Fisch J D, Behr B, Conti M
Division of Reproductive Endocrinology and Infertility, Department of Gynecology and Obstetrics, Stanford University School of Medicine, CA 94305, USA.
Hum Reprod. 1998 May;13(5):1248-54. doi: 10.1093/humrep/13.5.1248.
Inhibition of sperm phosphodiesterase (PDE) has been shown to increase cAMP concentrations and stimulate motility and the acrosome reaction. While several PDE genes exist in mammals, little is known about the physiological role of PDE forms expressed in human spermatozoa. Using type-selective inhibitors, we identified two of the PDE forms expressed in human spermatozoa and studied their involvement in sperm function. Selective inhibitors of calcium-calmodulin-regulated PDE1 (8-methoxy-isobutyl-methylxanthine) and cAMP-specific PDE4 (RS-25344, Rolipram) were used to study PDE forms in human sperm extracts. 8-MeIBMX and Rolipram/RS-25344 inhibited sperm PDE activity by 35-40 and 25-30% respectively. Subcellular fractionation of the sperm homogenate suggests these pharmacologically distinct forms may be located in separate cellular regions. To evaluate the functional significance of different PDE forms, the effect of type-specific PDE inhibition on sperm motility and the acrosome reaction was examined. PDE4 inhibitors enhanced sperm motility over controls without affecting the acrosome reaction, while PDE1 inhibitors selectively stimulated the acrosome reaction. These data indicate at least two distinct PDE types exist in human spermatozoa. Our findings also support the hypothesis that PDE subtypes affect sperm function by regulating separate pools of cAMP and may ultimately offer novel treatments to infertile couples with abnormal semen parameters.
抑制精子磷酸二酯酶(PDE)已被证明可提高环磷酸腺苷(cAMP)浓度,并刺激精子活力和顶体反应。虽然哺乳动物中存在几种PDE基因,但对于人类精子中表达的PDE形式的生理作用知之甚少。我们使用类型选择性抑制剂鉴定了人类精子中表达的两种PDE形式,并研究了它们在精子功能中的作用。使用钙调蛋白调节的PDE1(8-甲氧基异丁基甲基黄嘌呤)和cAMP特异性PDE4(RS-25344,咯利普兰)的选择性抑制剂来研究人类精子提取物中的PDE形式。8-甲氧基异丁基甲基黄嘌呤和咯利普兰/RS-25344分别抑制精子PDE活性35-40%和25-30%。精子匀浆的亚细胞分级分离表明,这些药理学上不同的形式可能位于不同的细胞区域。为了评估不同PDE形式的功能意义,研究了类型特异性PDE抑制对精子活力和顶体反应的影响。PDE4抑制剂增强了精子活力,超过了对照组,而不影响顶体反应,而PDE1抑制剂选择性地刺激了顶体反应。这些数据表明人类精子中至少存在两种不同类型的PDE。我们的研究结果还支持这样一种假设,即PDE亚型通过调节不同的cAMP池来影响精子功能,并最终可能为精液参数异常的不育夫妇提供新的治疗方法。