Wu H, He C L, Fissore R A
Program in Molecular and Cellular Biology, University of Massachusetts, Amherst 01003, USA.
Mol Reprod Dev. 1998 Jan;49(1):37-47. doi: 10.1002/(SICI)1098-2795(199801)49:1<37::AID-MRD5>3.0.CO;2-P.
Injection of sperm preparations into mammalian oocytes and eggs has been shown to elicit persistent [Ca2+]i oscillations that closely resemble fertilization-associated Ca2+ release. However, the ability of these sperm fractions to initiate egg activation has not been clearly demonstrated. In the present experiments, mouse eggs injected with a porcine sperm preparation were evaluated for early and late events of activation. Events monitored included, among early events, the generation of [Ca2+]i oscillations and cortical granule exocytosis and, among late events, the decrease in histone H1 and myelin basic protein kinase activities, polar body extrusion, pronuclear formation, and cleavage to the two-cell stage. Injection of sperm fractions consistently evoked [Ca2+]i oscillations that, in turn, initiated all events of activation. Uninjected control eggs or eggs injected with buffer or heat-treated sperm fractions failed to show Ca2+ responses or activation. In addition, injection of sperm fractions into recently ovulated eggs (experiments were concluded within 15 hr after human chorionic gonadotropin administration) induced high rates of activation, while similarly aged eggs exposed to 7% ethanol for 5 min, a known parthenogenetic treatment, failed to activate. Together these results indicate that injection of sperm fractions elicits [Ca2+]i oscillations that are capable of initiating normal egg activation. These results support the hypothesis that a sperm component participates in the generation of fertilization-associated [Ca2+]i oscillations.
将精子制剂注入哺乳动物的卵母细胞和卵子已被证明会引发持续的[Ca2+]i振荡,这种振荡与受精相关的Ca2+释放非常相似。然而,这些精子组分启动卵子激活的能力尚未得到明确证实。在本实验中,对注射了猪精子制剂的小鼠卵子进行了激活早期和晚期事件的评估。监测的事件包括,早期事件中有[Ca2+]i振荡的产生和皮质颗粒胞吐作用,晚期事件中有组蛋白H1和髓鞘碱性蛋白激酶活性的降低、极体排出、原核形成以及分裂至二细胞期。注射精子组分始终会引发[Ca2+]i振荡,进而启动所有激活事件。未注射的对照卵子或注射了缓冲液或热处理精子组分的卵子未表现出Ca2+反应或激活。此外,将精子组分注入刚排卵的卵子(在注射人绒毛膜促性腺激素后15小时内完成实验)会诱导高激活率,而同样年龄的卵子暴露于7%乙醇5分钟(一种已知的孤雌生殖处理方法)则未能激活。这些结果共同表明,注射精子组分引发的[Ca2+]i振荡能够启动正常的卵子激活。这些结果支持了一种假设,即精子的一种成分参与了受精相关的[Ca2+]i振荡的产生。