Harada Yoshihisa, Kinutani Masayuki, Horiuchi Toshitaka
Graduate School of Comprehensive Scientific Research Prefectural University of Hiroshima Hiroshima Japan.
Kinutani Women's Clinic Hiroshima Japan.
Reprod Med Biol. 2020 Feb 18;19(2):171-177. doi: 10.1002/rmb2.12319. eCollection 2020 Apr.
To investigate the first-division kinetics and in vitro development of embryos produced by injecting sonicated sperm heads with high or low chromosomal integrity into oocytes.
Mouse spermatozoa were frozen after separating the sperm heads from the tails by sonication in an EGTA solution (EGTA group) or M2 medium (M2 group). The chromosomal integrity of sonicated mouse spermatozoa was analyzed by injecting the sperm heads into fresh mouse oocytes. The developmental potential of spermatozoa was examined by injecting the sperm heads into vitrified-warming mouse oocytes. We used a time-lapse monitoring system to compare the first-division kinetics.
Chromosomal integrity was preserved significantly more frequently in the EGTA group (90.6%) than in the M2 group (32.7%). Blastocysts developed significantly more often in the EGTA group (80.8%) than in the M2 group (39.6%). In the M2 group, with frequent chromosome aberrations, the time between the sperm injection and first cleavage was delayed (18.4 hours), compared to the EGTA group (16.5 hours). All results of the EGTA group were similar to that of fresh epididymal spermatozoa.
The EGTA solution for sonication maintained the integrity of sperm chromosomes. Our results revealed a relationship between sperm chromosome integrity and first-division kinetics.
研究将具有高或低染色体完整性的超声处理精子头注射到卵母细胞中所产生胚胎的第一次分裂动力学和体外发育情况。
通过在EGTA溶液(EGTA组)或M2培养基(M2组)中超声处理将小鼠精子的头部与尾部分离后,将小鼠精子冷冻。通过将精子头注射到新鲜小鼠卵母细胞中,分析超声处理小鼠精子的染色体完整性。通过将精子头注射到玻璃化复温的小鼠卵母细胞中,检测精子的发育潜能。我们使用延时监测系统比较第一次分裂动力学。
EGTA组(90.6%)的染色体完整性保存频率显著高于M2组(32.7%)。EGTA组(80.8%)发育成囊胚的频率显著高于M2组(39.6%)。在M2组中,由于频繁出现染色体畸变,与EGTA组(16.5小时)相比,精子注射与第一次卵裂之间的时间延迟(18.4小时)。EGTA组的所有结果与新鲜附睾精子的结果相似。
用于超声处理的EGTA溶液保持了精子染色体的完整性。我们的结果揭示了精子染色体完整性与第一次分裂动力学之间的关系。