Mori C, Hashimoto H, Hoshino K
Department of Anatomy, Faculty of Medicine, Kyoto University, Japan.
Biol Reprod. 1988 Oct;39(3):737-42. doi: 10.1095/biolreprod39.3.737.
Fluorescence microscopic investigations of nuclear DNA stained with mithramycin or 4',6-diamidino-2-phenylindole (DAPI) were undertaken in oocytes and zygotes during in vitro fertilization (IVF) and subsequent early embryo development in mice. Mithramycin and DAPI bind stoichiometrically with G-C and A-T pairs of DNA, respectively. In the present study, it was possible to visualize the germinal vesicle, the nucleolus, and the chromosome in the oocyte. After IVF, the nuclei, male and female pronuclei, polar bodies, nucleolus-associated chromatins, and chromosomes were successively observed in the zygote, the blastomere during cleavage, and the blastocyst until the hatching stage. The formation of male and female pronuclei, the process of their fusion, the appearance of the chromosome, and the commencement of the first cleavage of the zygote to form two blastomeres were all clearly demonstrated. The number of blastomeres was easily counted in the morula in spite of the compaction of the blastomeres. The present study indicates that the methods reported herein are useful for the determination of exact functional stages of the oocyte and the zygote during IVF and early embryo development, and thus contributes to the advancement of reproductive and developmental biomedical research.
在小鼠体外受精（IVF）及随后的早期胚胎发育过程中，对用光辉霉素或4',6-二脒基-2-苯基吲哚（DAPI）染色的卵母细胞和受精卵进行了荧光显微镜检查。光辉霉素和DAPI分别与DNA的G-C和A-T碱基对按化学计量结合。在本研究中，可以观察到卵母细胞中的生发泡、核仁及染色体。体外受精后，在受精卵、卵裂期的卵裂球及囊胚直至孵化阶段，依次观察到细胞核、雄原核和雌原核、极体、核仁相关染色质及染色体。雄原核和雌原核的形成、它们的融合过程、染色体的出现以及受精卵第一次卵裂形成两个卵裂球的起始过程均得到清晰展示。尽管卵裂球发生了致密化，但在桑葚胚中仍可轻松计数卵裂球的数量。本研究表明，本文报道的方法对于确定体外受精和早期胚胎发育过程中卵母细胞和受精卵的确切功能阶段很有用，从而有助于生殖和发育生物医学研究的进展。
