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首页> 外文期刊>Molecular reproduction and development >Suppression of connexin 43 and E-cadherin transcripts in in vitro derived bovine embryos following culture in vitro or in vivo in the homologous bovine oviduct.
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Suppression of connexin 43 and E-cadherin transcripts in in vitro derived bovine embryos following culture in vitro or in vivo in the homologous bovine oviduct.

机译：在同源牛输卵管中体外或体内培养后，在体外衍生的牛胚胎中抑制连接蛋白43和E-cadherin转录物。

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In this study, a combination of RNAi and endoscopic transfer to the oviduct of synchronized heifers has been used to investigate the effect of suppression of Cx43 and E-cadherin on the development, mRNA and protein expression of bovine blastocysts cultured in vitro or in vivo. In vitro matured and fertilized bovine zygotes were randomly assigned to one of four groups namely: Connexin43 dsRNA-injected (n = 790), E-cadherin dsRNA-injected (n = 775), water-injected (n = 774), and noninjected controls (n = 652). Following 2 days in vitro culture, 4- and 8-cell stage embryos from each treatment group were used for culture in vitro or in vivo. About half of the 4-8-cell stage embryos from each treatment group were transferred to the oviduct of synchronized heifers, while the remainder were further cultured in vitro. Embryos from in vivo culture were flushed from recipients on the fourth day post transfer (= Day 7 post insemination). Blastocyst stage embryos from both culture systems were used for mRNA and protein expression analysis. Irrespective of treatment or culture conditions, microinjection resulted in a decline in the proportion of embryos reaching the blastocyst stage. Significantly, lower blastocyst development was observed in E-cadherin and water-injected embryos following in vivo culture compared to the noninjected controls, while intermediate results were obtained following injection with Cx43 dsRNA. Both mRNA and protein products of the target genes were suppressed but the efficiency of suppression of the target genes varied depending on the initial level of transcript abundance, which is known to be greatly affected by the culture environment.
机译：在这项研究中，RNAi和内窥镜转移到同步母牛的输卵管中的组合已被用于研究抑制Cx43和E-钙粘蛋白对体外或体内培养的牛胚泡发育，mRNA和蛋白表达的影响。体外成熟和受精的牛受精卵随机分为四组之一：注射连接蛋白43 dsRNA（n = 790），注射E-钙粘蛋白dsRNA（n = 775），注水（n = 774）和不注射控制项（n = 652）。在体外培养2天后，将来自每个治疗组的4和8细胞期胚胎用于体外或体内培养。每个处理组的大约4-8个细胞阶段的胚胎转移到同步小母牛的输卵管中，其余的在体外进一步培养。移植后第四天（=授精后第7天）从受体冲洗掉体内培养物中的胚胎。来自两个培养系统的胚泡期胚胎用于mRNA和蛋白质表达分析。无论处理或培养条件如何，显微注射都会导致到达胚泡期的胚胎比例下降。显着地，与未注射的对照相比，在体内培养后在E-钙粘着蛋白和注射水的胚胎中观察到了较低的胚泡发育，而在注射Cx43 dsRNA之后获得了中间结果。靶基因的mRNA和蛋白质产物均被抑制，但是靶基因的抑制效率根据转录本丰度的初始水平而变化，已知转录本丰度最初受到培养环境的影响。

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