性生育力产生负面影响。本研究的目的是确定男性 RCT 携带者的染色体结构重排与精子精液参数（浓度、活力、形态）、染色质状态（碎片化和成熟度）、减数分裂分离模式和观察到的染色体超二倍体之间的关联。精子样本来自 10 名具有生殖失败/成功的男性 RCT 携带者。TUNEL 检测（DNA 碎片化）和染色质 A3（CMA3）/苯胺蓝（AB）染色（染色质成熟度）用于分析精子染色质状态，而荧光原位杂交（FISH）用于观察精子减数分裂分离模式和超二倍体。我们发现，RCT 携带者组的精子 DNA 碎片化平均水平（18.0±11.9%）明显高于对照组（7.5±4.3%）（p=0.0006）。未观察到精子 DNA 碎片化水平（5.6-38.0%）与正常/平衡配子的频率（34.3-62.4%）、精子精液质量或显示的生殖失败之间存在相关性。相反，观察到正常/平衡精子和成熟染色质的配子的频率之间存在相关性（CMA3：R=0.4524，p=0.2604；AB：R=0.5238，p=0.1827）。在所有分析的 RCT 携带者中，都记录到所选染色体超二倍体水平的统计学显著增加，但与精子精液学或生育状态无关。建议对 RCT 携带者的精子染色质质量进行进一步评估和额外检测，以解释基因组结构重排的复杂性及其对生殖成功或失败的可能相关性。
