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Journal of Integrative Agriculture  2013, Vol. 12 Issue (1): 136-146    DOI: 10.1016/S2095-3119(13)60214-2
ANIMAL SCIENCE · VETERINARY SCIENCE Advanced Online Publication | Current Issue | Archive | Adv Search |
Identification of Novel and Differentially ExpressedMicroRNAs in the Ovaries of Laying and Non-Laying Ducks
 YU De-bing, JIANG Bao-chun, GONG Jing, DONG Fu-lu, LU Ying-lin, YUE Hui-jie, WANG Zhengchao
1.Department of Animal Science and Technology, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, P.R.China
2.Bioinformatics & Molecular Imaging Key Laboratory, Department of Systems Biology, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, P.R.China
3.Provincial Key Laboratory for Developmental Biology and Neurobiology, College of Life Science, Fujian Normal University, Fuzhou 350108, P.R.China
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摘要  MicroRNAs (miRNAs), which post-transcriptionally regulate gene expression by binding to the 3´ untranslated region of mRNAs to either inhibit or enhance translation, are involved in diverse biological processes. The use of high-throughput Solexa sequencing plays important roles in the discovery of miRNAs. In this study, we used high-throughput Solexa sequencing to identify novel duck miRNAs and compare the miRNA expression profiles in laying and non-laying duck ovaries. Using a bioinformatic analysis, we discovered 86 potential duck miRNAs similar to known chicken miRNAs and 43 unique sequences that matched known miRNAs of other species. We also found that miRNA variations and isoforms were widespread in our two RNA libraries, with most of the variations occurring in the 3´ region of the miRNAs. Furthermore, we detected 55 miRNAs that exhibited significant expression differences between laying and non-laying ducks. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses of the potential targets of the differentially expressed miRNAs indicated these miRNAs may play key roles in the egg laying process. Finally, we confirmed the differential expression of 5 miRNAs in the laying and non-laying samples by qRT-PCR. Cumulatively, our work provides the first look at the miRNA expression profile of the duck ovary and provides novel insight into the roles of miRNAs in egg laying and reproduction.

Abstract  MicroRNAs (miRNAs), which post-transcriptionally regulate gene expression by binding to the 3´ untranslated region of mRNAs to either inhibit or enhance translation, are involved in diverse biological processes. The use of high-throughput Solexa sequencing plays important roles in the discovery of miRNAs. In this study, we used high-throughput Solexa sequencing to identify novel duck miRNAs and compare the miRNA expression profiles in laying and non-laying duck ovaries. Using a bioinformatic analysis, we discovered 86 potential duck miRNAs similar to known chicken miRNAs and 43 unique sequences that matched known miRNAs of other species. We also found that miRNA variations and isoforms were widespread in our two RNA libraries, with most of the variations occurring in the 3´ region of the miRNAs. Furthermore, we detected 55 miRNAs that exhibited significant expression differences between laying and non-laying ducks. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analyses of the potential targets of the differentially expressed miRNAs indicated these miRNAs may play key roles in the egg laying process. Finally, we confirmed the differential expression of 5 miRNAs in the laying and non-laying samples by qRT-PCR. Cumulatively, our work provides the first look at the miRNA expression profile of the duck ovary and provides novel insight into the roles of miRNAs in egg laying and reproduction.
Keywords:  microRNA       Solexa sequencing       duck       ovary  
Received: 30 September 2011   Accepted:
Fund: 

This work was supported by the National Natural Science Foundation of China (31101705), the Natural Science Foundation of Jiangsu Province, China (BK2010454) and the Agricultural Science and Technology Independent Innovation Fund of Jiangsu Province, China (cx(09)116 and cx (11)1028).

