Fate of microinjected sperm components in the mouse oocyte and embryo
Fate of microinjected sperm components in the mouse oocyte and embryo
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DOI：
10.1017/s0967199400003889
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发表时间：
1997-11-01
期刊：
影响因子：
1.7
通讯作者：
Yanagimachi, R
中科院分区：
文献类型：
--
作者：
Cummins, JM;Wakayama, T;Yanagimachi, R
将用MitoTracker绿色FM荧光染料标记的完整小鼠精子或单独的小鼠精子尾部注射到小鼠卵母细胞中，并将细胞在体外培养长达5天。染色见中段线粒体、精子尾部粗纤维和精子穿孔器。完整的精子（或尾部注射分离的头部）诱导正常的胚胎发育。线粒体可以在胚胎中鉴定到4-细胞阶段，仍然与精子尾部相关。到了8细胞期，它们基本上消失了，只有少数胚胎（6/43）可以发现小块线粒体。轴丝元件可以确定卷曲在单个外部卵裂球迟至第5天囊胚。相比之下，线粒体以及尾部组件可以识别到5天后，精子尾部单独注射到未激活的卵母细胞，也在胚胎，逮捕前的8细胞阶段的发展。我们的结论是，标记的精子线粒体在正常分裂的胚胎中的消失不是由于荧光标记物的褪色或失活，而是一个事件，特别是与第二次细胞分裂周围的细胞周期活动。
Intact mouse sperm or mouse sperm tails alone, labelled with MitoTracker Green FM(R) fluorochrome, were injected into mouse oocytes and the cells cultured in vitro for up to 5 days. The dye stained midpiece mitochondria, the sperm tail coarse fibres and the sperm perforatorium. Intact sperm (or tails injected with separated heads) induced normal embryonic development. The mitochondria could be identified in embryos up to the 4-cell stage, remaining associated with the sperm tail. They largely disappeared by the 8-cell stage, when only a minority of embryos (6/43) could be found with small patches of mitochondria. Axonemal elements could be identified coiled up in single external blastomeres as late as day 5 blastocysts. By contrast, mitochondria as well as tail components could be identified up to 5 days after injection of sperm tails alone into non-activated oocytes and also in embryos that arrested development before the 8-cell stage. We conclude that disappearance of the labelled sperm mitochondria in normally cleaving embryos is not due to fading or inactivation of the fluorochrome marker, but is rather an event specifically tied to cell cycle activities around the second cell division.