Lolis D, Georgiou I, Syrrou M, Zikopoulos K, Konstantelli M, Messinis I
Medical School, University of Ioannina.
Int J Androl. 1996 Feb;19(1):23-7. doi: 10.1111/j.1365-2605.1996.tb00429.x.
Mature mammalian spermatozoa have a compact and stable nuclear structure conferred by protamines instead of histones, which are present in all other cellular types. Chromomycin A3 (CMA3) is a useful tool for the detection of protamine deficiency in sperm chromatin. The purpose of this study was to correlate the percentage of spermatozoa staining positively for CMA3 with sperm parameters and in-vitro fertilization of human oocytes. Spermatozoa were collected from 56 fertile and 18 infertile men, and washed twice in PBS, fixed in two changes of methanol : acetic acid (3 : 1 v : v) spread on rinsed slides treated with APES and dried. Twenty-four of the semen samples were subjected to both Percoll and swim-up, and were stained subsequently with CMA3. CMA3-stained spermatozoa were expressed as a percentage in a count of 200 spermatozoa. A substantial variation in the percentage of CMA3-stained cells was observed in ejaculated human spermatozoa, varying between 8% and 77%. A strong negative correlation (r = -0.64, p < 0.001) was found between sperm count and the percentage of CMA3-stained spermatozoa. No correlation was found between CMA3-stained spermatozoa and their motility, while excessive sperm morphological abnormalities were related positively to CMA3-staining. Spermatozoa in samples exhibiting low (8-62%) CMA3-staining had significantly higher fertilizing rates in vitro than did samples exhibiting high (49-77%) CMA3-staining. The mean percentage of CMA3-stained spermatozoa after swim-up or Percoll preparation (26% vs 31%) did not differ significantly. These results demonstrate a close relationship between CMA3-staining, fertilization and sperm count, and suggest potential application of this marker for the prediction of sperm quality and fertilizing capacity.
成熟的哺乳动物精子具有由鱼精蛋白而非组蛋白赋予的紧密且稳定的核结构，而组蛋白存在于所有其他细胞类型中。嗜铬霉素A3（CMA3）是检测精子染色质中鱼精蛋白缺乏的有用工具。本研究的目的是将CMA3染色阳性的精子百分比与精子参数以及人类卵母细胞的体外受精相关联。从56名有生育能力的男性和18名不育男性中收集精子，在PBS中洗涤两次，用甲醇:乙酸（3:1 v:v）固定两次，铺展在用APES处理并冲洗过的载玻片上，然后干燥。24份精液样本经过Percoll和上游法处理，随后用CMA3染色。CMA3染色的精子以200个精子计数中的百分比表示。在射出的人类精子中观察到CMA3染色细胞百分比存在很大差异，在8%至77%之间变化。精子计数与CMA3染色精子的百分比之间发现强烈的负相关（r = -0.64，p < 0.001）。未发现CMA3染色的精子与其活力之间存在相关性，而过多的精子形态异常与CMA3染色呈正相关。CMA3染色低（8 - 62%）的样本中的精子在体外的受精率显著高于CMA3染色高（49 - 77%）的样本。上游法或Percoll制备后CMA3染色精子的平均百分比（26%对31%）无显著差异。这些结果表明CMA3染色、受精和精子计数之间存在密切关系，并表明该标志物在预测精子质量和受精能力方面的潜在应用。
