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牛囊胚的全球基因表达概要:环境条件的反映者和发育能力的预测者。

The global gene expression outline of the bovine blastocyst: reflector of environmental conditions and predictor of developmental capacity.

机构信息

Institute of Animal Sciences, Animal Breeding, University of Bonn, Endenicher Allee 15, 53115, Bonn, Germany.

Animal Reproduction and Biotechnology Laboratory, Department of Biomedical Sciences, Colorado State University, 3105 Rampart Rd, CO, 80521, Fort Collins, USA.

出版信息

BMC Genomics. 2021 Jun 3;22(1):408. doi: 10.1186/s12864-021-07693-0.

Abstract

BACKGROUND

Morphological evaluation of embryos has been used to screen embryos for transfer. However, the repeatability and accuracy of this method remains low. Thus, evaluation of an embryo's gene expression signature with respect to its developmental capacity could provide new opportunities for embryo selection. Since the gene expression outline of an embryo is considered as an aggregate of its intrinsic characteristics and culture conditions, we have compared transcriptome profiles of in vivo and in vitro derived blastocysts in relation to pregnancy outcome to unravel the discrete effects of developmental competence and environmental conditions on bovine embryo gene expression outlines. To understand whether the gene expression patterns could be associated with blastocyst developmental competency, the global transcriptome profile of in vivo (CVO) and in vitro (CVT) derived competent blastocysts that resulted in pregnancy was investigated relative to that of in vivo (NVO) and in vitro (NVT) derived blastocysts which did not establish initial pregnancy, respectively while to unravel the effects of culture condition on the transcriptome profile of embryos, the transcriptional activity of the CVO group was compared to the CVT group and the NVO group was compared to the NVT ones.

RESULTS

A total of 700 differentially expressed genes (DEGs) were identified between CVO and NVO blastocysts. These gene transcripts represent constitutive regions, indel variants, 3'-UTR sequence variants and novel transcript regions. The majority (82%) of these DEGs, including gene clusters like ATP synthases, eukaryotic translation initiation factors, ribosomal proteins, mitochondrial ribosomal proteins, NADH dehydrogenase and cytochrome c oxidase subunits were enriched in the CVO group. These DEGs were involved in pathways associated with glycolysis/glycogenesis, citrate acid cycle, pyruvate metabolism and oxidative phosphorylation. Similarly, a total of 218 genes were differentially expressed between CVT and NVT groups. Of these, 89%, including TPT1, PDIA6, HSP90AA1 and CALM, were downregulated in the CVT group and those DEGs were overrepresented in pathways related to protein processing, endoplasmic reticulum, spliceasome, ubiquitone mediated proteolysis and steroid biosynthesis. On the other hand, although both the CVT and CVO blastocyst groups resulted in pregnancy, a total of 937 genes were differential expressed between the two groups. Compared to CVO embryos, the CVT ones exhibited downregulation of gene clusters including ribosomal proteins, mitochondrial ribosomal protein, eukaryotic translation initiation factors, ATP synthases, NADH dehydrogenase and cytochrome c oxidases. Nonetheless, downregulation of these genes could be associated with pre and postnatal abnormalities observed after transfer of in vitro embryos.

CONCLUSION

The present study provides a detailed inventory of differentially expressed gene signatures and pathways specifically reflective of the developmental environment and future developmental capacities of bovine embryos suggesting that transcriptome activity observed in blastocysts could be indicative of further pregnancy success but also adaptation to culture environment.

摘要

背景

胚胎形态学评估已被用于筛选可移植胚胎。然而,这种方法的重复性和准确性仍然较低。因此,评估胚胎的基因表达特征及其发育能力可能为胚胎选择提供新的机会。由于胚胎的基因表达轮廓被认为是其内在特征和培养条件的总和,我们比较了体内和体外来源的囊胚的转录组谱与妊娠结局,以揭示发育能力和环境条件对牛胚胎基因表达轮廓的离散影响。为了了解基因表达模式是否与囊胚发育能力相关,我们研究了体内(CVO)和体外(CVT)来源的有能力的囊胚的整体转录组谱,这些囊胚导致妊娠,而体内(NVO)和体外(NVT)来源的囊胚则未建立初始妊娠。此外,为了揭示培养条件对胚胎转录组谱的影响,我们比较了 CVO 组与 CVT 组,NVO 组与 NVT 组的转录活性。

结果

CVO 和 NVO 囊胚之间共鉴定出 700 个差异表达基因(DEGs)。这些基因转录本代表组成型区域、插入缺失变体、3'-UTR 序列变体和新的转录本区域。这些 DEGs 中的大多数(82%),包括 ATP 合酶、真核翻译起始因子、核糖体蛋白、线粒体核糖体蛋白、NADH 脱氢酶和细胞色素 c 氧化酶亚基等基因簇,在 CVO 组中富集。这些 DEGs 参与与糖酵解/糖异生、柠檬酸循环、丙酮酸代谢和氧化磷酸化相关的途径。同样,CVT 和 NVT 组之间共鉴定出 218 个差异表达基因。其中,89%,包括 TPT1、PDIA6、HSP90AA1 和 CALM,在 CVT 组中下调,这些 DEGs 在与蛋白质加工、内质网、剪接体、泛素介导的蛋白水解和类固醇生物合成相关的途径中过表达。另一方面,尽管 CVT 和 CVO 囊胚组均导致妊娠,但两组之间共鉴定出 937 个差异表达基因。与 CVO 胚胎相比,CVT 胚胎中包括核糖体蛋白、线粒体核糖体蛋白、真核翻译起始因子、ATP 合酶、NADH 脱氢酶和细胞色素 c 氧化酶等基因簇下调。然而,这些基因的下调可能与体外胚胎移植后观察到的产前和产后异常有关。

结论

本研究提供了一个详细的差异表达基因特征和途径清单,这些特征和途径专门反映了牛胚胎的发育环境和未来的发育能力,表明囊胚中观察到的转录组活性可能预示着进一步的妊娠成功,但也可能预示着对培养环境的适应。

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