Yoshimatsu N, Yanagimachi R, Lopata A
Department of Anatomy and Reproductive Biology, University of Hawaii School of Medicine, Honolulu 96822.
Gamete Res. 1988 Oct;21(2):115-26. doi: 10.1002/mrd.1120210202.
Zonae pellucidae of unfertilized hamster and human eggs remain penetrable by homologous spermatozoa when they are stored in highly concentrated salt solutions [Yanagimachi et al., 1979]. Neutralization of salt solutions with the appropriate pH buffers is recommended for long-term storage of the eggs (zonae), because zonae may gradually swell or even dissolve in acidic or alkaline solutions. At least for the hamster, 2 M ammonium sulfate solution containing 40 mM Hepes and 0.5% dextran (pH 7.0) seems to serve the purpose satisfactorily. Thorough rinsing of salt-stored zonae prior to insemination is important, but inclusion of protease inhibitors in the rinsing medium does not appear to be essential. A dialysis method is recommended when a large number of salt-stored eggs must be rinsed at one time. Salt-stored hamster zonae, like fresh (control) zonae, permit attachment and penetration of hamster spermatozoa, but not human spermatozoa. Salt-stored human zonae, on the other hand, allow penetration by not only human spermatozoa, but also hamster spermatozoa.
未受精的仓鼠和人类卵子的透明带保存在高浓度盐溶液中时,仍可被同种精子穿透[柳町等,1979年]。建议用适当的pH缓冲液中和盐溶液以长期保存卵子(透明带),因为透明带在酸性或碱性溶液中可能会逐渐肿胀甚至溶解。至少对于仓鼠来说,含有40 mM羟乙基哌嗪乙磺酸(HEPES)和0.5%葡聚糖(pH 7.0)的2 M硫酸铵溶液似乎能很好地满足这一目的。授精前对用盐保存的透明带进行彻底冲洗很重要,但冲洗介质中加入蛋白酶抑制剂似乎并非必需。当必须一次性冲洗大量用盐保存的卵子时,建议采用透析法。用盐保存的仓鼠透明带与新鲜(对照)透明带一样,允许仓鼠精子附着和穿透,但不允许人类精子穿透。另一方面,用盐保存的人类透明带不仅允许人类精子穿透,也允许仓鼠精子穿透。