应用Solexa测序技术挖掘山羊卵巢组织microRNA

doi:10.3864/j.issn.0578-1752.2013.01.017

摘要/Abstract

摘要： 【目的】旨在构建、分析山羊卵巢组织miRNA表达谱，为进一步研究特定miRNA与山羊卵泡发育及性激素分泌等相关生殖活动的关系奠定理论基础。【方法】首先从山羊卵巢组织总RNA中分离小片段RNA，然后进行Solexa测序和生物信息学分析，最后利用q-PCR技术验证miRNA在山羊卵巢组织中的表达。【结果】共鉴定出508个山羊卵巢组织表达的miRNA和19个对应的miRNA*，这些miRNA在哺乳动物（绵羊、牛、猪、马、狗）进化过程中高度保守；q-PCR验证了8个miRNA在山羊卵巢组织中的表情况，定量结果与测序结果一致。【结论】成功构建了山羊卵巢组织miRNA表达文库，山羊卵巢组织miRNA表达丰富且其表达量各异。

关键词: miRNA , Solexa , 卵巢 , 安徽白山羊

Abstract: 【Objective】 Characterization of microRNA expression in hircine ovary tissues was studied, with the purposes of providing basic information for further studies of relationships between specific miRNAs and hircine reproduction such as folliculogenesis and sex hormone secretion. 【Method】 The small RNAs isolated from total RNA of ovary tissues were sequenced by Solexa and then bioinformatics analysis were perfomed. Meanwhile, the expression of selected miRNAs was validated by q-PCR. 【Result】Sequence analysis indicated that 508 ovary miRNAs and 19 corresponding miRNA*s were identified which are conservative in evolution of mammals (sheep, bovine, swine, horse, dog). The expression of 8 selected miRNAs in ovary tissues obtained by q-PCR was in agreement with Solexa sequencing results. 【Conclusion】 The expression profiling of miRNAs, which are abundant and differentially expressed in hircine ovary tissues, was constructed successfully.

Key words: miRNA , Solexa , ovary , Anhui White goat

张晓东, 凌英会, 张运海, 李运生, 刘亚, 曹鸿国, 张子军, 殷宗俊, 丁建平, 章孝荣. 应用Solexa测序技术挖掘山羊卵巢组织microRNA[J]. 中国农业科学, 2013, 46(1): 146-153.

ZHANG Xiao-Dong, LING Ying-Hui, ZHANG Yun-Hai, LI Yun-Sheng, LIU Ya, CAO Hong-Guo, ZHANG Zi-Jun, YIN Zong-Jun, DING Jian-Ping, ZHANG Xiao-Rong. MicroRNAs in Ovaries of Goats (Capra hircus) Identified by Solexa Sequencing[J]. Scientia Agricultura Sinica, 2013, 46(1): 146-153.

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参考文献

[1]Ambros V. The functions of animal microRNAs. Nature, 2004, 431(7006): 350-355.

[2]Esau C, Kang X, Peralta E, Hanson E, Marcusson E G, Ravichandran L V, Sun Y, Koo S, Perera R J, Jain R, Dean N M, Freier S M, Bennett C F, Lollo B, Griffey R. MicroRNA-143 regulates adipocyte differentiation. The Journal of Biological Chemistry, 2004, 279(50): 52361-52365.

[3]Inoue K. MicroRNA function in animal development. FEBS Letter, 2005, 579(26): 5911-5912.

[4]Baley J, Li J. MicroRNAs and ovarian function. Journal of Ovarian Research, 2012, 5: 8-14.

[5]郭婧, 董旻岳. 微小RNA和生殖. 国际生殖健康/计划生育杂志, 2010, 29(1): 11-14.

Guo J, Dong M Y. MicroRNA and reproduction. Journal of Infernational Reproductive Health/Family Planning, 2010, 29(1): 11-14. (in Chinese)

[6]俞焙秦, 刘炳亚. miRNA的生物学特性和功能. 上海交通大学学报: 医学版, 2007, 27(5): 621-623.

Yu B Q, Liu B Y. Biological character and function of miRNA. Journal of Shanghai Jiaotong University: Medical Science, 2007, 27(5): 621-623. (in Chinese)

[7]Cutting A D, Bannister S C, Doran T J, Sinclair A H, Tizard M V, Smith C A. The potential role of microRNAs in regulating gonadal sex differentiation in the chicken embryo. Chromosome Research, 2012, 20(1): 201-213.

[8]Otsuka M, Zheng M, Hayashi M, Lee J D, Yoshino O, Lin S, Han J. Impaired microRNA processing causes corpus luteum insufficiency and infertility in mice. The Journal of Clinical Investigation, 2008, 118(5): 1944-1954.

[9]Ahn H W, Morin R D, Zhao H, Harris R A, Coarfa C, Chen Z J, Milosavljevic A, Marra M A, Rajkovic A. MicroRNA transcriptome in the newborn mouse ovaries determined by massive parallel sequencing. Molecular Human Reproduction, 2010, 16(7): 463-471.

[10]Huang J, Ju Z, Li Q, Hou Q, Wang C, Li J, Li R, Wang L, Sun T, Hang S, Gao Y, Hou M, Zhong J. Solexa sequencing of novel and differentially expressed microRNAs in testicular and ovarian tissues in Holstein cattle. International Journal of Biological Sciences, 2011, 7(7): 1016-1026.

