Peromyscus Genetic Stock Center, Department of Biological Sciences, University of South Carolina, Columbia, South Carolina, USA.
Theriogenology. 2012 Jan 15;77(2):311-9. doi: 10.1016/j.theriogenology.2011.07.044. Epub 2011 Sep 14.
Although laboratory-reared species of the genus Peromyscus-including deer mice-are used as model animals in a wide range of research, routine manipulation of Peromyscus embryogenesis and reproduction has been lagging. The objective of the present study was to optimize conditions for oocyte and/or embryo retrieval and for in vitro culturing. On average, 6.4 oocytes per mouse were recovered when two doses of 15 IU of pregnant mare serum gonadotropin (PMSG) were given 24 h apart, followed by 15 IU of hCG 48 h later. Following this hormone priming, females mated overnight with a fertile male yielded an average of 9.1 two-cell stage embryos. Although two-cell stage embryos developed to 8-cell stage in Potassium Simplex Optimized Medium (KSOM; Millipore-Chemicon, Billerica, MA, USA) in vitro, but not further, embryos recovered at the 8- to 16-cell stages developed into fully expanded blastocysts when cultured in M16 media in vitro. These blastocysts had full potential to develop into late stage fetuses and possibly into live pups. As a result of the present work, all stages of Peromyscus preimplantation development are now obtainable in numbers sufficient for molecular or other analyses. These advances provide the opportunity for routine studies involving embryo transfer (e.g., chimeras, transgenics), and preservation of genetic lines by cryopreservation.
虽然实验室饲养的 Peromyscus 属物种——包括鹿鼠——被广泛用于各种研究的模型动物,但 Peromyscus 胚胎发生和繁殖的常规操作一直滞后。本研究的目的是优化卵母细胞和/或胚胎回收和体外培养的条件。平均每只小鼠可回收 6.4 个卵母细胞,方法是在 24 小时内给予两剂 15 IU 孕马血清促性腺激素(PMSG),然后在 48 小时后给予 15 IU hCG。在这种激素诱导后,与一只可育的雄性进行一夜交配,可产生平均 9.1 个两细胞期胚胎。尽管两细胞期胚胎在 Potassium Simplex Optimized Medium(KSOM;Millipore-Chemicon,Billerica,MA,USA)中可体外发育至 8 细胞期,但不能进一步发育,在 M16 培养基中体外培养的 8-16 细胞期胚胎可发育成完全扩张的囊胚。这些囊胚具有完全发育成晚期胎儿并可能发育成活幼崽的潜力。由于本工作的结果,现在可以获得足够数量的 Peromyscus 植入前发育的所有阶段,用于分子或其他分析。这些进展为胚胎移植(例如嵌合体、转基因)和通过冷冻保存保存遗传系提供了机会。