Preservation of ejaculated mouse spermatozoa from fertile C57BL/6 and infertile Hook1/Hook1 mice collected from the uteri of mated females
Preservation of ejaculated mouse spermatozoa from fertile C57BL/6 and infertile Hook1/Hook1 mice collected from the uteri of mated females
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DOI：
10.1095/biolreprod.106.059881
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发表时间：
2007-06-01
影响因子：
3.6
通讯作者：
Ward, Monika A.
中科院分区：
文献类型：
--
作者：
Yamauchi, Yasuhiro;Ward, Monika A.
常规用于保存小鼠精子的方法需要杀死雄性小鼠以从附睾中回收精子。在这里，我们从正常可育的 C57BL/6 和不育的 Hook1/Hook1（以前称为 azh/azh）突变雄性交配后的子宫中获得了多个射精精子样本，从而避免了雄性的终止。射出的精子通过常规冷冻保存或不进行冷冻保护的快速冷冻保存，并通过胞浆内单精子注射（ICSI）注射到卵母细胞中。当比较野生型小鼠与 Hook1/Hook1 小鼠以及测试小鼠与对照小鼠（新鲜和快速冷冻的附睾以及新鲜射精的精子）的两种保存方法时，存活、激活和发育成双细胞胚胎的卵母细胞的比例相似。将两细胞胚胎移植到假孕雌性的输卵管中，并在妊娠第 15 天检查胎儿发育情况。总共 39%-54% 的移植胚胎中植入了保存的射精精子。所有检查组和研究中的所有男性均获得活的正常胎儿（11%-17%）。在对照组中注意到更多的植入物（71%-82%）和胎儿（28%-31%）。用保存的射精精子产生的胚胎发育潜力较低可能是由于其获能状态所致；从子宫中取出的大部分精子均已获能。本研究对于小鼠新品系的维护和推广具有重要意义。该方法适用于所有类型的小鼠，包括患有男性不育综合症的小鼠。唯一的要求是感兴趣的雄性能够交配并且其射精含有精子。
Methods routinely used to preserve mouse spermatozoa require that the male be killed to recover spermatozoa from the epididymides. Here we obtained multiple samples of ejaculated spermatozoa from normal fertile C57BL/6 and infertile Hook1/Hook1 (formerly known as azh/azh) mutant males from uteri after mating, thus avoiding termination of the males. Ejaculated sperm were preserved by conventional cryopreservation or by rapid freezing without cryoprotection, and were injected into the oocytes by intracytoplasmic sperm injection (ICSI). The proportions of oocytes that survived, became activated, and developed into two-cell embryos were similar when comparing the two preservation methods in wildtype versus Hook1/Hook1 mice and tested mice versus controls (fresh and rapid-frozen epididymal and fresh ejaculated sperm). Two-cell embryos were transferred into the oviducts of pseudopregnant females, and fetal development was examined at Day 15 of gestation. A total of 39%-54% of transferred embryos produced with preserved ejaculated sperm implanted. Live, normal fetuses (11%-17%) were obtained in all examined groups and from all males included in the study. More implants (71%-82%) and fetuses (28%-31%) were noted in controls. Lower developmental potentials of embryos produced with preserved ejaculated sperm might be due to their capacitation status; the majority of sperm retrieved from the uterus were capacitated. This study bears significance for the maintenance and distribution of novel mouse strains. The method is applicable for all types of mice, including those with male infertility syndromes. The sole requirement is that the male of interest is able to copulate and its ejaculate contains spermatozoa.