Yanagida K, Katayose H, Hirata S, Yazawa H, Hayashi S, Sato A
Department of Obstetrics and Gynecology, Fukushima Medical University, Fukushima, Japan and. Department of Obstetrics and Gynecology, Yamanashi Medical University, Yamanashi, Japan.
Hum Reprod. 2001 Jan;16(1):148-152. doi: 10.1093/humrep/16.1.148.
Sperm immobilization prior to intracytoplasmic sperm injection (ICSI) is thought to be necessary for efficient fertilization. A variety of methods of sperm immobilization (pipetting, squeezing and piezo application) are currently employed in ICSI. The effect of differences in immobilization method on the timing of initial Ca(2+) oscillations of oocytes in ICSI was investigated. Motile spermatozoa were immobilized in eosin Y solution using pipetting, squeezing and piezo application. Complete staining of the sperm head was achieved after 220.7, 42.2 and 5.0 s respectively. Oscillations after ICSI were measured fluorometrically for each method. The onset of Ca(2+) oscillations was observed at 4.8 to 80.4 min after ICSI. Ca(2+) oscillations developed earlier with the piezo method (14.4 +/- 6.4 min) than other methods (pipetting, 43.1 +/- 20.2 min, P < 0.01; squeezing, 18.4 +/- 3.8 min, P = NS). The piezo method produced the earliest staining of the sperm head and may have caused the most damage to the sperm membrane. A more rapid onset of Ca(2+) oscillations was also observed with the piezo method. The method of sperm immobilization may be important for the rapid release of sperm factors that initiate oocyte activation. This study also showed that Ca(2+) oscillations develop earlier in human oocytes treated by ICSI than indicated in previous reports.
胞浆内单精子注射（ICSI）前精子制动被认为对有效受精是必要的。目前ICSI中采用了多种精子制动方法（移液器吹吸、挤压和压电应用）。研究了制动方法的差异对ICSI中卵母细胞初始Ca(2+)振荡时间的影响。使用移液器吹吸、挤压和压电应用将活动精子制动于伊红Y溶液中。精子头部分别在220.7、42.2和5.0秒后完全染色。对每种方法通过荧光测定法测量ICSI后的振荡情况。在ICSI后4.8至80.4分钟观察到Ca(2+)振荡的开始。与其他方法相比，压电方法导致Ca(2+)振荡更早出现（14.4±6.4分钟）（移液器吹吸法为43.1±20.2分钟，P<0.01；挤压法为18.4±3.8分钟，P=无显著差异）。压电方法使精子头部染色最早，并且可能对精子膜造成的损伤最大。还观察到压电方法使Ca(2+)振荡更快开始。精子制动方法对于启动卵母细胞激活的精子因子的快速释放可能很重要。本研究还表明，通过ICSI处理的人类卵母细胞中Ca(2+)振荡比以前报道的出现得更早。
