Influence of Sex on Basal and Dickkopf-1 Regulated Gene Expression in the Bovine Morula.
Influence of Sex on Basal and Dickkopf-1 Regulated Gene Expression in the Bovine Morula.
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DOI：
10.1371/journal.pone.0133587
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Hansen PJ
中科院分区：
文献类型：
--
作者：
Denicol AC;Leão BC;Dobbs KB;Mingoti GZ;Hansen PJ
早在胚胎着床前，性别就会影响哺乳动物胚胎发育的功能。当前的目标有两个目标。首先是确定奶牛发育阶段雌性和雄性胚胎基因表达差异的程度和性质。第二个目标是确定DKK1（一种已知能改变囊胚分化的分子）是否会对雌性和雄性桑葚胚的基因表达产生不同的影响。在实验1中，在受精后第5天用DKK1处理雌性和雄性胚胎。处理24 h后采集桑葚，每20个为一组进行微阵列分析，RNA进行基因表达分析。男女差异表达基因共有662个，其中128个基因在两性间存在≥1.5倍的倍变。在女性中上调的基因中，49.5%位于X染色体。功能分析预测，雌性胚胎的细胞存活率更高。实验2涉及类似的设计，除了先前报道的受性别，DKK1或相互作用影响的12个基因的转录本通过定量聚合酶链反应进行定量。实验1中受性别影响的所有基因的表达在实验2中也以类似的方式受到影响。相比之下，DKK1对基因表达的影响在实验2中基本上不可重复。Hippo信号基因AMOT是例外，它被DKK1抑制。在实验3中，未分选精子受精产生的胚胎在第5天用DKK1处理，并在授精后第5、6和7天检测CDX2、GATA6和NANOG转录本的丰度。DKK1对这三个基因的表达均无影响。综上所述，早在桑葚胚期，雌性和雄性牛胚胎的基因表达模式就有所不同，这在很大程度上是由于雌性X染色体上的基因表达所致。雌性和雄性胚胎之间的差异基因表达可能是雌性胚胎对细胞死亡信号的抵抗力增强以及雌性和雄性胚胎对母体环境变化的反应差异的基础。
Sex affects function of the developing mammalian embryo as early as the preimplantation period. There were two goals of the current objective. The first was to determine the degree and nature of differences in gene expression between female and male embryos in the cow at the morula stage of development. The second objective was to determine whether DKK1, a molecule known to alter differentiation of the blastocyst, would affect gene expression differently for female and male morulae. In Experiment 1, female and male embryos were treated with DKK1 at Day 5 after insemination. Morulae were harvested 24 h after treatment, pooled in groups of 20 for microarray analysis and RNA subjected to analysis of gene expression by microarray hybridization. There were 662 differentially expressed genes between females and males and 128 of these genes had a fold change ≥ 1.5 between the two sexes. Of the genes upregulated in females, 49.5% were located in the X chromosome. Functional analysis predicted that cell survival was greater in female embryos. Experiment 2 involved a similar design except that transcripts for 12 genes previously reported to be affected by sex, DKK1 or the interaction were quantified by quantitative polymerase chain reaction. Expression of all genes tested that were affected by sex in experiment 1 was affected in a similar manner in Experiment 2. In contrast, effects of DKK1 on gene expression were largely not repeatable in Experiment 2. The exception was for the Hippo signaling gene AMOT, which was inhibited by DKK1. In Experiment 3, embryos produced by fertilization with unsorted sperm were treated with DKK1 at Day 5 and abundance of transcripts for CDX2, GATA6, and NANOG determined at Days 5, 6 and 7 after insemination. There was no effect of DKK1 on expression of any of the three genes. In conclusion, female and male bovine embryos have a different pattern of gene expression as early as the morula stage, and this is due to a large extent to expression of genes in the X chromosomes in females. Differential gene expression between female and male embryos is likely the basis for increased resistance to cell death signals in female embryos and disparity in responses of female and male embryos to changes in the maternal environment.