Development of embryos from in vitro ovulated and fertilized oocytes of the quail (Coturnix coturnix japonica)
Development of embryos from in vitro ovulated and fertilized oocytes of the quail (Coturnix coturnix japonica)
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DOI：
10.1002/jez.10096
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发表时间：
2002-05-01
影响因子：
--
通讯作者：
Perry, MM
中科院分区：
文献类型：
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作者：
Olszanska, B;Stepinska, U;Perry, MM
研究了鹌鹑卵母细胞体外受精后胚胎的发育情况。在立体显微镜下观察，约40%的标本经过18-20小时的孵育，已发生裂解，达到IV-VI期。然而，在荧光显微镜下观察，这些胚胎中只有36%含有正常的dapi染色的细胞核；其余64%显示形态正常的卵裂模式，不含细胞核。对照未受精卵母细胞，在体外排卵和培养的同时，有时也达到形态正确的IV-VI期，但它们的“卵裂球”总是无核的。因此，在评估培养发育时，监测早期禽类胚胎是否存在细胞核是可取的。这一结果首次证明了鹌鹑生发盘在没有核分裂的情况下也可以发生细胞质分裂，表明了鸟类卵浆母性信息的存在及其意义。这种现象在海胆和青蛙身上也很常见。这表明母体信息在早期发育中的作用。本文提出的体外方法在体外条件下将排卵和受精的步骤与早期卵裂阶段联系起来，可用于研究鸟类的受精和分化机制，以及通过DNA注射或应用外源携带DNA的精子获得转基因鸟类。[j] .浙江大学学报（自然科学版）。580 - 586年,2002年。(C) 2002 Wiley-Liss, Inc。
The development of quail embryos obtained after in vitro fertilization of oocytes ovulated in vitro was investigated. About 40% of the specimens,, after 18-20 hr of incubation, had undergone cleavage to reach stages IV-VI when viewed under a stereo microscope. However, only 36% of these embryos contained normal, DAPI-stained nuclei when observed under a fluorescent microscope; the other 64% showing a morphologically normal cleavage pattern did not contain nuclei. Control unfertilized oocytes, ovulated in vitro and cultured for the same time, also sometimes attained the morphologically correct stages IV-VI but their "blastomeres" were always devoid of nuclei. Therefore, it is advisable to monitor early avian embryos for the presence of nuclei when assessing development in culture.The results demonstrate, for the first time, that cytoplasmic segmentation can occur in the absence of nuclear divisions in the germinal disc of the quail and show the existence and significance of ooplasmic maternal information in birds. This phenomenon is also known for sea urchin and frogs. It is indicative of the role of maternal information in early development. The in vitro method presented here links the steps of ovulation and fertilization with the early cleavage stages under in vitro conditions and may be useful in studying mechanisms of fertilization and differentiation in birds as well as in obtaining transgenic birds by DNA injection or application of foreign, DNA-carrying sperm. J. Exp. Zool. 292.-580-586, 2002. (C) 2002 Wiley-Liss, Inc.