Polarization of blastomeres in the cleaving rabbit embryo.
Polarization of blastomeres in the cleaving rabbit embryo.
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DOI：
10.1002/jez.1402560111
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发表时间：
1990-10
期刊：
影响因子：
--
通讯作者：
C. Ziomek;C. Chatot;C. Manes
中科院分区：
文献类型：
--
作者：
C. Ziomek;C. Chatot;C. Manes
细胞极化被认为是啮齿动物胚胎中两种细胞系分化分化形成囊胚的关键事件。本研究旨在确定兔胚胎在囊胚形成之前是否表现出细胞极化，并确定极化首次出现的胚胎阶段。通过观察FITC-Con A在兔胚胎三个阶段离体卵裂球上的结合模式来测定其极性。对游离细胞的扫描电镜观察证实了荧光结果。38- 66细胞兔胚胎中，51%的卵裂球具有强烈的FITC-Con - A结合极和单极微绒毛。在17- 34细胞阶段，只有2%的卵裂球出现类似的极化，而在8- 16细胞阶段则没有出现类似的极化。此外，在兔胚胎分离前试图去除粘蛋白外壳和透明带时，发现这些外壳的蛋白酶敏感性在38- 66细胞阶段也发生了变化。在此之前，虽然在0.5% pronase中30分钟后粘蛋白包被消失，但即使部分去除，也需要大约1.5-2.5小时的pronase。在38- 66细胞阶段，蛋白酶在10分钟内溶解粘蛋白外壳，在20分钟内溶解透明带。在所有检测阶段，该带对0.1%蛋白酶K均具有抗性。
Cellular polarization is believed to be a crucial event in the differentiative divergence of the two cell lineages leading to the blastocyst in rodent embryos. This study was undertaken to determine if rabbit embryos exhibited cellular polarization prior to blastocyst formation and to define the embryonic stage at which polarization was first apparent. Polarity was assayed by observation of the pattern of binding of FITC-Con A to dissociated blastomeres from three stages of rabbit embryos. Scanning electron microscopy on the dissociated cells confirmed the fluorescence results. Fifty-one percent of blastomeres in 38- to 66-cell rabbit embryos exhibited an intense pole of FITC-Con A binding and a single pole of microvilli. Only 2% of blastomeres at the 17- to 34-cell stage were similarly polarized and none were polarized at the 8- to 16-cell stage. In addition, during attempts to remove the mucin coat and zona pellucida from the rabbit embryos prior to their dissociation, it was found that the protease sensitivity of these coats also changed at the 38- to 66-cell stage. Prior to this time, although the mucin coat disappeared after 30 min in 0.5% pronase, the zona required approximately 1.5-2.5 hr in pronase for even partial removal. At the 38- to 66-cell stage, pronase dissolved the mucin coat within 10 min and the zona pellucida within 20 min. The zona was resistant to 0.1% proteinase K at all stages examined.