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高通量测序分析确定了与鸭卵巢产蛋相关的微小RNA。

High-throughput sequencing analysis identified microRNAs associated with egg production in ducks ovaries.

作者信息

Qiu Mohan, Zhang Zengrong, Xiong Xia, Du Huarui, Li Qingyun, Yu Chunlin, Gan Wu, Liu Hehe, Peng Han, Xia Bo, Chen Jialei, Hu Chenming, Song Xiaoyan, Yang Li, Jiang Xiaosong, Yang Chaowu

机构信息

Sichuan Animal Science Academy, Chengdu, China.

Animal Breeding and Genetics Key Laboratory of Sichuan Province, Chengdu, China.

出版信息

PeerJ. 2020 Feb 4;8:e8440. doi: 10.7717/peerj.8440. eCollection 2020.

Abstract

BACKGROUND

MicroRNAs (miRNAs) exist widely and are involved in multiple biological processes in ducks, whereas the regulatory mechanism of miRNAs in egg laying of ducks has remained unclear. This study aims to reveal key miRNAs involved in the regulation of egg production in duck ovaries.

METHODS

High-throughput sequencing was performed on four egg-type duck ovaries and four egg-meat-type duck ovaries at the start of the egg-laying stage. Quantitative reverse transcription PCR (qRT-PCR) validation was performed on differentially expressed miRNAs (DE miRNAs). Gene network of DEmiRNA-mRNA-pathway was constructed by Cytoscape.

RESULTS

A total of 251 know miRNAs and 1,972 novel miRNAs were obtained from whole clean reads. Among the known miRNAs, we identified 21 DEmiRNAs, including eight down-regulated and 13 up-regulated miRNAs in egg-type ducks compared with egg-meat-type ducks. Among the novel miRNAs, we identified 70 DEmiRNAs, including 58 down-regulated and 12 up-regulated in egg-type ducks compared with egg-meat-type ducks. The expression patterns of four miRNAs were verified by qRT-PCR. The DEmiRNAs were involved in the function of response to folic acid and the pathway of valine, leucine and isoleucine degradation. Specific target genes of DEmiRNAs enrichment was found in some egg-laying regulation pathways, such as dopaminergic synapse, ovarian steroidogenesis and oocyte meiosis. The DEmiRNA-mRNA-pathway network including three DEmiRNAs, nine mRNAs and 11 pathways. apl-miR-194-5p and apl-miR-215-5p may be potential key miRNAs in regulating egg laying.

CONCLUSIONS

This study provided miRNAs profiles in ducks about egg laying and establish a theoretical basis for subsequent selection or modification of duck phenotypes at the molecular level.

摘要

背景

微小RNA(miRNA)广泛存在,参与鸭的多种生物学过程,而miRNA在鸭产蛋中的调控机制尚不清楚。本研究旨在揭示参与鸭卵巢产蛋调控的关键miRNA。

方法

在产蛋期开始时,对四个蛋用型鸭卵巢和四个蛋肉兼用型鸭卵巢进行高通量测序。对差异表达的miRNA(DE miRNA)进行定量逆转录PCR(qRT-PCR)验证。利用Cytoscape构建DEmiRNA-mRNA-通路基因网络。

结果

从全部清洁读段中获得了251个已知miRNA和1972个新miRNA。在已知miRNA中,我们鉴定出21个DEmiRNA,与蛋肉兼用型鸭相比,蛋用型鸭中有8个下调miRNA和13个上调miRNA。在新miRNA中,我们鉴定出70个DEmiRNA,与蛋肉兼用型鸭相比,蛋用型鸭中有58个下调miRNA和12个上调miRNA。通过qRT-PCR验证了四个miRNA的表达模式。这些DEmiRNA参与了对叶酸的应答功能以及缬氨酸、亮氨酸和异亮氨酸降解途径。在一些产蛋调控途径中发现了DEmiRNA富集的特定靶基因,如多巴胺能突触、卵巢类固醇生成和卵母细胞减数分裂。DEmiRNA-mRNA-通路网络包括三个DEmiRNA、九个mRNA和11条通路。apl-miR-194-5p和apl-miR-215-5p可能是调控产蛋的潜在关键miRNA。

结论

本研究提供了鸭产蛋的miRNA图谱,为后续在分子水平上选择或改良鸭的表型奠定了理论基础。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/74d3/7006514/739a30d5ea84/peerj-08-8440-g001.jpg

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