伊犁鹅产蛋前后各期卵巢转录谱的构建及卵泡发育相关基因的分析

doi:10.16431/j.cnki.1671-7236.2022.12.017

Abstract

Abstract: 【Objective】 The experiment was aimed to construct a transcriptome library of Yili goose ovaries before and after egg laying, combined with bioinformatics analysis, reveal differentially expressed genes in ovarian tissues of Yili geese at different egg laying stages, identify key genes affecting goose ovarian development, and provide theoretical references for reproductive regulation of Yili geese.【Method】 Four Yili geese at the prelaying period(KL), laying period(CL), and ceased period(XL) were selected, and the ovarian tissues were obtained after slaughter for the construction of ovaries transcriptome libraries.Candidate genes associated with follicle development were screened by novel gene mining, differential gene expression, gene annotation, and protein interaction network analysis.Eight differentially expressed genes were randomly selected and Real-time quantitative PCR was used to verify their expression.【Result】 Histological results of the ovaries of Yili geese showed that a large number of primary follicles on the surface of the ovaries at the prelaying period, while the ovaries at the laying period showed a hierarchy of follicles, and follicles at the ceased period could be observed to appear sunken and atretic inwards.By RNA sequencing (RNA-Seq), 57 811 186 to 85 328 377 clean reads were obtained from the 12 constructed Yili goose ovary cDNA libraries, all with Q30 values were greater than 93.38%, and the sequencing reads produced by each sample were compared to the goose reference genome at a rate of 82.79% to 89.24%.A total of 1 112 novel genes were annotated in ovaries, and the total number of single nucleotide polymorphism loci (SNP) was ranged from 1 642 273 to 2 425 069.SNP and InDel were mainly annotated in the intron region.The alternative splicing in each period were mainly concentrated in TSS and TTS.There were 337, 1 136 and 525 differentially expressed genes in KL vs CL, XL vs CL and XL vs KL groups, respectively, and the total differentially expressed genes were adrenoceptor alpha 2A (ADRA2A), cuticular protein (CP), glycoprotein non-metastatic melanoma protein B (GPNMB) and alpha-1-antitrypsin-like (LOC106033756).GO function enrichment analysis revealed that the differential mRNAs were mainly enriched in positive regulation of peptidyl-tyrosine phosphorylation, cell adhesion and external side of plasma membrane.KEGG pathway enrichment analysis revealed that differentially expressed genes were significantly enriched mainly in neuroactive ligand-receptor interaction, ECM-receptor interaction and steroid biosynthesis.Combined with protein interaction network analysis, potential regulatory factors Bruton’s tyrosine kinase (BTK), platelet derived growth factor receptor alpha (PDGFRA), integrin subunit beta 3 (ITGB3) associated with goose ovarian development were screened.The results of Real-time quantitative PCR showed that RNA-Seq results were accurate and reliable.【Conclusion】 This study revealed the gene expression differences in ovarian tissues of Yili geese before and after egg laying, and screened out important pathways (neuroactive ligand-receptor interaction, ECM-receptor interaction, steroid biosynthesis) and key candidate genes (BTK, PDGFRA and ITGB3), which could provide theoretical support for understanding the molecular mechanism of ovarian tissues in regulating egg laying performance of Yili geese.

Key words: Yili geese; egg laying cycle; ovary development; transcriptome

CLC Number:

S835

ZHAO Xiaoyu, WU Yingping, LI Haiying, YAO Yingying, LI Jiahui, YAO Yang, LU Qingqing. Construction of Ovarian Transcriptional Profiles and Analysis of Follicle Development-Related Genes in Yili Geese at Various Stages Before and After Egg Laying[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4674-4687.

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Construction of Ovarian Transcriptional Profiles and Analysis of Follicle Development-Related Genes in Yili Geese at Various Stages Before and After Egg Laying

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