Production of androgenetic diploid loach by cold-shock of eggs fertilized with diploid sperm
Production of androgenetic diploid loach by cold-shock of eggs fertilized with diploid sperm
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DOI：
10.1016/j.theriogenology.2013.03.014
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发表时间：
2013-07-15
期刊：
影响因子：
2.8
通讯作者：
Arai, Katsutoshi
中科院分区：
文献类型：
--
作者：
Hou, Jilun;Fujimoto, Takafumi;Arai, Katsutoshi
泥鳅(Misgurnus Anguillicaudatus)在无卵照射的情况下产生二倍体雄核。野生型二倍体雌性卵子与新四倍体雄性的二倍体精子受精，受精后立即在3℃(范围为+/-0.5℃)进行30分钟的冷休克处理，以消除雌性核。孵化后，用流式细胞仪对孵化后的幼虫进行倍性分析，发现可能是二倍体雄鱼，也有其他倍体的幼虫。对5个独立的微卫星DNA标记进行了基因分型，以确认所获得的二倍体幼虫的全雄性遗传。冷休克组二倍体雄核发育幼虫的平均+/-SD率为12.29+/-3.25%，紫外线照射组为22.23+/-13.42%(P>；0.05)。正常对照组未检测到二倍体雄核发育幼虫。据我们所知，这是第一个在没有卵子照射的情况下成功诱导鱼类二倍体雄核基因的报告。(C)2013 Elsevier Inc.保留所有权利。
Diploid androgenotes were produced without egg irradiation in the loach, Misgurnus anguillicaudatus. Eggs of wild-type diploid females were fertilized with diploid sperm of a neo-tetraploid male and then cold-shock treated at 3 degrees C (range, +/- 0.5 degrees C) for 30 minutes just after fertilization to eliminate the female nucleus. After hatching, ploidy status of the hatched larvae was analyzed by flow cytometry, which revealed putative diploid androgenotes as well as larvae possessing other ploidies. Five independent microsatellite DNA markers were genotyped to confirm all-male inheritance of the resultant diploid larvae. The mean +/- SD yield rate of diploid androgenetic larvae to total eggs used was 12.29 +/- 3.25% in the cold-shock group and 22.23 +/- 13.42% in the UV-irradiated group (P > 0.05). No diploid androgenetic larvae were detected in the intact control group. To our knowledge, this is the first report demonstrating successful induction of diploid androgenotes without egg irradiation in fish. (C) 2013 Elsevier Inc. All rights reserved.