Corresponding Authors:  Correspondence DU Wen-xing, Tel: +86-25-84395106, Fax: +86-25-84395314, E-mail: duwenxing@njau.edu.cn; GUOAn-yuan, E-mail: guoay@mail.hust.edu.cn     E-mail:  guoay@mail.hust.edu.cn
About author:  YU De-bing, E-mail: yudebing@njau.edu.cn;

Cite this article: 

YU De-bing, JIANG Bao-chun, GONG Jing, DONG Fu-lu, LU Ying-lin, YUE Hui-jie, WANG Zhengchao. 2013. Identification of Novel and Differentially ExpressedMicroRNAs in the Ovaries of Laying and Non-Laying Ducks. Journal of Integrative Agriculture, 12(1): 136-146.

[1]Bannister S C, Tizard M L, Doran T J, Sinclair A H, Smith CA. 2009. Sexually dimorphic microRNA expressionduring chicken embryonic gonadal development.Biology of Reproduction, 81, 165-176

[2]Barrell D, Dimmer E, Huntley R P, Binns D, O’Donovan C,Apweiler R. 2009. The GOA database in 2009 - anintegrated gene ontology annotation resource. NucleicAcids Research, 37, D396-D403.

[3]Bartel D P. 2004. MicroRNAs: genomics, biogenesis,mechanism, and function. Cell, 116, 281-297

[4]Betel D, Wilson M, Gabow A, Marks D S, Sander C 2008.The microRNA.org resource: targets and expression.Nucleic Acids Research, 36, D149-D153.

[5]Carletti MZ, Christenson L K. 2009. MicroRNA in the ovaryand female reproductive tract. Journal of AnimalSciences, 87, E29-E38.

[6]Chen C, Ridzon DA, BroomerAJ, Zhou Z, Lee D H, NguyenJ T, Barbisin M, Xu N L, Mahuvakar V R, Andersen MR, et al. 2005. Real-time quantification of microRNAsby stem-loop RT-PCR. Nucleic Acids Research, 33,e179.

[7]Conneely O M, Dobson A D, Carson M A, Maxwell B L,Tsai M J, SchraderWT, O’Malley B W. 1988. Structurefunctionrelationships of the chicken progesteronereceptor. Biochemical Society Transactions, 16, 683-687

[8]Elbrecht A, Lazier C B, Protter A A, Williams D L. 1984.Independent developmental programs for two estrogenregulatedgenes. Science, 225, 639-641

[9]Fiedler S D, Carletti M Z, Hong X, Christenson L K. 2008.Hormonal regulation of microRNA expression inperiovulatory mouse mural granulosa cells. Biology ofReproduction, 79, 1030-1037

[10]Friedlander M R, Chen W, Adamidi C, Maaskola J,Einspanier R, Knespel S, Rajewsky N. 2008. DiscoveringmicroRNAs from deep sequencing data using miRDeep.Nature Biotechnology, 26, 407-415

[11]George F W, Wilson J D. 1980. Endocrine differentiation ofthe fetal rabbit ovary in culture. Nature, 283, 861-863

[12]Hackl M, Jakobi T, Blom J, Doppmeier D, Brinkrolf K,Szczepanowski R, Bernhart S H, Siederdissen C H, BortJ A, Wieser M, et al. 2011. Next-generation sequencingof the Chinese hamster ovary microRNA transcriptome:Identification, annotation and profiling of microRNAsas targets for cellular engineering. Journal ofBiotechnology, 153, 62-75

[13]Hicks J A, Tembhurne PA, Liu H C. 2009. Identification ofmicroRNA in the developing chick immune organs.Immunogenetics, 61, 231-240

[14]Hicks J A, Tembhurne P, Liu H C. 2008. MicroRNAexpression in chicken embryos. Poultry Science, 87,2335-2343

[15]Hong X, Luense L J, McGinnis L K, Nothnick W B,Christenson L K. 2008. Dicer1 is essential for femalefertility and normal development of the femalereproductive system. Endocrinology, 149, 6207-6212

[16]Kanehisa M, Goto S, Kawashima S, Okuno Y, Hattori M.2004. The KEGG resource for deciphering the genome.Nucleic Acids Research, 32, D277-D280.Kim Y K, Heo I, Kim V N. 2010. Modifications of smallRNAs and their associated proteins. Cell, 143, 703-709

[17]Kozomara A, Griffiths-Jones S. 2011. MiRBase: integratingmicroRNA annotation and deep-sequencing data.Nucleic Acids Research, 39, D152-D157.

