Menzel Viviana A, Hinsch Elvira, Hägele Wolfgang, Hinsch Klaus-Dieter
Center of Dermatology and Andrology, Justus Liebig University Giessen, Giessen, Germany.
Asian J Androl. 2007 Sep;9(5):650-8. doi: 10.1111/j.1745-7262.2007.00240.x.
To investigate if the phytoestrogen, genistein, affects essential functions of cryopreserved bovine spermatozoa.
The effect of genistein upon motility was assessed by computer-assisted motion analysis. Hemizona assay was performed to detect the ability of spermatozoa binding to the zona pellucida. The inducibility of the acrosome reaction using progesterone and ZP3-6 peptide was analysed by fluorescein-conjugated Pisum sativum agglutinin (FITC-PSA)/Hoechst 33258 double staining. Capacitation after incubation with genistein was assessed by the chlortetracycline (CTC) assay. Immunoblots showed the pattern of protein tyrosine phosphorylation of cryopreserved bovine spermatozoa.
Immunodetection of tyrosine-phosphorylated proteins showed that genistein did not affect tyrosine phosphorylation in cryopreserved bovine spermatozoa. However, genistein significantly reduced the progesterone- and ZP3-6 peptide-mediated induction of the acrosome reaction and led to a dose-dependent inhibition of sperm-zona pellucida binding; while sperm motility and capacitation were not affected by this phytoestrogen, as indicated by computer-assisted sperm motion analysis and the CTC assay, respectively.
Our results suggest that in cryopreserved bovine spermatozoa, genistein affects a protein tyrosine phosphorylation-independent signal transduction pathway that is involved in sperm capacitation, the acrosome reaction and sperm-zona pellucida binding.
研究植物雌激素染料木黄酮是否会影响冷冻保存的牛精子的基本功能。
通过计算机辅助运动分析评估染料木黄酮对精子活力的影响。进行半透明带试验以检测精子与透明带结合的能力。使用荧光素偶联的豌豆凝集素（FITC-PSA）/Hoechst 33258双重染色分析用孕酮和ZP3-6肽诱导顶体反应的情况。通过金霉素（CTC）试验评估与染料木黄酮孵育后的获能情况。免疫印迹显示冷冻保存的牛精子的蛋白质酪氨酸磷酸化模式。
酪氨酸磷酸化蛋白的免疫检测表明，染料木黄酮不影响冷冻保存的牛精子中的酪氨酸磷酸化。然而，染料木黄酮显著降低了孕酮和ZP3-6肽介导的顶体反应诱导，并导致精子与透明带结合的剂量依赖性抑制；而计算机辅助精子运动分析和CTC试验分别表明，这种植物雌激素不影响精子活力和获能。
我们的结果表明，在冷冻保存的牛精子中，染料木黄酮影响一条独立于蛋白质酪氨酸磷酸化的信号转导途径，该途径参与精子获能、顶体反应和精子与透明带结合。
