Nucleoli From Growing Oocytes Support the Development of Enucleolated Full-Grown Oocytes in the Pig
Nucleoli From Growing Oocytes Support the Development of Enucleolated Full-Grown Oocytes in the Pig
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DOI：
10.1002/mrd.21126
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发表时间：
2010-02-01
影响因子：
2.5
通讯作者：
Miyano, Takashi
中科院分区：
文献类型：
--
作者：
Kyogoku, Hirohisa;Ogushi, Sugako;Miyano, Takashi
最近的研究表明，母体核仁对胚胎发育至关重要。生长中的卵母细胞的核仁形态与完全生长的卵母细胞的核仁形态不同。本研究旨在探讨生长期卵母细胞的核仁替代成熟期卵母细胞的核仁对胚胎发育的支持作用。分别从小腔卵泡（0.6-1.0 mm）和大腔卵泡（4-5 mm）中收集生长（直径约100 μ m）和完全生长的猪卵母细胞（120 μ m）。通过显微操作从完全生长的卵母细胞中吸出核仁，并将所得的去核卵母细胞成熟至中期11;然后将来自完全生长和生长的卵母细胞的核仁注射到卵母细胞中。在去核过程中，生长期卵母细胞的染色质随核仁一起吸出。因此，用放线菌素D处理生长中的卵母细胞以从其核仁释放染色质，收集核仁并转移到去核和成熟的完全生长的卵母细胞中。假手术卵母细胞经电刺激激活后，原核内有核仁形成。去核的卵母细胞，已注射从无论是完全成长或成长，但是，没有形成任何核仁的原核。没有去核卵母细胞发育成囊胚，而去核卵母细胞注射来自完全生长的卵母细胞（15%）或生长的卵母细胞（18%）发育成囊胚。这些结果表明，在早期胚胎发育过程中，生长期卵母细胞的核仁可以替代成熟期卵母细胞的核仁。
Recent research has shown that the maternal nucleolus is essential for embryonic development. The morphology of the nucleolus in growing oocytes differs from that in full-grown oocytes. We determined the ability of nucleoli from growing oocytes to substitute for nucleoli of full-grown oocytes in terms of supporting embryonic development in this study. Growing (around 100 mu m in diameter) and full-grown porcine oocytes (120 mu m) were collected from small (0.6-1.0 mm) and large antral follicles (4-5 mm), respectively. The nucleolus was aspirated from full-grown oocytes by micromanipulation, and the resulting enucleolated oocytes were matured to metaphase 11; the nucleoli originating from full-grown and growing oocytes were then injected into the oocytes. The Chromatin of growing oocytes was aspirated with the nucleolus during the enucleolation process. Growing oocytes were thus treated with actinomycin D to release the chromatin from their nucleoli, and the nucleoli were collected and transferred to the enucleolated and matured full-grown oocytes. After activation by electro-stimulation, nucleoli were formed in pronuclei of sham-operated oocytes. Enucleolated oocytes that had been injected with nucleoli from either full-grown or growing, however, did not form any nucleoli in the pronuclei. No enucleolated oocytes developed to blastocysts, whereas enucleolated oocytes injected with nucleoli from full-grown oocytes (15%) or growing oocytes (18%) developed to blastocysts. These results indicate that the nucleoli from growing oocytes can substitute for nucleoli from full-grown oocytes during early embryonic development.