转录组测序筛选山羊妊娠早期胚胎附植相关基因

doi:10.11843/j.issn.0366-6964.2022.05.014

Abstract

Abstract: The study focused on screening key genes of embryo implantation in early pregnancy through high throughput sequencing of goat uterine horn tissue before and after embryo implantation. Sixteen multiparous female Nubian goats (Capra hircus) with similar weights((44.76±3.49) kg) and age(2-4 years old) were selected for simultaneous estrus and artificial insemination. On the 15th day (D15) and 30th day (D30) after mating, 3 goats were randomly selected and sacrificed by carotid bloodletting, respectively. The uterine horn tissues were collected after sacrifice for Illumina HiSeq high fluxtranscriptome sequencing (RNA-Seq). Differentially expressed genes (DEGs) were screened from the sequencing results, and their functions were annotated. Then, the regulatory genes related to embryo attachment were further screened through literature search. Finally, fluorescent quantitative PCR (qRT-PCR) was used to perform expression verification analysis of the selected genes. Comparative analysis of D15 and D30 uterine horn tissue transcripts obtained 2 000 DEGs, including 620 up-regulated genes and 1 380 down-regulated genes. GO functional cluster analysis was divided into 3 major categories and 52 groups, including 17 groups of cellular components, 23 groups of biological processes, and 12 groups of molecular functions, which obtained some important biological pathways such as cell connection and proliferation, biological adhesion and regulation, molecular activity and transduction, etc. Taking the expression level of genes on D15 as the standard, the gene MGP and TAGLN related to embryo attachment were screened from the 10 genes with the highest expression. From the top 10 differentially expressed genes with the largest up-regulation and down-regulation, the gene PAG-3, PAG-8 and PAG-12 related to pregnancy-associated glycoprotein synthesis and secretion, GHRHR related to growth factor binding and growth hormone releasing hormone receptor activity and IGFBP1 related to insulin-like growth factor binding protein were obtained. The results of qRT-PCR showed that the change trend of the expression of 6 randomly selected genes was consistent with the results of RNA-Seq. The obtained gene MGP, TAGLN, PAG-3, PAG-8, PAG-12, GHRHR, IGFBP1 may play an important role in the implantation and development of embryos in Nubian goat.

Key words: Nubian goat, early pregnancy, uterine horn, embryo attachment, transcriptome sequencing

CLC Number:

S827.3

LUO Jinhong, CHEN Xiang, SHANG Yishun, AO Ye, LI Pengcheng. Transcriptome Sequencing Screening the Genes Related to Goat Embryo Attachment in Early Pregnancy[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(5): 1465-1474.

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Transcriptome Sequencing Screening the Genes Related to Goat Embryo Attachment in Early Pregnancy

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