Ogonuki Narumi, Inoue Hiroki, Matoba Shogo, Kurotaki Yoko K, Kassai Hidetoshi, Abe Yukiko, Sasaki Erika, Aiba Atsu, Ogura Atsuo
RIKEN BioResource Center, Tsukuba, Ibaraki, Japan.
Department of Marmoset Research, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan.
Mol Reprod Dev. 2018 May;85(5):376-386. doi: 10.1002/mrd.22971. Epub 2018 Mar 23.
The common marmoset (Callithrix jacchus) represents a promising nonhuman primate model for the study of human diseases because of its small size, ease of handling, and availability of gene-modified animals. Here, we aimed to devise reproductive technology for marmoset spermatid injection using immature males for a possible rapid generational turnover. Spermatids at each step could be identified easily by their morphology under differential interference microscopy: thus, early round spermatids had a round nucleus with a few nucleolus-like structures and abundant cytoplasm, as in other mammals. The spermatids acquired oocyte-activating capacity at the late round spermatid stage, as confirmed by the resumption of meiosis and Ca oscillations upon injection into mouse oocytes. The spermatids could be cryopreserved efficiently with a simple medium containing glycerol and CELL BANKER®. Late round or elongated spermatids first appeared at 10-12 months of age, 6-8 months before sexual maturation. Marmoset oocytes microinjected with frozen-thawed late round or elongated spermatids retrieved from a 12-month-old male marmoset developed to the 8-cell stage without the need for artificial oocyte activation stimulation. Thus, it might be possible to shorten the intergeneration time by spermatid injection, from 2 years (by natural mating) to 13-15 months including gestation.
普通狨猴(Callithrix jacchus)因其体型小、易于操作且有基因修饰动物可用,是一种很有前景的用于人类疾病研究的非人类灵长类动物模型。在此,我们旨在设计一种利用未成熟雄性狨猴进行精子细胞注射的生殖技术,以实现可能的快速代际更替。在微分干涉显微镜下,每个阶段的精子细胞都可以通过其形态轻松识别:因此,早期圆形精子细胞有一个圆形细胞核,带有一些核仁样结构和丰富的细胞质,这与其他哺乳动物一样。如将精子细胞注射到小鼠卵母细胞后减数分裂的恢复和钙离子振荡所证实的,精子细胞在圆形精子细胞后期获得了卵母细胞激活能力。精子细胞可以用含有甘油和CELL BANKER®的简单培养基高效冷冻保存。晚期圆形或伸长的精子细胞首次出现在10 - 12个月大时,比性成熟早6 - 8个月。用从一只12个月大的雄性狨猴获取的经冷冻解冻的晚期圆形或伸长精子细胞进行显微注射的狨猴卵母细胞,无需人工卵母细胞激活刺激即可发育到8细胞阶段。因此,通过精子细胞注射有可能将代际时间从2年(通过自然交配)缩短至包括妊娠期在内的13 - 15个月。