Protein kinase C plays an important role in the human zona pellucida-induced acrosome reaction.
Protein kinase C plays an important role in the human zona pellucida-induced acrosome reaction.
复制标题
蛋白激酶C在人类透明带诱导的顶体反应中发挥重要作用。
作者：
D. Y. Liu;H. Baker
为探讨蛋白激酶在透明质酸（ZP）诱导的人顶体反应（AR）中的信号转导作用，研究了蛋白激酶（PK）激活剂二丁酰环磷酸腺苷（PKA）和环磷酸鸟苷（PKG）、佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA，PKC）及PKC抑制剂星形孢菌素（Staurosporine）对AR的影响。精子样本来自具有正常精子-ZP结合的正常精子男性。卵母细胞取自其他体外受精失败的患者。将通过上游技术选择的活动精子与2.5 - 20 μ M PMA、1 mM二丁酰cAMP或cGMP、3 mM戊酰茶碱或0.125 - 2.0 μ M星形孢菌素预孵育30分钟，然后与4个卵母细胞在补充有牛血清白蛋白的人输卵管液中孵育2小时。通过用小口径移液管反复抽吸卵母细胞，将与ZP结合的精子排出，并通过荧光素标记的豌豆凝集素确定AR的状态。精子与PMA孵育30 min和2 h后，用计算机辅助精子分析（CASA）评价精子活力和运动特征。二丁酰cAMP和cGMP类似物对ZP诱导的AR有轻微的促进作用（P <0.05），而戊乙茶碱对ZP诱导的AR无促进作用（P> 0.05）。与此相反，PMA刺激ZP诱导的AR在一个显着的剂量依赖性方式。只有最高浓度（15 - 20 μ M）的PMA显著降低运动百分比（P <0.001）。2.5 - 15 μ M的PMA显著刺激ZP诱导的AR而不降低运动性（P <0.001）。PKC抑制剂staurosporine（0.125 - 0.25 μ M）显著抑制ZP诱导的AR，但不影响运动（P <0.001）。来自33名正常精子男性的精子样本用于研究用15 μ M PMA增强的ZP诱导的AR。一个样本没有显示出对PMA刺激的反应。在14名低ZP诱导AR的男性中，一半对PMA有正常反应，另一半对PMA有低反应。总之，PKC的激活或抑制显着增加或减少人ZP诱导的AR表明，PKC在生理AR的信号转导途径中起着重要作用。
To investigate the involvement of protein kinases in signal transduction in the human zona pellucida (ZP)-induced acrosome reaction (AR), the effects of protein kinase (PK) activators, dibutyryl cAMP (PKA) and cGMP (PKG), phorbol 12-myristate 13-acetate (PMA, PKC), and the PKC inhibitor, staurosporine were studied. Sperm samples were obtained from normozoospermic men with normal sperm-ZP binding. Oocytes were obtained from other patients with failure of fertilization in vitro. Motile spermatozoa selected by a swim-up technique were pre-incubated with 2.5-20 microM PMA, 1 mM dibutyryl cAMP or cGMP, 3 mM pentoxifylline or 0.125-2.0 microM staurosporine for 30 min and then incubated with four oocytes for 2 h in human tubal fluid supplemented with bovine serum albumin. The spermatozoa bound to the ZP were dislodged by repeatedly aspirating the oocytes with a small bore pipette and the state of the AR was determined by fluorescein-labelled Pisum sativum agglutinin. Motility and movement characteristics were assessed by computer-assisted sperm analysis (CASA) after incubation of spermatozoa with PMA for 30 min and 2 h. The dibutyryl cAMP and cGMP analogues had a small positive effect (P < 0.05) but pentoxifylline had no effect on stimulating the ZP-induced AR (P > 0.05). In contrast, PMA stimulated ZP-induced AR in a marked dose-dependent manner. Only the highest concentrations (15-20 microM) of PMA significantly decreased percentage motility (P < 0.001). Doses of 2.5-15 microM of PMA significantly stimulated ZP-induced AR without decreasing motility (P < 0.001). The PKC inhibitor, staurosporine (0.125-0.25 microM) significantly inhibited ZP-induced AR without affecting motility (P < 0.001). Sperm samples from 33 normozoospermic men were used for studies of the ZP-induced AR augmented with 15 microM PMA. One sample did not show a response to PMA stimulation. Among the 14 men with low ZP-induced AR, half had normal responses to PMA and other half had low responses to PMA. In conclusion, activation or inhibition of PKC significantly increases or decreases human ZP-induced AR suggesting that PKC plays a important role in the signal transduction pathway for the physiological AR.
蛋白激酶 C 刺激剂对透明带结合和猕猴精子顶体反应的影响。
影响因子：
3.6
作者：
Tollner,TL;Overstreet,JW;VandeVoort,CA
通讯作者：
VandeVoort,CA
影响因子：
3.6
作者：
CROSS, NL;MORALES, P;HANSON, FW
通讯作者：
HANSON, FW