影响山羊产羔数lncRNA的筛选及功能分析

doi:10.11843/j.issn.0366-6964.2020.08.009

Abstract

Abstract: The objectives of this study were to explore the correlation between lncRNAs and the reproductive regulation for goat by analyzing the expression profile of lncRNAs in ovarian tissue of Chuanzhong Black goats with high and low fecundity, and to provide the base for further interpreting the molecule mechanism of lncRNA in reproduction of goats. A total of 10 healthy Chuanzhong female goats (3.5 to 4.5 years old) with more than 3 litters were divided into high fecundity group and low fecundity group. The litter size of female goats in high fecundity group (n=6) were more than 2 per litter, the litter size of female goats in low fecundity group (n=4) were only 1 per litter. Ovary tissue of high and low fecundity goats were collected to construct the RNA sequencing libraries by extracting total RNA after estrus synchronization. Total RNA sequencing was performed by HigSeq X ten platform and differentially expressed lncRNAs were screened. Then the cis-targeted genes of lncRNAs were predicted by exploring the positional relation and binding energy between lncRNAs and mRNAs, and the GO and KEGG pathway enrichment analysis were also performed. Finally, 6 randomly selected differentially expressed lncRNAs were verified by quantitative real-time PCR. The results showed that 168 differentially expressed lncRNAs were identified. Among them, 163 lncRNAs were significantly up-regulated in goat ovaries in high fecundity group and 5 lncRNAs were significantly up-regulated in goat ovaries in low fecundity group. The screened differentially expressed lncRNAs included ENSCHIG00000001479, ENSCHIG00000002617, and ENSCHIG00000002622, which might be candidate lncRNAs for regulating fecundity of goats. The target genes of candidate lncRNAs (MYC, CDH2, FGF9, PPARGC1A, etc) were mainly involved in Janus kinase/signal transducer and activator of transcription (Jak-STAT), cell adhesion molecules, adenosine monophosphate-activated protein kinase (AMPK) signaling pathways. The target gene MYC of ENSCHIG-00000001479 participated in Jak-STAT, transforming growth factor beta (TGF-β), MAPK pathways, and the target gene CDH2 of ENSCHIG00000002617 and ENSCHIG00000002622 participated in cell adhesion molecules pathway. The results of qRT-PCR were basically consistent with the sequencing results. It was inferred that those lncRNAs(such as ENSCHIG00000001479, ENSCHIG00000002617 and ENSCHIG00000002622) might play an important role in the regulation of ovarian development and ovulation of goats. It was considered that differentially expressed lncRNAs identified were associated with litter size of goats. In this study, RNA-Seq was used to obtain and analyze differentially expressed lncRNAs in high and low fecundity goats, and GO and KEGG functional analysis was performed. This provided more transcriptome data for further exploring the reproductive mechanisms of litter size of goats.

Key words: Chuanzhong Black goats, litter size, ovary, high throughput sequencing

CLC Number:

S827.3

LI Yaokun, XU Xiangping, LU Tingting, LIAN Zhiquan, ZOU Xian, ZHAO Zhifeng, LIU Guangbin, LIU Dewu, DENG Ming. Screening and Functional Analysis of lncRNA Affecting Litter Size in Goats[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(8): 1853-1865.

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Screening and Functional Analysis of lncRNA Affecting Litter Size in Goats

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