Oehninger S, Mahony M C, Swanson J R, Hodgen G D
Jones Institute for Reproductive Medicine, Department of Obstetrics and Gynecology, Eastern Virginia Medical School, Norfolk 23507.
Mol Reprod Dev. 1993 May;35(1):57-61. doi: 10.1002/mrd.1080350110.
The objective of this prospective study was to evaluate the specificity of human sperm/zona pellucida interaction under hemizona assay (HZA) conditions in experiments with gametes from the same and different species. Human, cynomolgus monkey and hamster oocytes were used after salt-storage. Oocytes were bisected into matching hemizonae by micromanipulation and used in the HZA. Semen was obtained from healthy men (donors) and male cynomolgus monkeys and prepared by wash and swim-up. Sperm binding to matching hemizonae was assessed (tight binding) after 4-h coincubation in the HZA in homologous and interspecies experiments. Acrosome reaction was evaluated in the sperm droplets using FITC-PSA and on the hemizonae using the T-6 monoclonal antibody. On human hemizonae, the number of tightly bound sperm for human and monkey were 93.2 +/- 15.8 and 3.9 +/- 1.3, respectively (P < 0.001). On monkey hemizonae, the number of tightly bound sperm for monkey and human were 126.0 +/- 34.8 and 2.8 +/- 1.6, (P = 0.02) respectively. On hamster hemizonae, there was negligible binding of human and monkey sperm. There was a significantly higher incidence of acrosome reacted sperm on the zona pellucida in homologous compared to heterologous experiments. These results demonstrate a high species-specificity of human gamete functions under HZA conditions, providing further support for the use of this bioassay in infertility and contraception testing.
这项前瞻性研究的目的是在半透明带分析(HZA)条件下,评估来自同一物种和不同物种的配子实验中人类精子/透明带相互作用的特异性。盐储存后使用人类、食蟹猴和仓鼠的卵母细胞。通过显微操作将卵母细胞切成匹配的半透明带,并用于HZA。精液取自健康男性(供体)和雄性食蟹猴,经洗涤和上浮法制备。在同源和种间实验的HZA中共同孵育4小时后,评估精子与匹配半透明带的结合情况(紧密结合)。使用异硫氰酸荧光素标记的前列腺特异性抗原(FITC-PSA)在精子液滴中评估顶体反应,并使用T-6单克隆抗体在半透明带上评估顶体反应。在人类半透明带上,人类和猴子紧密结合的精子数量分别为93.2±15.8和3.9±1.3(P<0.001)。在猴子半透明带上,猴子和人类紧密结合的精子数量分别为126.0±34.8和2.8±1.6(P = 0.02)。在仓鼠半透明带上,人类和猴子精子的结合可以忽略不计。与异种实验相比,同源实验中透明带上顶体反应精子的发生率显著更高。这些结果表明在HZA条件下人类配子功能具有高度的物种特异性,为该生物测定法在不孕症和避孕测试中的应用提供了进一步支持。