Tateno Hiroyuki
Department of Biological Sciences Asahikawa Medical College 078-8510 Asahikawa Japan.
Reprod Med Biol. 2009 May 14;8(3):89-95. doi: 10.1007/s12522-009-0017-x. eCollection 2009 Sep.
Incidence of structural chromosome aberrations in mouse one-cell embryos produced by intracytoplasmic sperm injection (ICSI) with mature epididymal spermatozoa were influenced by sperm incubation medium and time. When spermatozoa were incubated in bicarbonate-buffered TYH for ≤0.5 h, the embryo aberration rates were significantly higher than in vitro fertilization (IVF) embryos. However, after the incubation of spermatozoa in the same medium for ≥2 h, the aberration rates were close to the IVF embryo level. When spermatozoa were incubated in bicarbonate-buffered mCZB, hepes-buffered H-TYH and H-mCZB, and phosphate-buffered PB1, the increased incidences of aberrations were observed at any incubation time. In the case of sperm incubation in H-TYH, H-mCZB and PB1, the aberration rates increased in a time-dependent manner. Chromosome aberrations generated by ICSI were transmissible to offspring. On the other hand, the aberration rate in embryos derived from testicular spermatozoa was independent of the medium type and incubation time. Thus, the incubation media appears to have no effect on sperm chromatin. TYH can effectively induce capacitation and acrosome reaction, while H-TYH, H-mCZB and PB1 never induce these spermatozoal events. It is probable that the cholesterol-rich plasma membrane and intact acrosome injected into the ooplasm affect sperm chromatin remodeling, thus resulting in the generation of chromosome damage in ICSI embryos.
用成熟附睾精子经胞浆内精子注射(ICSI)产生的小鼠单细胞胚胎中,结构染色体畸变的发生率受精子孵育培养基和时间的影响。当精子在碳酸氢盐缓冲的TYH中孵育≤0.5小时时,胚胎畸变率显著高于体外受精(IVF)胚胎。然而,当精子在相同培养基中孵育≥2小时后,畸变率接近IVF胚胎水平。当精子在碳酸氢盐缓冲的mCZB、HEPES缓冲的H-TYH和H-mCZB以及磷酸盐缓冲的PB1中孵育时,在任何孵育时间都观察到畸变发生率增加。在精子在H-TYH、H-mCZB和PB1中孵育的情况下,畸变率呈时间依赖性增加。ICSI产生的染色体畸变可传递给后代。另一方面,源自睾丸精子的胚胎中的畸变率与培养基类型和孵育时间无关。因此,孵育培养基似乎对精子染色质没有影响。TYH能有效诱导获能和顶体反应,而H-TYH、H-mCZB和PB1从未诱导这些精子事件。很可能注入卵母细胞质中的富含胆固醇的质膜和完整顶体影响精子染色质重塑,从而导致ICSI胚胎中染色体损伤的产生。