Francavilla S, Gabriele A, Romano R, Gianaroli L, Ferraretti A P, Francavilla F
Department of Internal Medicine, University of L'Aquila, Bologna, Italy.
Fertil Steril. 1994 Dec;62(6):1226-33. doi: 10.1016/s0015-0282(16)57190-8.
To determine whether the sperm-zona pellucida (ZP) binding is related to the presence of immunoreactivity for proacrosin and acrosin and/or to the proteolytic activity of acrosin.
The Andrology Clinic, University of L'Aquila, Italy.
Thirty-five infertile couples and 15 men of proven fertility.
Salt-stored unfertilized oocytes after IVF were inseminated with a mixture of equal numbers of test and fertile donor sperm after swim-up selection, respectively labeled with fluorescein or rhodamine. Total acrosin activity, the percent of sperm immunostained for proacrosin and acrosin, and the percent of sperm with oval heads were evaluated in fresh semen and after swim-up selection.
Three zonae were used for each patient and sperm-ZP binding ratio (ZP ratio) was calculated as the total number of test sperm bound to ZP divided by the total number of control sperm bound to ZP. The ZP ratio was compared with the percent of live sperm immunostained for proacrosin and acrosin, with total acrosin activity and with the percent of sperm with oval heads after swim-up selection.
The sperm-ZP binding ratio was correlated positively with the percent of sperm immunostained for proacrosin and acrosin and with the percent of sperm with oval heads, but not with acrosin activity. Ejaculates with immunostaining for proacrosin and acrosin > or = 55% of sperm (the mean value of infertile couples) and acrosin activity < 24 muIU/10(6) sperm (the mean value of infertile couples) (n = 8) had a ZP ratio > or = 0.5, whereas ejaculates with low immunostaining for proacrosin and low acrosin activity (n = 12) had a ZP ratio < 0.5. The difference between the ZP ratio of the two groups was highly significant.
The ZP binding is related to the immunocytochemical presence of proacrosin and acrosin but not to the proteolytic activity of acrosin in human sperm, suggesting that egg recognition and proteolytic activity are independent functions of proacrosin and acrosin.
确定精子与透明带（ZP）的结合是否与前顶体蛋白酶和顶体蛋白酶的免疫反应性存在有关，和/或与顶体蛋白酶的蛋白水解活性有关。
意大利拉奎拉大学男科诊所。
35对不育夫妇和15名已证实具有生育能力的男性。
体外受精后经盐储存的未受精卵母细胞，分别用等量经上浮法筛选的受试精子和可育供体精子混合物进行授精，受试精子和供体精子分别用荧光素或罗丹明标记。对新鲜精液和上浮法筛选后的精液评估顶体蛋白酶总活性、前顶体蛋白酶和顶体蛋白酶免疫染色阳性的精子百分比，以及头部呈椭圆形的精子百分比。
每位患者使用三个透明带，精子 - ZP结合率（ZP比率）计算为与ZP结合的受试精子总数除以与ZP结合的对照精子总数。将ZP比率与前顶体蛋白酶和顶体蛋白酶免疫染色阳性的活精子百分比、顶体蛋白酶总活性以及上浮法筛选后头部呈椭圆形的精子百分比进行比较。
精子 - ZP结合率与前顶体蛋白酶和顶体蛋白酶免疫染色阳性的精子百分比以及头部呈椭圆形的精子百分比呈正相关，但与顶体蛋白酶活性无关。前顶体蛋白酶和顶体蛋白酶免疫染色阳性的精子≥55%（不育夫妇的平均值）且顶体蛋白酶活性<24 mIU/10⁶精子（不育夫妇的平均值）的射精样本（n = 8），其ZP比率≥0.5，而前顶体蛋白酶免疫染色阳性率低且顶体蛋白酶活性低的射精样本（n = 12），其ZP比率<0.5。两组的ZP比率差异非常显著。
在人类精子中，ZP结合与前顶体蛋白酶和顶体蛋白酶的免疫细胞化学存在有关，但与顶体蛋白酶的蛋白水解活性无关，这表明卵子识别和蛋白水解活性是前顶体蛋白酶和顶体蛋白酶的独立功能。
