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利用改良的辅助生殖技术高效繁殖日本野生型小家鼠(Mus musculus molossinus)后代。

Efficient production of offspring from Japanese wild-derived strains of mice (Mus musculus molossinus) by improved assisted reproductive technologies.

机构信息

RIKEN BioResource Center, Tsukuba, Japan.

出版信息

Biol Reprod. 2012 May 31;86(5):167, 1-7. doi: 10.1095/biolreprod.111.098491. Print 2012 May.

Abstract

Because the genetic diversity of the laboratory mouse (Mus musculus) is very limited, wild-derived strains from this genus could provide invaluable experimental models for studies of mouse genetics and epigenetics such as quantitative trait locus analysis. However, such strains generally show poor reproductive performance under conventional husbandry conditions, so their use for large-scale analyses has been limited. This study was undertaken to devise assisted reproductive technologies (ARTs) for the efficient production of offspring in two wild-derived strains, MSM/Ms and JF1/Ms (Mus musculus molossinus). First, as females of these strains are poor responders to equine chorionic gonadotropin (eCG) stimulation, we examined the efficiency of superovulation by injecting anti-inhibin serum followed by human chorionic gonadotropin (hCG). Approximately four to six times more oocytes were ovulated than with eCG-hCG treatment in both strains, reaching ∼25-30 oocytes per female. Consequently, the procedures for in vitro fertilization using these superovulated oocytes and cryopreservation of embryos and spermatozoa could be optimized for both of the wild-derived strains. However, MSM/Ms embryos but not JF1/Ms embryos failed to develop to term after embryo transfer because of intrauterine death at mid to late gestation. We were able to overcome this obstacle by cotransfer of these embryos with those from laboratory strains combined with treatment of recipient females with an immunosuppressant (cyclosporin A). Thus, a series of ARTs essential for efficient production and preservation of the wild-derived strains were successfully devised. These technologies will facilitate systematic studies of mouse genetics and epigenetics using a wider range of genetic diversity than currently available in the genus Mus.

摘要

由于实验小鼠(Mus musculus)的遗传多样性非常有限,因此该属的野生衍生株系可为小鼠遗传学和表观遗传学的研究提供非常有价值的实验模型,例如数量性状基因座分析。然而,在常规饲养条件下,这些品系通常表现出较差的繁殖性能,因此它们的大规模应用受到限制。本研究旨在设计辅助生殖技术(ARTs),以有效生产两个野生衍生株系,MSM/Ms 和 JF1/Ms(Mus musculus molossinus)的后代。首先,由于这些株系的雌性对马绒毛膜促性腺激素(eCG)刺激的反应不佳,我们通过注射抗抑制素血清然后用人绒毛膜促性腺激素(hCG)检查了超数排卵的效率。与 eCG-hCG 处理相比,两种株系中都有大约四到六倍的卵母细胞排卵,达到每个雌性约 25-30 个卵母细胞。因此,可以针对两种野生衍生株系优化使用这些超排卵卵母细胞进行体外受精的程序以及胚胎和精子的冷冻保存。然而,MSM/Ms 胚胎而不是 JF1/Ms 胚胎在胚胎移植后未能发育至足月,因为在妊娠中期至晚期发生宫内死亡。我们能够通过将这些胚胎与来自实验室株系的胚胎一起转移并结合用免疫抑制剂(环孢素 A)处理受体雌性来克服这一障碍。因此,成功设计了一系列对于高效生产和保存野生衍生株系至关重要的 ARTs。这些技术将促进使用比目前 Mus 属中可用的遗传多样性更广泛的范围进行小鼠遗传学和表观遗传学的系统研究。

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