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卵母细胞细胞质体积对孤雌激活、胞浆内单精子注射或体细胞核移植后胚胎发育的影响。

Effect of volume of oocyte cytoplasm on embryo development after parthenogenetic activation, intracytoplasmic sperm injection, or somatic cell nuclear transfer.

作者信息

Sayaka Wakayama, Satoshi Kishigami, Van Thuan Nguyen, Hiroshi Ohta, Takafusa Hikichi, Eiji Mizutani, Thuy Bui Hong, Masashi Miyake, Teruhiko Wakayama

机构信息

Laboratory for Genomic Reprogramming, Center for Developmental Biology, RIKEN, Minatojima-minamimachi Chuo-ku, Kobe, Japan.

出版信息

Zygote. 2008 Aug;16(3):211-22. doi: 10.1017/S0967199408004620. Epub 2008 Jun 26.

Abstract

Animal cloning methods are now well described and are becoming routine. Yet, the frequency at which live cloned offspring are produced remains below 5%, irrespective of the nuclear donor species or cell type. One possible explanation is that the reprogramming factor(s) of each oocyte is insufficient or not properly adapted for the receipt of a somatic cell nucleus, because it is naturally prepared only for the receipt of a gamete. Here, we have increased the oocyte volume by oocyte fusion and examined its subsequent development. We constructed oocytes with volumes two to nine times greater than the normal volume by the electrofusion or mechanical fusion of intact and enucleated oocytes. We examined their in vitro and in vivo developmental potential after parthenogenetic activation, intracytoplasmic sperm injection (ICSI) and somatic cell nuclear transfer (SCNT). When the fused oocytes were activated parthenogenetically, most developed to morulae or blastocysts, regardless of their original size. Diploid fused oocytes were fertilized by ICSI and developed normally and after embryo transfer, we obtained 12 (4-15%) healthy and fertile offspring. However, enucleated fused oocytes could not support the development of mice cloned by SCNT. These results suggest that double fused oocytes have normal potential for development after fertilization, but oocytes with extra cytoplasm do not have enhanced reprogramming potential.

摘要

动物克隆方法现已得到充分描述并日趋常规化。然而,无论核供体物种或细胞类型如何,克隆后代存活出生的频率仍低于5%。一种可能的解释是,每个卵母细胞的重编程因子不足或未适当适配于接受体细胞核,因为它天然仅准备好接受配子。在此,我们通过卵母细胞融合增加了卵母细胞体积,并研究了其随后的发育情况。我们通过完整和去核卵母细胞的电融合或机械融合构建了体积比正常体积大两到九倍的卵母细胞。我们研究了孤雌激活、胞浆内单精子注射(ICSI)和体细胞核移植(SCNT)后它们的体外和体内发育潜能。当融合卵母细胞被孤雌激活时,大多数发育至桑葚胚或囊胚,无论其原始大小如何。二倍体融合卵母细胞经ICSI受精并正常发育,胚胎移植后,我们获得了12只(4 - 15%)健康且可育的后代。然而,去核融合卵母细胞无法支持通过SCNT克隆的小鼠的发育。这些结果表明,双融合卵母细胞在受精后具有正常的发育潜能,但具有额外细胞质的卵母细胞没有增强的重编程潜能。

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