绵羊季节性发情性状核心基因和关键lncRNA的筛选与分析

doi:10.11843/j.issn.0366-6964.2025.03.026

Abstract

Abstract:

The aim of this study was to screen core genes and long non-coding RNAs related to seasonal estrus. Ovarian tissue were collected from 2 to 3 years old, non-pregnant female Chinese Merino and Hu sheep (3 sheep per group, each in anestrus, estrus and interestrus). RNA-seq technology was employed to identify differentially expressed genes and lncRNAs. GO and KEGG enrichment analyses, protein-protein interaction (PPI) network analysis and ceRNA network construction were performed to identify core genes and lncRNAs related to reproductive performance. RNA libraries for 6 distinct physiological states were constructed. There were 1 495 differentially expressed genes (DEGs) and 454 differentially expressed long non-coding RNAs (lncRNAs) be identified. Among these, 313 DEGs and 435 lncRNAs were common, while 1 182 DEGs and 19 lncRNAs were unique. GO and KEGG enrichment analyses revealed that common DEGs were significantly enriched in the processes related to cell division and the cell cycle, whereas the target genes of differentially expressed lncRNAs were enriched in intracellular signaling pathways and negative regulation of RNA polymerase Ⅱ promoter transcription. Unique DEGs were primarily associated with the positive regulation of cell migration. The target genes of lncRNAs were enriched in cell adhesion and integrin-mediated signaling pathways. There were 19 core genes identified by protein-protein interaction network analysis, which were predominantly enriched in oocyte meiosis and cell cycle pathways. These core genes exhibited higher expression level during the anestrus phase than those in interestrus. Further analysis of the ceRNA network highlighted the enrichment of genes in cell cycle-related signaling pathways. RT-qPCR results were consisted with the RNA-seq results. A total of 19 core genes and 28 lncRNAs were identified, along with 42 lncRNA-miRNA-mRNA interactions, providing potential regulatory targets for understanding the molecular mechanisms of seasonal reproduction in sheep.

Key words: sheep, seasonal estrus, RNA-seq, core gene, lncRNA

CLC Number:

S826.3

YANG Yang, LI Liangyuan, WAN Pengcheng, LU Shouliang, LIU Changbin, YANG Hua, WANG Limin, DAI Rong, ZHOU Ping. Screening and Analysis of Core Genes and Key lncRNAs for Seasonal Estrus Traits in Sheep[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1264-1277.

Figures/Tables 13

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基因 Gene	序列(5'→3') Primer sequence	产物大小/bp Product size	参考序列号 Accession number
A2M	F: CGGGCACAGTTCTCCAT R: GGCAAGGGCTCAAGGTA	185	ENSOARG00000000950
E114	F: TGTGAGACCGTCAAATGG R: CAGAACAAAGGCTGTAAGG	144	ENSOARG00000000114
FSHR	F: TGACACCAAGATAGCCAAGCG R: GGTAGAACAGGACCAGGAGGA	149	NM_001009289.1
GAPDH	F: CTGACCTGCCGCCTGGAGAAA R: TAGAAGAGTGAGTGTCGCTGTT	149	NM_001190390.1

品种 Breed	样品 Sample	质控后序列 Clean reads	匹配/reads Mapped on reference	GC/%	Q30/%
中国美利奴羊 Chinese Merino sheep	M_A	105 030 108	100 750 493(95.93%)	46.66	92.99
	MBE	87 990 548	83 968 497(95.43%)	54.39	92.91
	MBD	83 665 146	78 978 206(94.40%)	54.83	92.86
湖羊 Hu sheep	HBE	87 506 822	83 480 433(95.40%)	53.15	93.09
	HAE	90 475 066	86 278 630(95.36%)	45.40	92.94
	HAD	76 988 138	73 770 028(95.82%)	49.78	93.08

分组 Group	lncRNA		DEGs
分组 Group	up	down	up	down
M_A vs. MBD	235	231	1 396	744
M_A vs. MBE	209	254	1 510	600
M_A vs. HAD	209	266	1 876	1 206
M_A vs. HAE	236	235	556	537
M_A vs. HBE	233	240	1 400	774

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Screening and Analysis of Core Genes and Key lncRNAs for Seasonal Estrus Traits in Sheep

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