Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics

MicroRNAs (miRNAs) are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal. To discover novel miRNAs and identify the differential expression of them in ovine ovary and testis tissues, the...

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Main Authors: Wei-hua CHANG, Yong ZHANG, Zhang-rui CHENG, Xing-xu ZHAO, Juan-hong WANG, You-ji MA, Jun-jie HU, Quan-wei ZHANG
Format: Article
Language:English
Published: KeAi Communications Co., Ltd. 2015-08-01
Series:Journal of Integrative Agriculture
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Online Access:http://www.sciencedirect.com/science/article/pii/S209531191460900X
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author Wei-hua CHANG
Yong ZHANG
Zhang-rui CHENG
Xing-xu ZHAO
Juan-hong WANG
You-ji MA
Jun-jie HU
Quan-wei ZHANG
author_facet Wei-hua CHANG
Yong ZHANG
Zhang-rui CHENG
Xing-xu ZHAO
Juan-hong WANG
You-ji MA
Jun-jie HU
Quan-wei ZHANG
author_sort Wei-hua CHANG
collection DOAJ
description MicroRNAs (miRNAs) are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal. To discover novel miRNAs and identify the differential expression of them in ovine ovary and testis tissues, the study constructed two libraries by using next-generation sequencing technologies (Solexa high-throughput sequencing technique). As a result, 9 321 775 and 9 511 538 clean reads were obtained from the ovary and testis separately, which included 130 562 (90 genes of ovary) and 56 272 (85 genes of testis) of known miRNAs and 486 potential novel miRNAs reads. In this study, a total of 65 conserved miRNAs were significantly differentially expressed (P<0.01) between the two samples. Among them, 28 miRNAs were up-regulated and 3 miRNAs were down-regulated on ovary compared with testis. In addition, the known miRNAs with the highest expression level (5 miRNAs) and 30 novel miRNAs with the functions related to reproduction were validated using the real-time quantitative RT-PCR. Moreover, the gene ontology (GO) annotation and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that differentially expressed miRNAs were involved in ovary and testis physiology, including signal transduction, gonad development, sex differentiation, gematogenesis, fertilization and embryo development. The results will be helpful to facilitate studies on the regulation of miRNAs during ruminant reproduction.
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spelling doaj-art-3b56c52fe0174f299add64f486912ffd2025-08-20T03:58:22ZengKeAi Communications Co., Ltd.Journal of Integrative Agriculture2095-31192015-08-011481604161610.1016/S2095-3119(14)60900-XIdentification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformaticsWei-hua CHANG0Yong ZHANG1Zhang-rui CHENG2Xing-xu ZHAO3Juan-hong WANG4You-ji MA5Jun-jie HU6Quan-wei ZHANG7College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, P.R.China; College of Animal Science, Tarim University, Alar 843300, P.R.China; Correspondence, Mobile: +86-13893126652, Fax: +86-931-7631026College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, P.R.China; Royal Veterinary College, London University, Hatfield AL9 7TA, UK; Correspondence, Mobile: +86-13893126652, Fax: +86-931-7631026Royal Veterinary College, London University, Hatfield AL9 7TA, UKCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, P.R.ChinaCollege of Animal Science, Tarim University, Alar 843300, P.R.ChinaCollege of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, P.R.ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, P.R.ChinaCollege of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, P.R.ChinaMicroRNAs (miRNAs) are small, single stranded, non-coding RNA molecules, about 19–25 nucleotides in length, which regulate the development and functions of reproductive system of mammal. To discover novel miRNAs and identify the differential expression of them in ovine ovary and testis tissues, the study constructed two libraries by using next-generation sequencing technologies (Solexa high-throughput sequencing technique). As a result, 9 321 775 and 9 511 538 clean reads were obtained from the ovary and testis separately, which included 130 562 (90 genes of ovary) and 56 272 (85 genes of testis) of known miRNAs and 486 potential novel miRNAs reads. In this study, a total of 65 conserved miRNAs were significantly differentially expressed (P<0.01) between the two samples. Among them, 28 miRNAs were up-regulated and 3 miRNAs were down-regulated on ovary compared with testis. In addition, the known miRNAs with the highest expression level (5 miRNAs) and 30 novel miRNAs with the functions related to reproduction were validated using the real-time quantitative RT-PCR. Moreover, the gene ontology (GO) annotation and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis showed that differentially expressed miRNAs were involved in ovary and testis physiology, including signal transduction, gonad development, sex differentiation, gematogenesis, fertilization and embryo development. The results will be helpful to facilitate studies on the regulation of miRNAs during ruminant reproduction.http://www.sciencedirect.com/science/article/pii/S209531191460900XmicroRNAovarytestissolexa sequencingqRT-PCR
spellingShingle Wei-hua CHANG
Yong ZHANG
Zhang-rui CHENG
Xing-xu ZHAO
Juan-hong WANG
You-ji MA
Jun-jie HU
Quan-wei ZHANG
Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics
Journal of Integrative Agriculture
microRNA
ovary
testis
solexa sequencing
qRT-PCR
title Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics
title_full Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics
title_fullStr Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics
title_full_unstemmed Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics
title_short Identification of novel and differentially expressed microRNAs in ovine ovary and testis tissues using Solexa sequencing and bioinformatics
title_sort identification of novel and differentially expressed micrornas in ovine ovary and testis tissues using solexa sequencing and bioinformatics
topic microRNA
ovary
testis
solexa sequencing
qRT-PCR
url http://www.sciencedirect.com/science/article/pii/S209531191460900X
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