Huszar G, Vigue L, Oehninger S
Department of Obstetrics and Gynecology, Yale School of Medicine, New Haven, Connecticut 06510-0863.
Fertil Steril. 1994 Jan;61(1):136-42. doi: 10.1016/s0015-0282(16)56466-8.
To examine the clinical significance of the increased sperm cytoplasmic content that is due to a fault of spermatogenesis, we have further studied the relationship between increased sperm creatine kinase (CK) concentrations and diminished fertilizing potential in men. In the present work, we used CK immunocytochemistry of human sperm-hemizona (HZ) complexes to examine whether the distribution of mature (clear heads), intermediate (sperm heads with light stippling), and immature (heads with heavy stippling or with solid CK staining) spermatozoa bound to the HZ would follow the incidence of these sperm in the samples tested, or if there is a preferential binding by the mature sperm.
Two pairs of HZ were exposed to washed semen and to their swim-up sperm fractions. The sperm and sperm-HZ complexes were treated with a CK antibody followed by horseradish peroxidase immunostaining, and the sperm were evaluated for maturity.
Men presenting for fertility evaluation were studied in two university-based andrology laboratories.
The binding of the HZ was selective for mature sperm as indicated by the incidence of intermediate and immature sperm in washed semen versus bound to the HZ (intermediate: 20.0% versus 1.4%; immature: 7.6% versus 0.5% [mean +/- SEM]) or in swim-up sperm fractions versus the HZ (intermediate: 18.7% versus 3.4%; immature: 2.5% versus 0.2%). The binding was almost exclusive to normal sperm (96.4% to 98.1%) whether the HZ were exposed to washed semen or swim-up fractions in spite of the five to ten times higher incidence of intermediate and immature sperm.
Mature sperm selectively bind to the zona. We suggest that spermatozoa with immature CK-staining patterns are deficient in the site(s) of oocyte recognition and binding.
为了研究由于精子发生缺陷导致的精子细胞质含量增加的临床意义，我们进一步研究了男性精子肌酸激酶（CK）浓度升高与受精潜能降低之间的关系。在本研究中，我们使用人精子 - 半透明带（HZ）复合物的CK免疫细胞化学方法，来检测与HZ结合的成熟（头部清亮）、中间型（头部有轻度点状染色）和未成熟（头部有重度点状染色或CK呈实心染色）精子的分布情况是否与测试样本中这些精子的发生率一致，或者成熟精子是否存在优先结合现象。
将两对HZ分别与洗涤后的精液及其上游精子部分进行接触。精子和精子 - HZ复合物先用CK抗体处理，然后进行辣根过氧化物酶免疫染色，并评估精子的成熟度。
在两个大学附属的男科实验室对前来进行生育力评估的男性进行研究。
HZ的结合对成熟精子具有选择性，这体现在洗涤后的精液中与结合到HZ上的中间型和未成熟精子的发生率上（中间型：20.0% 对1.4%；未成熟型：7.6% 对0.5% [平均值±标准误]），或者在上游精子部分与HZ的结合情况中（中间型：18.7% 对3.4%；未成熟型：2.5% 对0.2%）。无论HZ是与洗涤后的精液还是上游精子部分接触，结合几乎都只发生在正常精子上（96.4%至98.1%），尽管中间型和未成熟精子的发生率要高出五到十倍。
成熟精子选择性地结合到透明带上。我们认为，具有未成熟CK染色模式的精子在卵母细胞识别和结合位点上存在缺陷。