[18]Lee R C, Feinbaum R L, Ambros V. 1993. The C. elegansheterochronic gene lin-4 encodes small RNAs withantisense complementarity to lin-14

[19]Cell, 75, 843-854.Lewis B P, Burge C B, Bartel D P. 2005. Conserved seedpairing, often flanked by adenosines, indicates thatthousands of human genes are microRNA targets. Cell,120, 15-20

[20]Lin X, Liang D, He Z, Deng Q, Robertson E S, Lan K. 2011.MiR-K12-7-5p encoded by Kaposi’s sarcomaassociatedherpesvirus stabilizes the latent state bytargeting viral ORF50/RTA PLoS One, 6, e16224.

[21]Liu M, Lang N, Chen X, Tang Q, Liu S, Huang J, Zheng Y,Bi F. 2011.MiR-185 targets RhoA and Cdc42 expressionand inhibits the proliferation potential of humancolorectal cells. Cancer Letters, 301, 151-160.

[22]Lovell T M, Knight P G, Groome N P, Gladwell R T. 2001.Changes in plasma inhibin A levels during sexualmaturation in the female chicken and the effects of activeimmunization against inhibin alpha-subunit onreproductive hormone profiles and ovarian function.Biology of Reproduction, 64, 188-196

[23]Morin R D, O’Connor M D, Griffith M, Kuchenbauer F,Delaney A, Prabhu A L, Zhao Y, McDonald H, Zeng T,Hirst M, et al. 2008. Application of massively parallelsequencing to microRNA profiling and discovery inhuman embryonic stem cells. Genome Research, 18,610-621

[24]Nothnick W B. 2008. Regulation of uterine matrixmetalloproteinase-9 and the role of microRNAs Seminars in Reproductive Medicine, 26, 494-499.

[25]Novotny GW, Sonne S B, Nielsen J E, Jonstrup S P, HansenM A, Skakkebaek N E, Rajpert-De Meyts E, Kjems J,Leffers H. 2007. Translational repression of E2F1 mRNAin carcinoma in situ and normal testis correlates with expression of the miR-17-92 cluster Cell Death &Differentiation, 14, 879-882.

[26]Otsuka M, Zheng M, Hayashi M, Lee J D, Yoshino O, LinS, Han J. 2008. Impaired microRNA processing causescorpus luteum insufficiency and infertility in mice.Journal of Clinical Investigation, 118, 1944-1954.

[27]Pauley K M, Cha S. 2011. miRNA-146a in rheumatoidarthritis: a new therapeutic strategy Immunotherapy,3, 829-831.

[28]Reinhart B J, Slack F J, Basson M, Pasquinelli A E, BettingerJ C, Rougvie A E, Horvitz H R, Ruvkun G. 2000. The 21-nucleotide let-7 RNA regulates developmental timingin Caenorhabditis elegans Nature, 403, 901-906

[29]Sirotkin A V, Ovcharenko D, Grossmann R, Laukova M,Mlyncek M. 2009. Identification of microRNAscontrolling human ovarian cell steroidogenesis via agenome-scale screen. Journal of Cellular Physiology,219, 415-420

[30]Tang F, Hajkova P, Barton S C, Lao K, Surani M A. 2006.MicroRNA expression profiling of single wholeembryonic stem cells. Nucleic Acids Research, 34, e9.

[31]Wang W C, Lin F M, Chang W C, Lin K Y, Huang H D, LinN S. 2009. miRExpress: analyzing high-throughputsequencing data for profiling microRNA expression.BMC Bioinformatics, 10, 328.

[32]Ye J, Fang L, Zheng H, Zhang Y, Chen J, Zhang Z, Wang J,Li S, Li R, Bolund L. 2006.WEGO: a web tool for plottingGO annotations. Nucleic Acids Research, 34, W293-W297.
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