Development of assisted reproductive technologies for Mus spretus
Development of assisted reproductive technologies for Mus spretus
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DOI：
10.1093/biolre/ioaa177
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发表时间：
2021-01-01
影响因子：
3.6
通讯作者：
Ogura, Atsuo
中科院分区：
文献类型：
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作者：
Hasegawa, Ayumi;Mochida, Keiji;Ogura, Atsuo
鼠属由许多具有高度遗传多样性的物种组成。然而，只有一种物种——小家鼠（Mus musculus）（实验室小鼠）在生物医学研究中很常见。其他鼠属物种无法获得辅助生殖技术（ART）可能是其在实验室中使用有限的主要原因。在这里，我们为 Mus spretus（阿尔及利亚小鼠）设计了 ART，这是一种常用的野生小鼠物种。我们发现，由于马绒毛膜促性腺激素或抗抑制素血清（AIS）（每只雌性5-8个卵母细胞）超排的效率低以及体外受精后的受精率低（IVF；15.2％），体外生产M. spretus胚胎很困难。实验室小鼠相互体外受精显示，其主要原因是透明带硬化，而不是精子的受精能力。当雌性注射AIS，24小时后注射人绒毛膜促性腺激素和雌二醇，然后自然交配时，获得了最大数量的胚胎（每只雌性16个）。使用基于乙二醇的溶液，可以对这些体内衍生的 2 细胞胚胎进行玻璃化/加热，并具有高存活率 (94%)。重要的是，在种间胚胎移植到 (C57BL/6 x C3H) F1 雌性小鼠中后，超过 60% 的此类胚胎发育成健康的后代。因此，我们为小家鼠设计了实用的 ART，能够高效生产胚胎和动物，并在实验室安全保存其品系。此外，我们还证明了小鼠啮齿动物中种间胚胎移植是可能的。摘要使用新开发的辅助生殖技术方案，可以有效地生产、冷冻保存野生小鼠的胚胎，并将其移植到实验室雌性小鼠中。
The genus Mus consists of many species with high genetic diversity. However, only one species, Mus musculus (the laboratory mouse), is common in biomedical research. The unavailability of assisted reproductive technologies (ARTs) for other Mus species might be a major reason for their limited use in laboratories. Here, we devised ARTs for Mus spretus (the Algerian mouse), a commonly used wild-derived Mus species. We found that in vitro production of M. spretus embryos was difficult because of low efficacies of superovulation with equine chorionic gonadotropin or anti-inhibin serum (AIS) (5-8 oocytes per female) and a low fertilization rate following in vitro fertilization (IVF; 15.2%). The primary cause of this was the hardening of the zona pellucida but not the sperm's fertilizing ability, as revealed by reciprocal IVF with laboratory mice. The largest number of embryos (16 per female) were obtained when females were injected with AIS followed by human chorionic gonadotropin and estradiol injections 24 h later, and then by natural mating. These in vivo-derived 2-cell embryos could be vitrified/warmed with a high survival rate (94%) using an ethylene glycol-based solution. Importantly, more than 60% of such embryos developed into healthy offspring following interspecific embryo transfer into (C57BL/6 x C3H) F1 female mice. Thus, we have devised practical ARTs for Mus spretus mice, enabling efficient production of embryos and animals, with safe laboratory preservation of their strains. In addition, we have demonstrated that interspecific embryo transfer is possible in murine rodents.Summary sentenceEmbryos from wild-derived Mus spretus mice can be efficiently produced, cryopreserved, and transferred into laboratory mouse females using newly developed assisted reproductive technology protocols.