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小鼠胚胎细胞核移植到去核卵母细胞后核重塑与后续发育之间的关系。

Relationship between nuclear remodeling and subsequent development of mouse embryonic nuclei transferred to enucleated oocytes.

作者信息

Cheong H T, Takahashi Y, Kanagawa H

机构信息

Department of Theriogenology, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

出版信息

Mol Reprod Dev. 1994 Feb;37(2):138-45. doi: 10.1002/mrd.1080370204.

Abstract

The present study was conducted to examine the relationship between nuclear remodeling and subsequent embryonic development in nuclear transplant mouse embryos. Metaphase II oocytes were enucleated without staining and fused with transferred donor nuclei from two-, four-, or eight-cell embryos. Fusion and oocyte activation were performed by means of electric fields. High rates of enucleation (89.1%), fusion (88.0-91.6%), and activation (95.2-96.9%) were obtained using this system. Nuclear remodeling was characterized by premature chromosome condensation (PCC), followed by various pronuclear-like formations upon oocyte activation. Development to blastocysts was obtained from both PCC (17.9%) and non-PCC (NPCC; 52.9%) embryos fused with the two-cell nuclei. However, development to term was obtained only in PCC embryos with a single pronucleus-like structure and a polar body (12.5%). In vitro development of nuclear transplant embryos with four- and eight-cell nuclei was limited. All the NPCC embryos examined had tetraploid chromosome constitutions, but chromosome constitutions of PCC embryos varied. Only 37.5% of the PCC embryos had diploid chromosome constitutions. The results indicated that the development of nuclear transplant embryos is affected by the types of nuclear remodeling and that oocyte activation in relation to their chromosome constitutions. The results also indicated that the PCC of the donor nucleus in nonactivated cytoplasm is important for the development of the nuclear transplant embryos.

摘要

本研究旨在探讨核移植小鼠胚胎中核重塑与后续胚胎发育之间的关系。对处于减数分裂中期II的卵母细胞进行去核操作,不进行染色,并与来自二细胞、四细胞或八细胞胚胎的移植供体细胞核融合。通过电场进行融合和卵母细胞激活。使用该系统获得了较高的去核率(89.1%)、融合率(88.0 - 91.6%)和激活率(95.2 - 96.9%)。核重塑的特征是过早染色体凝聚(PCC),随后在卵母细胞激活时出现各种类原核形成。与二细胞核融合的PCC胚胎(17.9%)和非PCC(NPCC;52.9%)胚胎均发育至囊胚。然而,只有具有单个类原核结构和一个极体的PCC胚胎发育至足月(12.5%)。具有四细胞和八细胞核的核移植胚胎的体外发育受到限制。所有检测的NPCC胚胎都具有四倍体染色体组成,但PCC胚胎的染色体组成各不相同。只有37.5%的PCC胚胎具有二倍体染色体组成。结果表明,核移植胚胎的发育受核重塑类型的影响,并且与它们的染色体组成相关的卵母细胞激活也有影响。结果还表明,未激活细胞质中供体细胞核的PCC对核移植胚胎的发育很重要。

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