National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
State Key Laboratory of Livestock and Poultry Breeding, Guangdong Key Laboratory of Animal Breeding and Nutrition, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510642, China.
Int J Mol Sci. 2022 Jul 24;23(15):8157. doi: 10.3390/ijms23158157.
Litter size is an important indicator to measure the production capacity of commercial pigs. Spontaneous embryo loss is an essential factor in determining sow litter size. In early pregnancy, spontaneous embryo loss in porcine is as high as 20-30% during embryo implantation. However, the specific molecular mechanism underlying spontaneous embryo loss at the end of embryo implantation remains unknown. Therefore, we comprehensively used small RNA sequencing technology, bioinformatics analysis, and molecular experiments to determine the microRNA (miRNA) expression profile in the healthy and arresting embryo implantation site of porcine endometrium on day of gestation (DG) 28. A total of 464 miRNAs were identified in arresting endometrium (AE) and healthy endometrium (HE), and 139 differentially expressed miRNAs (DEMs) were screened. We combined the mRNA sequencing dataset from the SRA database to predict the target genes of these miRNAs. A quantitative real-time PCR assay identified the expression levels of miRNAs and mRNAs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed on differentially expressed target genes of DEMs, mainly enriched in epithelial development and amino acids metabolism-related pathways. We performed fluorescence in situ hybridization (FISH) and the dual-luciferase report gene assay to confirm miRNA and predicted target gene binding. miR-205 may inhibit its expression by combining 3'-untranslated regions (3' UTR) of tubulointerstitial nephritis antigen-like 1 (). The resulting inhibition of angiogenesis in the maternal endometrium ultimately leads to the formation of arresting embryos during the implantation period. This study provides a reference for the effect of miRNA on the successful implantation of pig embryos in early gestation.
产仔数是衡量商品猪生产能力的重要指标。胚胎自发性丢失是决定母猪产仔数的重要因素。在妊娠早期，猪胚胎着床时的自发性胚胎丢失率高达 20-30%。然而，胚胎着床末期自发性胚胎丢失的具体分子机制尚不清楚。因此，我们综合运用小 RNA 测序技术、生物信息学分析和分子实验，确定了妊娠第 28 天健康和着床中止的猪子宫内膜中的 microRNA（miRNA）表达谱。在着床中止的子宫内膜（AE）和健康子宫内膜（HE）中鉴定出 464 个 miRNA，筛选出 139 个差异表达 miRNA（DEMs）。我们结合 SRA 数据库中的 mRNA 测序数据集，预测这些 miRNA 的靶基因。通过定量实时 PCR 检测鉴定了 miRNA 和 mRNAs 的表达水平。对 DEMs 的差异表达靶基因进行了基因本体论和京都基因与基因组百科全书通路富集分析，主要富集在上皮细胞发育和氨基酸代谢相关通路。我们进行了荧光原位杂交（FISH）和双荧光素酶报告基因检测，以验证 miRNA 和预测的靶基因结合。miR-205 可能通过结合 tubulointerstitial nephritis antigen-like 1（）的 3'非翻译区（3'UTR）来抑制其表达。这导致母体子宫内膜中的血管生成受到抑制，最终导致在着床期形成着床中止的胚胎。本研究为 miRNA 对猪早期妊娠胚胎成功着床的影响提供了参考。
