Maillo Veronica, Gaora Peadar Ó, Forde Niamh, Besenfelder Urban, Havlicek Vitezslav, Burns Gregory W, Spencer Thomas E, Gutierrez-Adan Alfonso, Lonergan Patrick, Rizos Dimitrios
Departamento de Reproducción Animal, INIA, Madrid, Spain.
School of Biomolecular and Biomedical Sciences, University College Dublin, Belfield, Dublin, Ireland.
Biol Reprod. 2015 Jun;92(6):144. doi: 10.1095/biolreprod.115.127969. Epub 2015 Apr 29.
This study examined the effect of the presence of single or multiple embryos on the transcriptome of the bovine oviduct. In experiment 1, cyclic (nonbred, n = 6) and pregnant (artificially inseminated, n = 11) heifers were slaughtered on Day 3 after estrus, and the ampulla and isthmic regions of the oviduct ipsilateral to the corpus luteum were separately flushed. Oviductal epithelial cells from the isthmus region, in which all oocytes/embryos were located, were snap-frozen for microarray analysis. In experiment 2, heifers were divided into cyclic (nonbred, n = 6) or pregnant (multiple embryo transfer, n = 10) groups. In vitro-produced presumptive zygotes were transferred endoscopically to the ipsilateral oviduct on Day 1.5 postestrus (n = 50 zygotes/heifer). Heifers were slaughtered on Day 3, and oviductal isthmus epithelial cells were recovered for RNA sequencing. Microarray analysis in experiment 1 failed to detect any difference in the transcriptome of the oviductal isthmus induced by the presence of a single embryo. In experiment 2, following multiple embryo transfer, RNA sequencing revealed 278 differentially expressed genes, of which 123 were up-regulated and 155 were down-regulated in pregnant heifers. Most of the down-regulated genes were related to immune function. In conclusion, the presence of multiple embryos in the oviduct resulted in the detection of differentially expressed genes in the oviductal isthmus; failure to detect changes in the oviduct transcriptome in the presence of a single embryo may be due to the effect being local and undetectable under the conditions of this study.
本研究考察了单个或多个胚胎的存在对牛输卵管转录组的影响。在实验1中,发情周期母牛(未配种,n = 6)和妊娠母牛(人工授精,n = 11)在发情后第3天屠宰,黄体同侧输卵管的壶腹部和峡部区域分别冲洗。峡部区域中所有卵母细胞/胚胎所在部位的输卵管上皮细胞迅速冷冻用于微阵列分析。在实验2中,母牛被分为发情周期组(未配种,n = 6)或妊娠组(多个胚胎移植,n = 10)。在发情后第1.5天,将体外产生的推定受精卵经内镜转移至同侧输卵管(每头母牛50个受精卵)。母牛在第3天屠宰,回收输卵管峡部上皮细胞用于RNA测序。实验1中的微阵列分析未能检测到单个胚胎的存在对输卵管峡部转录组产生的任何差异。在实验2中,多个胚胎移植后,RNA测序显示有278个差异表达基因,其中123个在妊娠母牛中上调,155个下调。大多数下调基因与免疫功能相关。总之,输卵管中多个胚胎的存在导致了输卵管峡部差异表达基因的检测;在单个胚胎存在的情况下未能检测到输卵管转录组的变化,可能是由于这种影响具有局部性且在本研究条件下无法检测到。