Live Births from Domestic Dog (Canis familiaris) Embryos Produced by In Vitro Fertilization.
Live Births from Domestic Dog (Canis familiaris) Embryos Produced by In Vitro Fertilization.
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DOI：
10.1371/journal.pone.0143930
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Travis AJ
中科院分区：
文献类型：
--
作者：
Nagashima JB;Sylvester SR;Nelson JL;Cheong SH;Mukai C;Lambo C;Flanders JA;Meyers-Wallen VN;Songsasen N;Travis AJ
几十年来，由于其独特的生殖生理学，辅助生殖技术（ART）在狗中的发展一直阻碍着进步。鉴于ART在保护濒危犬科动物物种或通过基因编辑技术根除遗传性疾病方面可以发挥关键作用，这种缺乏进展是值得注意的-这种方法也将推动狗作为生物医学模型。超过350种狗的遗传性疾病/特征与人类的情况同源，几乎是任何其他物种的两倍。在这里，我们报告了第一个活产的体外受精胚胎的狗。更重要的是，这些胚胎也被冷冻保存。两种配子处理方式的改变促成了这一进展。以前用于精子获能的培养基不包括镁，因为它延迟了自发的顶体胞吐。我们发现，镁显着增强精子的超活化和能力，进行生理诱导的顶体胞吐，两个功能的受精卵。与其他哺乳动物不同，狗排卵初级卵母细胞，在促黄体激素（LH）激增后的第4-5天达到中期II。我们发现只有在第6天卵母细胞才能持续受精。第6天卵母细胞与在补充镁的培养基中获能的精子进行体外受精，胚胎发育率很高（78.8%，n = 146）。19个冷冻保存的体外受精（IVF）胚胎的输卵管内移植产生了7只健康的小狗。体外受精的发展使现代遗传方法能够更有效地应用于狗，并用于配子拯救以保护濒危犬科动物。
Development of assisted reproductive technologies (ART) in the dog has resisted progress for decades, due to their unique reproductive physiology. This lack of progress is remarkable given the critical role ART could play in conserving endangered canid species or eradicating heritable disease through gene-editing technologies—an approach that would also advance the dog as a biomedical model. Over 350 heritable disorders/traits in dogs are homologous with human conditions, almost twice the number of any other species. Here we report the first live births from in vitro fertilized embryos in the dog. Adding to the practical significance, these embryos had also been cryopreserved. Changes in handling of both gametes enabled this progress. The medium previously used to capacitate sperm excluded magnesium because it delayed spontaneous acrosome exocytosis. We found that magnesium significantly enhanced sperm hyperactivation and ability to undergo physiologically-induced acrosome exocytosis, two functions essential to fertilize an egg. Unlike other mammals, dogs ovulate a primary oocyte, which reaches metaphase II on Days 4–5 after the luteinizing hormone (LH) surge. We found that only on Day 6 are oocytes consistently able to be fertilized. In vitro fertilization of Day 6 oocytes with sperm capacitated in medium supplemented with magnesium resulted in high rates of embryo development (78.8%, n = 146). Intra-oviductal transfer of nineteen cryopreserved, in vitro fertilization (IVF)-derived embryos resulted in seven live, healthy puppies. Development of IVF enables modern genetic approaches to be applied more efficiently in dogs, and for gamete rescue to conserve endangered canid species.