Boediono A, Saha S, Sumantri C, Suzuki T
United Graduate School of Veterinary Sciences, Yamaguchi University, Japan.
Reprod Fertil Dev. 1995;7(5):1073-9. doi: 10.1071/rd9951073.
Mature bovine oocytes were activated with 7% ethanol followed by cytochalasin B or D treatment. Most oocytes extruded a second polar body and formed one pronucleus when treated with 7% ethanol alone [35/43 (81%)]. With ethanol followed by cytochalasin B or D, overall activation frequency was 70% (309/441), with activated oocytes containing two pronuclei. The cleavage rate was not significantly different between treatment with ethanol alone and ethanol followed by 5 micrograms mL-1 cytochalasin B, but it was significantly lower than in fertilized oocytes (P < 0.01). However, the blastocyst production rate was significantly different (P < 0.01) among the treatments. The incidence of parthenogenetic embryos with normal (diploid) complements and with chromosome anomalies (2N/4N) was 68% (17/25) and 32% (8/25) respectively, and this was not affected by cryopreservation treatment. The longitudinal diameter of aggregated-four embryos cultured in vitro was greater (P < 0.01) than aggregated-two or single embryos. One of the aggregated-four parthenogenetic embryos was further cultured in vitro and developed up to Day 27 after activation, with a diameter of 2980 microns. The aggregated-four parthenogenetic embryos were transferred to five recipients. The oestrus was prolonged in three recipients and they returned to oestrus on Day 57, 62 and 67 after the previous oestrus. These results indicate that aggregating parthenogenetic embryos can prolong their survival in vitro and in vivo.
成熟的牛卵母细胞先用7%乙醇激活，然后进行细胞松弛素B或D处理。单独用7%乙醇处理时，大多数卵母细胞排出第二极体并形成一个原核[35/43(81%)]。先用乙醇处理，再用细胞松弛素B或D处理，总体激活频率为70%(309/441)，激活的卵母细胞含有两个原核。单独用乙醇处理和先用乙醇再用5微克/毫升细胞松弛素B处理之间的卵裂率没有显著差异，但显著低于受精的卵母细胞(P<0.01)。然而，不同处理之间的囊胚形成率有显著差异(P<0.01)。具有正常(二倍体)染色体组成和染色体异常(2N/4N)的孤雌生殖胚胎的发生率分别为68%(17/25)和32%(8/25)，且不受冷冻保存处理的影响。体外培养的四个胚胎聚集体的纵径大于两个胚胎聚集体或单个胚胎(P<0.01)。将一个四个胚胎聚集体的孤雌生殖胚胎进一步体外培养，激活后发育到第27天，直径为2980微米。将四个胚胎聚集体的孤雌生殖胚胎移植到五个受体中。三个受体的发情期延长，它们在前次发情后的第57、62和67天恢复发情。这些结果表明，聚集孤雌生殖胚胎可以延长它们在体外和体内的存活时间。
