Differential effect of recipient cytoplasm for microtubule organization and preimplantation development in rat reconstituted embryos with two‐cell embryonic cell nuclear transfer
Differential effect of recipient cytoplasm for microtubule organization and preimplantation development in rat reconstituted embryos with two‐cell embryonic cell nuclear transfer
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受体细胞质对双细胞胚胎细胞核移植大鼠重建胚胎微管组织和植入前发育的差异作用
DOI：
10.1002/mrd.20083
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发表时间：
2004
影响因子：
2.5
通讯作者：
E. Sato
中科院分区：
文献类型：
--
作者：
T. Shinozawa;E. Mizutani;I. Tomioka;M. Kawahara;H. Sasada;H. Matsumoto;E. Sato
在本研究中，我们研究了去核受精卵，MII卵母细胞，孤雌激活的卵母细胞在原核阶段（孤雌生殖PNs）作为受体细胞质的大鼠胚胎细胞核移植的发育能力。去核受精卵作为接受双细胞核的受体细胞质允许发育成囊胚，而用MII卵母细胞和孤雌生殖PN重建的胚胎的发育在双细胞阶段被阻止。先前在大鼠两细胞胚胎中的观察表明，微管的分布与两细胞停滞有关。因此，我们还使用免疫荧光检查了微管的分布。在核移植到去核受精卵后的两细胞阶段，微管在间期均匀分布在细胞质中，并且在两细胞至四细胞阶段的分裂胚胎中观察到正常的有丝分裂纺锤体。相反，MII卵母细胞和孤雌生殖PN重建的胚胎显示异常的微管组织。在去核受精卵中，纤维状微管均匀分布在细胞质中。与此相反，密集的微管定位在皮质下区域的细胞质和强烈的免疫荧光强度观察质膜，而非常弱的强度检测到去核MII卵母细胞的中央部分。在去核孤雌生殖PN中，高密度和纤维微管分别分布在皮质下和中央区域。去核前的孤雌生殖PN在中央细胞质中的微管免疫荧光强度也低于合子。总之，本研究的结果表明，对于大鼠二细胞胚胎细胞核移植，受精卵细胞质比MII卵母细胞和孤雌生殖PN更适合作为受体，以发育超过四细胞阶段。此外，微管组织参与重建胚胎的发育，以克服两细胞停滞。摩尔Reprod. Dev. 68：313-318，2004.© 2004 Wiley利斯公司
In the present study, we examined the developmental ability of enucleated zygotes, MII oocytes, and parthenogenetically activated oocytes at pronuclear stages (parthenogenetic PNs) as recipient cytoplasm for rat embryonic cell nuclear transfer. Enucleated zygotes as recipient cytoplasm receiving two‐cell nuclei allowed development to blastocysts, whereas the development of embryos reconstituted with MII oocytes and parthenogenetic PNs was arrested at the two‐cell stage. Previous observations in rat two‐cell embryos suggested that the distribution of microtubules is involved in two‐cell arrest. Therefore, we also examined the distribution of microtubules using immunofluorescence. At the two‐cell stage after nuclear transfer into enucleated zygotes, microtubules were distributed homogeneously in the cytoplasm during interphase, and normal mitotic spindles were observed in cleaving embryos from the two‐ to four‐cell stage. In contrast, embryos reconstituted with MII oocytes and parthenogenetic PNs showed aberrant microtubule organization. In enucleated zygotes, fibrous microtubules were distributed homogeneously in the cytoplasm. In contrast, dense microtubules were localized at the subcortical area in the cytoplasm and strong immunofluorescence intensity was observed at the plasma membrane, while very weak intensity was detected in the central part of enucleated MII oocytes. In enucleated parthenogenetic PNs, high‐density and fibrous microtubules were distributed in the subcortical and central areas, respectively. Pre‐enucleated parthenogenetic PNs also showed lower intensity of microtubule immunofluorescence in the central cytoplasm than zygotes. In conclusion, the results of the present study showed that zygote cytoplasm is better as recipient than MII oocyte and parthenogenetic PNs for rat two‐cell embryonic cell nuclear transfer to develop beyond four‐cell stage. Furthermore, microtubule organization is involved in the development of reconstituted embryos to overcome the two‐cell arrest. Mol. Reprod. Dev. 68: 313–318, 2004. © 2004 Wiley‐Liss, Inc.
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影响因子：
4.6
作者：
D. Worrad;Prahlad T. Ram;R. Schultz
通讯作者：
D. Worrad;Prahlad T. Ram;R. Schultz
影响因子：
13.3
作者：
Bavister, BD
通讯作者：
Bavister, BD
影响因子：
3.6
作者：
Meng, L;Ely, JJ;Wolf, DP
通讯作者：
Wolf, DP
影响因子：
2.7
作者：
RAM, PT;SCHULTZ, RM
通讯作者：
SCHULTZ, RM