Temporal expression of transcripts related to embryo quality in bovine embryos cultured from the two-cell to blastocyst stage in vitro or in vivo
Temporal expression of transcripts related to embryo quality in bovine embryos cultured from the two-cell to blastocyst stage in vitro or in vivo
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DOI：
10.1002/mrd.20677
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发表时间：
2007-08-01
影响因子：
2.5
通讯作者：
Lonergan, Patrick
中科院分区：
文献类型：
--
作者：
Corcoran, Deirdre;Rizos, Dimitrios;Lonergan, Patrick
受精后胚胎培养环境对胚胎中的基因表达模式具有显著影响，并且人们普遍认为，体外培养的牛胚胎的质量低于体内培养的牛胚胎。本研究的目的是研究已知的几种转录和翻译因子的mRNA丰度的时间变化，这些转录和翻译因子在体外和体内培养后产生的囊胚之间存在差异。从两个体外培养系统SOF 1或SOF 2中回收了5个发育阶段的胚胎：2- 4-细胞、8-细胞、16-细胞、桑椹胚和囊胚。从超数排卵和人工授精的小母牛产生体内胚胎，并在授精后约40小时或3、4、5和7天回收。体外成熟、体外受精和在母羊输卵管中培养也产生囊胚。使用定量实时PCR对FOX 03 A、EEF 1G、HMG 2和REA的相对转录丰度进行分析。无论培养环境如何，每个转录物遵循大致相同的一般表达模式，其中相对丰度从2-至4-细胞期到8-细胞期急剧降低，并且从桑椹胚到囊胚期增加（P < 0.05）。在发育的2-和16-细胞阶段之间未观察到GNBL 2的转录物。在2- 4-细胞中相对高的表达表明这些转录物最有可能是在卵母细胞生长和最终成熟过程中产生的母体来源。一个文化诱导的转录和翻译因子的mRNA丰度的变化是显而易见的，不仅在体内和体外培养环境之间，但也在不同的体外培养系统之间产生的胚胎。
The post-fertilization embryo culture environment can have a dramatic effect on the pattern of gene expression in the embryo and it is widely acknowledged that bovine embryos derived from in vitro culture are of inferior quality to those derived in vivo. The objective of this study was to examine temporal variation in the mRNA abundance of several transcription and translation factors known to differ between blastocysts produced following culture in vitro and in vivo. Embryos were recovered from two in vitro culture systems SOF1 or SOF2 at five developmental stages: 2- to 4-cell, 8-cell, 16-cell, morula, and blastocyst. In vivo embryos were produced from superovulated and artificially inseminated heifers and recovered at approximately 40 hr or 3, 4, 5, and 7 days postinsemination. Blastocysts were also produced following in vitro maturation, in vitro fertilization and culture in the ewe oviduct. Analysis of relative transcript abundance for FOX03A, EEF1G, HMG2, and REA was performed using quantitative real-time PCR. Irrespective of culture environment each transcript followed, approximately the same general pattern of expression where relative abundance decreased dramatically from the 2- to 4-cell stage to 8-cell stage and increased from the morula to blastocyst stage (P < 0.05). Transcripts for GNBL2 were not observed between the 2- and 16-cell stage of development. Relatively high expression at the 2- to 4-cell indicated that these transcripts are most likely of maternal origin produced in the oocyte during growth and final maturation. A culture-induced change in mRNA abundance of transcription and translation factors was evident in embryos that were produced not only between in vivo and in vitro culture environments but also between different in vitro culture systems.