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Comparative transcriptomic analysis of ovaries from high and low egg-laying Lingyun black-bone chickens.

ABSTRACT: Egg-laying rate is mainly determined by ovarian function and regulated by the hypothalamic-pituitary-gonadal axis; however, the mechanism by which the ovary regulates the egg-laying rate is still poorly understood. The purpose of this study was to compare the differences in the transcriptomes of the ovary of Lingyun black-bone chickens with relatively high and low egg-laying rates and screen candidate genes related to the egg-laying rate. RNA-sequencing (RNA-Seq) was conducted to explore the chicken transcriptome from the ovarian tissue of six Lingyun black-bone chickens with high (group G, n = 3) and low (group D, n = 3) egg-laying rates. The results showed that 235 differentially expressed genes (DEGs) were identified between the chickens with high and low egg-laying rates; among them, 209 DEGs were up-regulated and 26 DEGs were down-regulated. Gene Ontology analysis showed that the up-regulated 209 DEGs were enriched in 50 GO terms and the down-regulated 26 DEGs were enriched in 40 GO terms. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that up-regulated DEGs were significantly enriched in 25 pathways and down-regulated DEGs were significantly enriched in three pathways. Among the pathways, we found the longevity regulating pathway-multiple species pathway, Estrogen signalling pathway and PPAR signalling pathway may have an essential function in regulating the egg-laying rate. The results highlighted DEGs in the ovarian tissues of relatively high and low laying Lingyun black-bone chicken and identified essential candidate genes related to the egg-laying rate, thereby providing a theoretical basis for improving the egg-laying rate of Lingyun black-bone chicken.

SUBMITTER: Zhang Q

PROVIDER: S-EPMC8464290 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The poor egg-laying performance and short peak egg-laying period restrict the economic benefits of enterprises relating to the Taihe black-bone silky fowl. Ovaries are the main organ for egg production in poultry. Unlike that of mammals, the spawning mechanism of poultry has rarely been reported. As a prominent local breed in China, the reproductive performance of Taihe black-bone silky fowls is in urgent need of development and exploitation. To further explore the egg-laying regulation mechanism in the different periods of Taihe black-bone silky fowls, the ovarian tissues from 12 chickens were randomly selected for transcriptome analysis, and 4 chickens in each of the three periods (i.e., the pre-laying period (102 days old, Pre), peak laying period (203 days old, Peak), and late laying period (394 days old, Late)). A total of 12 gene libraries were constructed, and a total of 9897 differential expression genes (DEGs) were identified from three comparisons; the late vs. peak stage had 509 DEGs, the pre vs. late stage had 5467 DEGs, and the pre vs. peak stage had 3921 DEGs (pre-stage: pre-egg-laying period (102 days old), peak-stage: peak egg-laying period (203 days old), and late-stage: late egg-laying period (394 days old)). In each of the two comparisons, 174, 84, and 2752 differentially co-expressed genes were obtained, respectively, and 43 differentially co-expressed genes were obtained in the three comparisons. Through the analysis of the differential genes, we identified some important genes and pathways that would affect reproductive performance and ovarian development. The differential genes were LPAR3, AvBD1, SMOC1, IGFBP1, ADCY8, GDF9, PTK2B, PGR, and CD44, and the important signaling pathways included proteolysis, extracellular matrices, vascular smooth muscle contraction, the NOD-like receptor signaling pathway and the phagosome. Through the analysis of the FPKM (Fragments Per Kilobase of exon model per Million mapped fragments) values of the genes, we screened three peak egg-laying period-specific expressed genes: IHH, INHA, and CYP19A1. The twelve genes and five signaling pathways mentioned above have rarely been reported in poultry ovary studies, and our study provides a scientific basis for the improvement of the reproductive performance in Taihe black-bone silky fowls.

Project description:Introduction: Long non-coding RNA (lncRNA) refers to a category of non-coding RNA molecules exceeding 200 nucleotides in length, which exerts a regulatory role in the context of ovarian development. There is a paucity of research examining the involvement of lncRNA in the regulation of ovary development in Taihe Black-Bone Chickens. In order to further investigate the egg laying regulation mechanisms of Taihe Black-Bone Chickens at different periods, transcriptome analysis was conducted on the ovarian tissues at different laying periods. Methods: This study randomly selected ovarian tissues from 12 chickens for RNA-seq. Four chickens were selected for each period, including the early laying period (102 days, Pre), the peak laying period (203 days, Peak), and the late laying period (394 days, Late). Based on our previous study of mRNA expression profiles in the same ovarian tissue, we identified three differentially expressed lncRNAs (DE lncRNAs) at different periods and searched for their cis- and trans-target genes to draw an lncRNA-mRNA network. Results and discussion: In three groups of ovarian tissues, we identified 136 DE lncRNAs, with 8 showing specific expression during the early laying period, 10 showing specific expression during the peak laying period, and 4 showing specific expression during the late laying period. The lncRNA-mRNA network revealed 16 pairs of lncRNA-target genes associated with 7 DE lncRNAs, and these 14 target genes were involved in the regulation of reproductive traits. Furthermore, these reproductive-related target genes were primarily associated with signaling pathways related to follicle and ovary development in Taihe Black-Bone Chickens, including cytokine-cytokine receptor interaction, TGF-beta signaling pathway, tyrosine metabolism, ECM-receptor interaction, focal adhesion, neuroactive ligand-receptor interaction, and cell adhesion molecules (CAMs). This study offers valuable insights for a comprehensive understanding of the influence of lncRNAs on poultry reproductive traits.

Project description:The egg-laying rate is an important indicator for evaluating fertility of poultry. In order to better understand the laying mechanism of Muscovy ducks, gene expression profiles and pathways of ovarian tissues in high- and low-laying black (BH and BL) and white Muscovy ducks (WH and WL) during the peak production period were performed by using RNA-seq. The total number of reads produced for each ovarian sample ranged from 44,344,070 to 47,963,328. A total of 113, 619 and 87 differentially expressed genes (DEGs) were identified in BH-vs-WH, BL-vs-BH and BL-vs-WL, respectively. Among them, 54, 356 and 49 genes were up regulated and 59, 263 and 38 genes were down regulated. In addition, there were only 10 up-regulated genes in WL-vs-WH. In the comparison of DEGs in black and white Muscovy ducks, two co-expressed DEG genes were detected between BH-vs-WH and BL-vs-WL and seven DEGs were co-expressed between BL-vs-BH and WL-vs-WH. The RNA-Seq data were confirmed to be reliable by qPCR. Numerous DEGs known to be involved in ovarian development were identified, including TGFβ2, NGFR, CEBPD, CPEB2, POSTN, SMOC1, FGF18, EFNA5 and SDC4. Gene Ontology (GO) annotations indicated that DEGs related to ovarian development were mainly enriched in biological processes of "circadian sleep/wake cycle process," "negative regulation of transforming growth factor-β secretion," "positive regulation of calcium ion transport" in BH-vs-WH and "cell surface receptor signaling pathway," "Notch signaling pathway" and "calcium ion transport" in BL-vs-BH. Besides, "steroid biosynthetic process," "granulosa cell development" and "egg coat formation" were mainly enriched in BL-vs-WL and "reproduction," "MAPK cascade" and "mitotic cell cycle" were mainly enriched in WL-vs-WH. KEGG pathway analysis showed that the PI3K-Akt signaling pathway and ovarian steroidogenesis were the most enriched in Muscovy duck ovary transcriptome data. This work highlights potential genes and pathways that may affect ovarian development in Muscovy duck.

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Comparative transcriptomic analysis of ovaries from high and low egg-laying Lingyun black-bone chickens.

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