[11]Deo A, Carlsson J, Lindlof A. How to choose a normalization strategy for miRNA quantitative real-time (qPCR) arrays. Journal of Bioinformatics and Computational Biology, 2011, 9(6): 795-812.

[12]朱芷葳, 贺俊平, 程志学, 王海东, 李鹏飞, 乔德瑞, 董常生. microRNA在成年羊驼皮肤组织中的表达. 畜牧兽医学报, 2010, 41(10): 1342-1345.

Zhu Z W, He J P, Cheng Z X, Wang H D, Li P F, Qiao D R, Dong C S. microRNA Expression in adut alpaca skin. Acta Veterinaria et Zootechnica Sinica, 2010, 41(10): 1342-1345. (in Chinese)

[13]习欠云, 周莲莲, 李虹仪, 韩东, 束刚, 王松波, 高萍, 朱晓彤, 王修启, 江青艳, 张永亮. 不同品种猪肌肉组织miR-1和miR-133基因的表达分析. 畜牧兽医学报, 2012, 43(6): 843-848.

Xi Q Y, Zhou L L, Li H Y, Han D, Shu G, Wang S B, Gao P, Zhu X T, Wang X Q, Jiang Q Y, Zhang Y L. The expression analysis of miR-1 and miR-133 in longissimus dorsi muscle from different pig breeds. Acta Veterinaria et Zootechnica Sinica, 2012, 43(6): 843-848. (in Chinese)

[14]Amoah E A, Gelaye S. Biotechnological advances in goat reproduction. Journal of Animal Science, 1997, 75(2): 578-585.

[15]Zhang C Y, Chen S L, Li X, Xu D Q, Zhang Y, Yang L G. Genetic and phenotypic parameter estimates for reproduction traits in the Boer dam. Livestock Science, 2009, 125(1): 60-65.

[16]Leboeuf B, Delgadillo J A, Manfredi E, Piacere A, Clement V, Martin P, Pellicer M, Boue P, de Cremoux R. Management of goat reproduction and insemination for genetic improvement in France. Reproduction in Domestic Animals, 2008, 43(Suppl. 2): 379-385.

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

[18]Wei Y, Chen S, Yang P, Ma Z, Kang L. Characterization and comparative profiling of the small RNA transcriptomes in two phases of locust. Genome Biology, 2009, 10(1): R6.

[19]Fu Y, Shi Z, Wu M, Zhang J, Jia L, Chen X. Identification and differential expression of microRNAs during metamorphosis of the Japanese flounder (Paralichthys olivaceus). PloS One, 2011, 6(7): e22957.

[20]Hsieh M, Johnson M A, Greenberg N M, Richards J S. Regulated expression of Wnts and frizzleds at specific stages of follicular development in the rodent ovary. Endocrinology, 2002, 143(3): 898-908.

[21]Ricken A, Lochhead P, Kontogiannea M, Farookhi R. Wnt signaling in the ovary: identification and compartmentalized expression of wnt-2, wnt-2b, and frizzled-4 mRNAs. Endocrinology, 2002, 143(7): 2741-2749.

[22]Takanabe R, Ono K, Abe Y, Takaya T, Horie T, Wada H, Kita T, Satoh N, Shimatsu A, Hasegawa K. Up-regulated expression of microRNA-143 in association with obesity in adipose tissue of mice fed high-fat diet. Biochemical and Biophysical Research Communications, 2008, 376(4): 728-732.

[23]Wang T, Li M, Guan J, Li P, Wang H, Guo Y, Shuai S, Li X. MicroRNAs miR-27a and miR-143 regulate porcine adipocyte lipid metabolism. International Journal of Molecular Sciences, 2011, 12(11): 7950-7959.

[24]Yi C, Xie W D, Li F, Lü Q, He J, Wu J, Gu D, Xu N, Zhang Y. MiR-143 enhances adipogenic differentiation of 3T3-L1 cells through targeting the coding region of mouse pleiotrophin. FEBS Letters, 2011, 585(20): 3303-3309.

[25]Carletti M Z, Fiedler S D, Christenson L K. MicroRNA 21 blocks apoptosis in mouse periovulatory granulosa cells. Biology of Reproduction, 2010, 83(2): 286-295.

[26]Fu T Y, Lin C T, Tang P C. Steroid hormone-regulated let-7b mediates cell proliferation and basigin expression in the mouse endometrium. The Journal of Reproduction and Development, 2011, 57(5): 627-635.

[27]Saleh A D, Savage J E, Cao L, Soule B P, Ly D, DeGraff W, Harris C C, Mitchell J B, Simone N L. Cellular stress induced alterations in microRNA let-7a and let-7b expression are dependent on p53. PloS One, 2011, 6(10): e24429.

[28]Roush S, Slack F J. The let-7 family of microRNAs. Trends in Cell Biology, 2008, 18(10): 505-516.

[29]Miles J R, McDaneld T G, Wiedmann R T, Cushman R A, Echternkamp S E, Vallet J L, Smith T P. MicroRNA expression profile in bovine cumulus-oocyte complexes: possible role of let-7 and miR-106a in the development of bovine oocytes. Animal Reproduction Science, 2012, 130(1/2): 16-26.