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Application of IVF to reproduction of a wild-derived strain MSM/Ms

机译：IVF在野生型MSM / Ms繁殖中的应用

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Reproduction technologies such as in vitro fertilization (IVF) and sperm freezing are powerful tools for not only reproduction research but also for a wide range of mouse genetics. Although reproduction technologies are well established for classical laboratory mouse strains, those for strains derived from wild mice have remained to be developed. An inbred strain MSM/Ms was established from Japanese wild mice, Mus musculus molossinus. Whole genome shot gun sequencing has been done with this strain, and a SNPs database of MSM/Ms is now open to public. We have established consomic strains carrying each MSM/Ms chromosome on the C57BL/6J background. Thus, MSM/Ms is becoming one of most useful mouse strain for genetic analysis. In this study, we examined super ovulation rate and IVF rate with MSM/Ms in order to facilitate reproduction of this useful strain. We administered 2.5IU of PMSG for MSM females intraperitoneally, and followed by administration of the same amount of hCG after 48 hours of the PMSG Then, oocytes were collected from the oviduct after 17 hours of the hCG administration. Number of the oocytes in ampulla of the oviduct was counted under the microscope for each experiment, followed by in vitro fertilization. Oocytes were inseminated with precultured spermatozoa that were collected from cauda epididymis of MSM/Ms males. When we administered PMSG and hCG for 4, 5, 6, 7, 8 weeks old and 9 to 27 weeks old females, average numbers of the ovulated oocytes obtained from the different aged females were 9.4, 10.0, 8.0, 8.0, 3.3, 4.4 respectively. Success rate of IVF for MSM/Ms was between 40% and 74% in average. These results indicated that super ovulation is effective for 4 to 8 weeks old MSM/Ms females, and that IVF is applicative to MSM/Ms.
机译：诸如体外受精（IVF）和精子冷冻之类的生殖技术不仅是生殖研究的重要工具，而且还是广泛的小鼠遗传学研究的有力工具。尽管对于经典的实验室小鼠品系而言，复制技术已经建立了很好的技术，但对于野生小鼠衍生品的复制技术仍有待开发。从日本野生小鼠小家鼠Mus musculus molossinus建立了近交系MSM / Ms。已经对该菌株进行了全基因组shot弹枪测序，并且MSM / Ms的SNPs数据库现已向公众开放。我们已经建立了携带C57BL / 6J背景上每个MSM / MS染色体的纯系菌株。因此，MSM / Ms成为遗传分析中最有用的小鼠品系之一。在这项研究中，我们检查了MSM / Ms的超排卵率和IVF率，以促进这种有用菌株的繁殖。我们腹膜内给予MSM女性2.5IU PMSG，然后在PMSG 48小时后给予等量的hCG。然后，在hCG给予17小时后从输卵管中收集卵母细胞。对于每个实验，在显微镜下计数输卵管壶腹中的卵母细胞数，然后进行体外受精。卵母细胞用从MSM / Ms雄性马尾附睾收集的预培养精子进行授精。当我们分别对4、5、6、7、8周龄的雌性和9至27周龄的雌性给予PMSG和hCG时，从不同年龄的雌性获得的排卵卵母细胞的平均数量为9.4、10.0、8.0、8.0、3.3、4.4分别。 MSM / Ms的IVF成功率平均在40％至74％之间。这些结果表明，超级排卵对4至8周龄的MSM / Ms女性有效，并且IVF适用于MSM / Ms。

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