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雌性先熟黄鳝(Monopterus albus)卵巢分化的转录组分析

Transcriptomic analysis of the differentiating ovary of the protogynous ricefield eel Monopterus albus.

作者信息

Cai Jinfeng, Yang Wei, Chen Dong, Zhang Yize, He Zhi, Zhang Weimin, Zhang Lihong

机构信息

Department of Biology, School of Life Sciences, Sun Yat-sen University, Guangzhou, 510275, People's Republic of China.

Institute of Aquatic Economic Animals and Guangdong Province Key Laboratory for Aquatic Economic Animals, Sun Yat-Sen University, Guangzhou, 510275, P. R. China.

出版信息

BMC Genomics. 2017 Aug 3;18(1):573. doi: 10.1186/s12864-017-3953-6.

Abstract

BACKGROUND

The ricefield eel is a protogynous hermaphroditic Synbranchiform species that changes sex naturally from female to male, which offers an interesting model for studying gonadal (particularly ovarian) differentiation in vertebrates. In the present study, transcriptome sequencing of the gonad of ricefield eel larvae was performed to explore the molecular mechanisms underlying the ovarian differentiation and development.

RESULTS

A total of 301,267,988 clean reads were generated from cDNA libraries of gonadal tissues of ricefield eel larvae at 6, 9, 12, and 20 days post hatching (dph), which contained undifferentiated gonads, differentiating ovaries, ovaries with oogonia, and ovaries with meiotic oocytes, respectively. De-novo assembly of all the clean reads generated a total of 265,896 unigenes with a mean size of 720 bp and a N50 of 1107 bp. RT-qPCR analysis of the developmental expression of 13 gonadal development-related functional genes indicated that RNA-seq data are reliable. Transcriptome data suggest that high expression of female development-related genes and low expression of male development-related genes in the early gonads of ricefield eel larvae participate in the cascade of sex differentiation leading to the final female phenotype. The contrasting expression patterns of genes involved in retinoid acid (RA) synthesis and degradation might result in peak production of RA at 12 dph in the gonad of ricefield eel larvae, and induce molecular events responsible for the initiation of meiosis before the meiotic signs could be observed at 20 dph. In addition, only stra6 but not stra8 could be identified in gonadal transcriptome data of ricefield eel larvae, and the expression pattern of stra6 paralleled those of genes involved in RA synthesis, suggesting that stra6 may be a downstream target of RA and play a role in RA metabolism and/or meiotic initiation in the gonad of ricefield eel larvae.

CONCLUSIONS

The present study depicted the first large-scale RNA sequencing of the gonad of ricefield eel larvae, and identified many important functional genes, GO terms and KEGG pathways involved in gonadal development and germ cell meiosis. Results of the present study will facilitate future study on the ovarian differentiation of ricefield eels and other teleosts as well.

摘要

背景

稻田鳗鲡是一种雌雄同体的合鳃目鱼类,能自然地从雌性转变为雄性,这为研究脊椎动物性腺(尤其是卵巢)分化提供了一个有趣的模型。在本研究中,对稻田鳗鲡幼体的性腺进行转录组测序,以探索卵巢分化和发育的分子机制。

结果

从孵化后6、9、12和20天(dph)的稻田鳗鲡幼体性腺组织的cDNA文库中总共获得了301,267,988条clean reads,这些文库分别包含未分化的性腺、正在分化的卵巢、具有卵原细胞的卵巢和具有减数分裂卵母细胞的卵巢。对所有clean reads进行从头组装,共产生了265,896个单基因,平均大小为720 bp,N50为1107 bp。对13个性腺发育相关功能基因的发育表达进行RT-qPCR分析表明RNA-seq数据是可靠的。转录组数据表明,稻田鳗鲡幼体早期性腺中雌性发育相关基因的高表达和雄性发育相关基因的低表达参与了导致最终雌性表型的性别分化级联反应。参与视黄酸(RA)合成和降解的基因的对比表达模式可能导致稻田鳗鲡幼体性腺在12 dph时RA的峰值产生,并在20 dph观察到减数分裂迹象之前诱导负责减数分裂起始的分子事件。此外,在稻田鳗鲡幼体的性腺转录组数据中仅鉴定出stra6而未鉴定出stra8,并且stra6的表达模式与参与RA合成的基因的表达模式平行,这表明stra6可能是RA的下游靶点,并在稻田鳗鲡幼体性腺的RA代谢和/或减数分裂起始中发挥作用。

结论

本研究描绘了稻田鳗鲡幼体性腺的首次大规模RNA测序,并鉴定了许多参与性腺发育和生殖细胞减数分裂的重要功能基因、GO术语和KEGG途径。本研究结果也将有助于未来对稻田鳗鲡和其他硬骨鱼类卵巢分化的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7a10/5541746/65a95932bfcc/12864_2017_3953_Fig1_HTML.jpg

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