Sakkas D, Urner F, Bianchi P G, Bizzaro D, Wagner I, Jaquenoud N, Manicardi G, Campana A
Clinic of Infertility and Gynaecological Endocrinology--WHO Collaborating Centre, University Hospital of Geneva, Switzerland.
Hum Reprod. 1996 Apr;11(4):837-43. doi: 10.1093/oxfordjournals.humrep.a019263.
In this study we investigated whether morphology and chromatin anomalies in human spermatozoa can influence fertilization after intracytoplasmic sperm injection (ICSI). We examined unfertilized oocytes, using the fluorochrome Hoechst 33342, to determine whether a relationship exists between failure of fertilization and sperm chromatin quality. Sperm chromatin packaging quality was assessed using the chromomycin A3 (CMA3) fluorochrome, and the presence of DNA damage in spermatozoa, using in-situ nick translation, Normal males present sperm parameters with a normal morphology of > 20%, CMA3 fluorescence of < 30% and exhibit endogenous nicks in < 10% of their spermatozoa. When patients were separated according to these values no difference was observed in their fertilization rates after ICSI. When the unfertilized ICSI oocytes were examined, we found that patients with CMA3 fluorescence of <30% and nicks in < 10% of their spermatozoa had only 17.5 and 21.6% respectively of their unfertilized oocytes containing spermatozoa that remained condensed. In contrast, patients with higher CMA3 and nick values had a significantly higher number, 41.2 and 48.9%, of their unfertilized oocytes containing condensed spermatozoa. sperm morphology did not show any such pattern. The percentage of spermatozoa which had initiated decondensation in unfertilized oocytes was not influenced by morphology, CMA3 fluorescence or nicks. In light of these results we postulate that poor chromatin packaging and/or damaged DNA may contribute to failure of sperm decondensation after ICSI and result in failure of fertilization.
在本研究中,我们调查了人类精子的形态和染色质异常是否会影响卵胞浆内单精子注射(ICSI)后的受精情况。我们使用荧光染料Hoechst 33342检查未受精的卵母细胞,以确定受精失败与精子染色质质量之间是否存在关联。使用放线菌素A3(CMA3)荧光染料评估精子染色质包装质量,并通过原位缺口平移检测精子中的DNA损伤情况。正常男性的精子参数为:形态正常率>20%,CMA3荧光率<30%,且<10%的精子存在内源性缺口。根据这些值对患者进行分组后,未观察到他们ICSI后的受精率有差异。当检查未受精的ICSI卵母细胞时,我们发现CMA3荧光率<30%且<10%的精子存在缺口的患者,其未受精卵母细胞中含有仍呈浓缩状态精子的比例分别仅为17.5%和21.6%。相比之下,CMA3和缺口值较高的患者,其未受精卵母细胞中含有浓缩精子的比例显著更高,分别为41.2%和48.9%。精子形态未显示出任何此类模式。未受精卵母细胞中已开始解聚的精子百分比不受形态、CMA3荧光或缺口的影响。鉴于这些结果,我们推测染色质包装不良和/或DNA损伤可能导致ICSI后精子解聚失败并导致受精失败。