猪GV/MⅡ期卵母细胞miRNAs表达谱及Npm2基因相关miRNAs筛选

doi:10.11843/j.issn.0366-6964.2021.04.015

Abstract

Abstract: The aim of this study was to explore the miRNAs expression profile of porcine oocytes before and after in vitro maturation, and then screen miRNAs involved in regulating Npm2 gene expression. Porcine ovaries were collected at a local slaughterhouse and transported to the laboratory. GV-stage oocytes were collected and matured in vitro to obtain MⅡ-stage oocytes. Total RNA of 130 GV- or MⅡ-stage oocytes were extracted respectively and subjected to small RNA sequencing by Illumina HiSeqTM 2500. Each group was repeated 3 times. The differentially expressed miRNAs was screened, and GO and KEGG analysis were performed on the target genes. The results showed that the miRNAs expression profiles of GV- and MⅡ-stage oocytes were successfully constructed. Ninety five differentially expressed miRNAs were screened in GV- and MⅡ-stage oocytes, and miRNAs with similar expression patterns were clustered into one group. Total 3 967 target genes were obtained from 95 differentially expressed miRNAs. GO and KEGG analysis showed that these target genes were enriched in 5 194 GO terms and 212 KEGG pathways. Among these, there were two pathways related to oocyte maturation. Five miRNAs related to Npm2 gene were screened from the differentially expressed miRNAs. Two miRNAs expression were examined by qRT-PCR, and their expression trend was consistent with the sequencing results. The differentially expressed miRNAs were screened from the GV- and MⅡ-stage oocytes, it was speculated that miRNAs play a role in the process of in vitro maturation of oocytes via metabolism, oocyte meiotic related pathways and progesterone mediated oocyte maturation pathway. And miRNAs regulating Npm2 gene expression were screened. The result of this study can provide a foundation for further study the regulation mechanism of miRNAs on Npm2 gene and their roles in oocyte maturation.

Key words: pig, oocyte, miRNAs, Npm2 gene

CLC Number:

S828.3

ZHANG Ke, YAN Jimeng, LIU Yaowen, LI Honghui, CHEN Qiang, LI Zhuo, SU Yanhua, PU Shaoxia, WANG Haizhen, WEI Hongjiang, JIA Baoyu, CHENG Wenmin. miRNAs Expression Profile of Porcine GV/MⅡ-Stage Oocytes and Screening of miRNAs Related to Npm2 Gene[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(4): 996-1009.

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miRNAs Expression Profile of Porcine GV/MⅡ-Stage Oocytes and Screening of miRNAs Related to Npm2 Gene

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