In vitro fertilization of in vitro matured minke whale (Balaenoptera acutorostrata) follicular oocytes
In vitro fertilization of in vitro matured minke whale (Balaenoptera acutorostrata) follicular oocytes
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DOI：
10.1111/j.1748-7692.1997.tb00647.x
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发表时间：
1997-07-01
影响因子：
2.3
通讯作者：
Ohsumi, S
中科院分区：
文献类型：
--
作者：
Fukui, Y;Mogoe, T;Ohsumi, S
对1995年11月至1996年3月在南极海域捕获的55头小须鲸（Balaenoptera acutorostrata）的卵泡卵母细胞进行了体外受精试验。在实验1中，研究了培养时间（96小时或120小时）对卵泡卵母细胞成熟的影响以及添加咖啡因（5 mM）和/或肝素（100 μ g/ml）对精子穿透和原核形成的影响。将从4只成熟雄性输精管中回收的精子稀释（5倍），并在-80 ℃下冷冻。解冻后和合并的精子用于体外授精。培养120 h的卵母细胞（34.2%/260）的第二中期比例高于培养96 h的卵母细胞（26.0%/262）（P < 0.05）。培养120 h的卵母细胞穿透率（55.1%）和原核形成率（40.4%）均显著高于培养96 h的卵母细胞穿透率（32.4%）和原核形成率（20.6%）（P < 0.05和P < 0.01）。添加咖啡因和肝素没有显示出显著的效果。在实验2中，卵泡卵母细胞成熟120小时，然后受精培养，以检查在两个培养系统（有和没有共培养的卵丘细胞）的后续发展。在448个受精卵母细胞中，观察到有（5.8%）和无（4.9%）共培养系统的卵裂胚胎（2-16个细胞）。54个未受精卵未观察到卵裂。这些结果表明，小须鲸体外成熟卵泡卵母细胞与冷冻保存的精子体外受精是可能的，产生卵裂胚胎。
In vitro fertilization of follicular oocytes harvested from ovaries and matured in vitro was attempted for 55 minke whales (Balaenoptera acutorostrata) captured for Japanese research purposes in the Antarctic Ocean during the period from November 1995 to March 1996. In Experiment 1, effects of culture duration (96 h or 120 h) on maturation of follicular oocytes and addition of caffeine (5 mM) and/or heparin (100 mu g/ml) on sperm penetration and pronuclear formation were investigated. Spermatozoa recovered from the vasa deferentia of four mature males were diluted (5-fold) and frozen at -80 degrees C. The post-thawed and pooled spermatozoa were used for in vitro insemination. A higher (P < 0.05) proportion of the oocytes cultured for 120 h (34.2% of 260) progressed beyond the second metaphase stage than of the oocytes cultured for 96 h (26.0% of 262). For the matured oocytes, higher rates of penetration (P < 0.05) and pronuclear formation (P < 0.01) were obtained in the oocytes cultured for 120 h (55.1% and 40.4%) than in those cultured for 96 h (32.4% and 20.6%). Addition of caffeine and heparin did not show a significant effect. In Experiment 2, follicular oocytes matured for 120 h and then inseminated were cultured to examine the subsequent development in two culture systems (with and without co-cultured cumulus cells). Of 448 inseminated oocytes, cleaved embryos (2-16 cells) were observed with (5.8%) and without (4.9%) co-cultured systems. No cleavage was observed in 54 ova without insemination. These results indicate that in vitro fertilization of minke whale in vitro matured follicular oocytes with cryopreserved spermatozoa is possible, yielding cleaved embryos.