Effect of protein kinase C stimulators on zona pellucida binding and the acrosome reaction of macaque sperm.
Effect of protein kinase C stimulators on zona pellucida binding and the acrosome reaction of macaque sperm.
复制标题
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。
DOI：
10.1095/biolreprod52.6.1418
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发表时间：
1995
影响因子：
3.6
通讯作者：
VandeVoort,CA
中科院分区：
文献类型：
--
作者：
Tollner,TL;Overstreet,JW;VandeVoort,CA
猕猴精子需要用二丁酰cAMP(DBcAMP)和咖啡因(称为激活剂；ACT)处理才能在体外完全获能。以前的研究表明，ACT治疗还增加了精子与猕猴透明带的结合，并增强了结合精子的顶体反应的诱导。本研究探讨了刺激蛋白激酶C(PKC)的佛波酯(PMA)和1，2-二辛酰甘油(DOG)是否也能增加精子与透明带的结合和/或增强结合精子顶体反应的诱导。食蟹猴精子经60%Percoll离心后，用改良的Biggers，Whitten and Whittingham(BWW)培养液洗涤，在37℃，5%CO2条件下孵育2.5h，最后15~30min向精子中加入ACT、PMA或DOG。精子孵育后立即评估精子的活力和顶体状态。猕猴卵母细胞与刺激的精子悬液在BWW培养液中共孵育30秒(脉冲)。取一半的卵母细胞放入无精子培养液中继续孵育1h(Chase)。当在脉冲期后进行评估时，与未治疗的对照组相比，ACT治疗后精子-小带结合显著增加。狗而不是PMA处理对精子结合PMA有类似的影响，与对照组相比，狗和ACT处理增加了30秒脉冲后结合到透明带的精子顶体反应的百分比。PMA和DOG的非活性类似物对精子-透明带结合或结合精子顶体反应(%AR)的百分比没有影响。
Macaque sperm require treatment with dibutyryl cAMP (dbcAMP) and caffeine (termed activators; ACT) to fully capacitate in vitro. Previous studies have shown that treatment with ACT also increases sperm binding to the macaque zona pellucida and enhances the induction of acrosome reactions of the bound sperm. This study investigated whether phorbol 12-myristate 13-acetate (PMA) and 1, 2-dioctanoyl-sn-glycerol (DOG), which stimulate protein kinase C (PKC), can also increase sperm binding to the zona pellucida and/or enhance induction of the acrosome reaction of bound sperm. Cynomolgus macaque sperm were centrifuged through 60% Percoll and then were washed with modified Biggers, Whitten and Whittingham (BWW) medium and incubated for 2.5 h at 37 degrees C with 5% CO2, ACT, PMA, or DOG was added to sperm during the last 15-30 min of incubation. Sperm were evaluated immediately after incubation for motility and acrosomal status. Macaque oocytes were coincubated with stimulated sperm suspensions in BWW medium for 30 sec (pulse). Half of the oocytes were removed to sperm-free medium and further incubated for 1 h (chase). When assessed after the pulse period, sperm-zona binding increased significantly after ACT treatment compared to that in untreated controls. DOG but not PMA treatment had a similar effect on sperm binding PMA, DOG, and ACT treatment increased the percentage of acrosome reactions in sperm bound to the zona following the 30-sec pulse compared to the percentage in controls. Inactive analogs of PMA and DOG had no effect on sperm-zona binding or the percentage of acrosome reactions (% AR) of bound sperm.(ABSTRACT TRUNCATED AT 250 WORDS)