Somatic and embryonic cell nucleus transfer into intact and enucleated immature mouse oocytes
Somatic and embryonic cell nucleus transfer into intact and enucleated immature mouse oocytes
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DOI：
10.1093/humrep/17.8.2160
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发表时间：
2002-08-01
影响因子：
6.1
通讯作者：
Tesarik, J
中科院分区：
文献类型：
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作者：
Fulka, J;Martinez, F;Tesarik, J
背景：我们研究的目的是评估胚胎或体细胞单倍体化与完整或去核的未成熟卵母细胞融合后体外培养的可能性。当胚胎或体细胞细胞核融合到完整或去核的未成熟卵母细胞时，不会发生过早的染色体凝聚，而二丁基环AMP （dbcAMP）会阻止其成熟。然而，dbcAMP的存在允许在体细胞细胞核中完成DNA复制。方法与结果：重组细胞从dbcAMP阻滞中释放后，染色体凝聚。当体细胞或胚胎核被引入完整的未成熟的有减数分裂能力的卵母细胞并随后培养时，它们的染色体与减数分裂染色体聚集在一个共同的纺锤上，并进行减数分裂样分裂，根据挤出的第一极体的存在来判断。将胚胎细胞核导入未成熟的有减数分裂能力的卵母细胞的细胞质中，约75%的重建细胞中有极体挤出，但几乎所有细胞的中期板都异常。当体细胞细胞核插入上述细胞质时，极体只在非常特殊的情况下被挤出，在这些细胞中染色体排列在流产中期板上。结论：我们的研究结果表明，将体细胞细胞核引入未成熟卵母细胞减数分裂细胞质时，细胞核无法进行减数分裂（单倍体化）。
BACKGROUND: The aim of our study was to evaluate the possibility of embryonic or somatic cell haploidization after fusion with intact or enucleated immature oocytes which were subsequently cultured in vitro. Embryonic or somatic cell nuclei do not undergo premature chromosome condensation when fused to intact or enucleated immature oocytes whose maturation is prevented by dibutyryl cyclic AMP (dbcAMP). The presence of dbcAMP permits, however, the completion of DNA replication in somatic cell nuclei. METHODS AND RESULTS: The chromosomes condensed when the reconstructed cells were released from the dbcAMP block. When somatic or embryonic nuclei were introduced into intact immature meiotically competent oocytes and subsequently cultured their chromosomes assembled on a common spindle with meiotic chromosomes and proceeded through the meiotic-like division, judged according to the presence of the first polar body extruded. When embryonic cell nuclei were introduced into cytoplasts obtained from immature meiotically competent oocytes, polar bodies were extruded in about 75% of reconstructed cells but the metaphase plates were abnormal in almost all cases. When somatic cell nuclei were inserted into the above cytoplasts, polar bodies were extruded only very exceptionally and in these cells chromosomes were arranged in abortive metaphase plates. CONCLUSIONS: Our results suggest that somatic cell nuclei are unable to proceed through the reduction division (haploidization) when introduced into an immature oocyte meiotic cytoplasm.