A simplified one-step nuclear transfer procedure alters the gene expression patterns and developmental potential of cloned porcine embryos.
A simplified one-step nuclear transfer procedure alters the gene expression patterns and developmental potential of cloned porcine embryos.
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DOI：
10.4142/jvs.2014.15.1.73
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发表时间：
2014
影响因子：
1.8
通讯作者：
Park JI
中科院分区：
文献类型：
--
作者：
Park SK;Roh S;Park JI
各种用于哺乳动物物种的体细胞核移植（SCNT）技术已经开发出来，以调整物种特异性程序以适应物种之间卵母细胞相关的差异。物种特异性SCNT方案可能导致影响胚胎发育和妊娠的重要基因的不同表达水平。在本研究中，猪卵母细胞用去焦碱处理，使其具有突出的遗传物质，促进去核。去核和供体细胞注射用单个移液管（简化一步SCNT； SONT）同时进行，或分别用不同的移液管（常规两步SCNT； CTNT）作为对照程序。两组囊胚体外培养后，采用实时定量聚合酶链反应分析OCT4、NANOG、EOMES、CDX2、GLUT-1、PolyA、HSP70等发育重要基因的表达水平。与CTNT相比，SONT提高了囊胚分期的发育率以及CDX2、EOMES和HSP70的表达水平。已知表达升高的基因影响滋养外胚层的形成和热应激诱导的阻滞。这些结果表明，我们的SONT技术改善了SCNT猪胚胎的发育，并增加了胎盘形成和应激诱导停止的重要基因的表达。
Various somatic cell nuclear transfer (SCNT) techniques for mammalian species have been developed to adjust species-specific procedures to oocyte-associated differences among species. Species-specific SCNT protocols may result in different expression levels of developmentally important genes that may affect embryonic development and pregnancy. In the present study, porcine oocytes were treated with demecolcine that facilitated enucleation with protruding genetic material. Enucleation and donor cell injection were performed either simultaneously with a single pipette (simplified one-step SCNT; SONT) or separately with different pipettes (conventional two-step SCNT; CTNT) as the control procedure. After blastocysts from both groups were cultured in vitro, the expression levels of developmentally important genes (OCT4, NANOG, EOMES, CDX2, GLUT-1, PolyA, and HSP70) were analyzed by real-time quantitative polymerase chain reaction. Both the developmental rate according to blastocyst stage as well as the expression levels CDX2, EOMES, and HSP70 were elevated with SONT compared to CTNT. The genes with elevated expression are known to influence trophectoderm formation and heat stress-induced arrest. These results showed that our SONT technique improved the development of SCNT porcine embryos, and increased the expression of genes that are important for placental formation and stress-induced arrest.
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影响因子：
4.6
作者：
Strumpf, D;Mao, CA;Rossant, J
通讯作者：
Rossant, J
影响因子：
6.1
作者：
Mitalipov, S. M.;Zhou, Q.;Wolf, D. P.
通讯作者：
Wolf, D. P.
影响因子：
2.8
作者：
Costa-Borges, Nuno;Teresa Paramio, Maria;Ibanez, Elena
通讯作者：
Ibanez, Elena
DOI：
10.1089/clo.2004.6.294
发表时间：
2004-09-01
期刊：
CLONING AND STEM CELLS
影响因子：
--
作者：
Fujimura, T;Kurome, M;Nagashima, H
通讯作者：
Nagashima, H
影响因子：
2.5
作者：
Isom, S. Clay;Lai, Liangxue;Rucker, Edmund B., III
通讯作者：
Rucker, Edmund B., III