Shimizu Yasufumi, Minaguchi Reiko, Ishikawa Tomonori, Harada Tatsuya, Yoshiki Naoyuki, Kubota Toshiro
Department of Comprehensive Reproductive Medicine, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan.
Reprod Med Biol. 2008 Aug 3;7(3):143-149. doi: 10.1111/j.1447-0578.2008.00211.x. eCollection 2008 Sep.
The increase in the concentration of cytosolic-free calcium ([Ca]i) induced by follicular fluid or progesterone has been reported to promote an acrosome reaction and alternation in several motion parameters in human sperm (hyperactivation). We previously reported that populations of sperm in cell suspension obtained from infertile men with abnormal morphology exhibited lower mean peak progesterone-evoked [Ca]i compared with morphologically normal sperm using cell-suspension methods. In the present study, the change in [Ca]i in individual normally and abnormally shaped spermatozoa was compared. The change in [Ca]i induced by human follicular fluid in individual spermatozoa with normal and abnormal morphology was compared using the fluorescent calcium-sensitive dye fluo-3/AM. The spatial distribution of the increase in [Ca]i in single sperm was also investigated. The [Ca]i of normally shaped spermatozoa increased rapidly after the administration of human follicular fluid. The response reached a peak within 2-3 s and then slowly declined to a plateau phase. The baseline and peak fluorescence in spermatozoa with abnormal morphology was lower when compared with normal spermatozoa. The follicular-fluid-induced increase in [Ca]i (expressed as a percentage increase in [Ca]i over basal) in morphologically abnormal sperm was 39.2 ± 5.3% ( = 107, mean ± standard error), which was smaller than that of morphologically normal sperm (61.6 ± 5.7%, = 100, < 0.005) from seven healthy donors. The follicular-fluid-induced [Ca]i increases observed in sperm with morphologically abnormal mid-pieces (20.9 ± 4.3%, = 12, < 0.05) or tails (40.7 ± 6.0%, = 92, < 0.05) were lower than those of morphologically normal spermatozoa (61.6 ± 5.3%, = 101). The follicular-fluid-induced [Ca]i increase of morphologically normal spermatozoa from infertile couples (35.1 ± 6.3%, = 25, < 0.05) was also found to be lower than that of morphologically normal spermatozoa from healthy donors. The present study shows that spermatozoa with abnormal morphology in healthy donors have disorders of signal transduction, as do normally shaped sperm in men from infertile couples. (Reprod Med Biol 2008; : 143-149).
据报道，卵泡液或孕酮诱导的胞质游离钙浓度（[Ca]i）升高可促进人类精子的顶体反应和几种运动参数的改变（超活化）。我们之前报道，使用细胞悬液法，与形态正常的精子相比，从不育男性中获得的形态异常的细胞悬液中的精子群体，其孕酮诱发的平均[Ca]i峰值较低。在本研究中，比较了正常和异常形状的单个精子中[Ca]i的变化。使用荧光钙敏染料fluo-3/AM比较了人类卵泡液在形态正常和异常的单个精子中诱导的[Ca]i变化。还研究了单个精子中[Ca]i升高的空间分布。给予人类卵泡液后，形态正常的精子[Ca]i迅速升高。反应在2-3秒内达到峰值，然后缓慢下降至平台期。与正常精子相比，形态异常的精子的基线和峰值荧光较低。形态异常精子中卵泡液诱导的[Ca]i升高（以[Ca]i相对于基础值的百分比增加表示）为39.2±5.3%（n = 107，平均值±标准误），小于来自7名健康供体的形态正常精子（61.6±5.7%，n = 100，P < 0.005）。在形态异常的中段（20.9±4.3%，n = 12，P < 0.05）或尾部（40.7±6.0%，n = 92，P < 0.05）的精子中观察到的卵泡液诱导的[Ca]i升高低于形态正常的精子（61.6±5.3%，n = 101）。不育夫妇中形态正常的精子的卵泡液诱导的[Ca]i升高（35.1±6.3%，n = 25，P < 0.05）也低于健康供体中形态正常的精子。本研究表明，健康供体中形态异常的精子以及不育男性中形态正常的精子均存在信号转导障碍。（《生殖医学与生物学》2008年；：143 - 149）
