山羊卵巢富集chi-miR-100f-5p的鉴定与表达分析

doi:10.3864/j.issn.0578-1752.2013.22.016

中国农业科学 ›› 2013, Vol. 46 ›› Issue (22): 4784-4790.doi: 10.3864/j.issn.0578-1752.2013.22.016

• 畜牧·兽医·资源昆虫 • 上一篇下一篇

山羊卵巢富集chi-miR-100f-5p的鉴定与表达分析

娜日苏12, 刘广1, 向阳1, 赵海林1, 吴亭1, 赵金红2, 李凡1, 黄选洋1, 孙晓卫1, 郑姣1, 张家骅1

1.西南大学动物科技学院/重庆市牧草与草食家畜重点实验室/重庆市草食动物资源保护与利用工程技术研究中心，重庆 400715
2.重庆市畜牧科学院，重庆 400716

收稿日期:2013-01-25 出版日期:2013-11-15 发布日期:2013-04-23
通讯作者: 通信作者张家骅，Tel：023-68250215；E-mail：jhzhang007@yahoo.com.cn
作者简介:娜日苏，Tel：023-68251196；E-mail：narisu193@sina.com。刘广，Tel：023-68251196；E-mail：jianliuxue@sina.com
基金资助:
中央高校基本科研业务费专项资金（XDJK2010C101）、西南大学博士基金项目（SWU111048）、国家自然科学基金项目（31172195）、重庆市科技攻关计划项目（CSTC，2011AB1084）、重庆市畜牧科学院基本科研业务项目（10604）

Characterization and Expression Analysis of Goat Ovarian chi-miR-100f-5p

NA Ri-Su-12, LIU Guang-1, XIANG Yang-1, ZHAO Hai-Lin-1, WU Ting-1, ZHAO Jin-Hong-2, LI Fan-1, HUANG Xuan-Yang-1, SUN Xiao-Wei-1, ZHENG Jiao-1, ZHANG Jia-Hua-1

1.College of Animal Science and Technology, Southwest University/Chongqing Key Laboratory of Forage and Herbivore/Chongqing Engineering Research Center for Herbivores Resource Protection and Utilization, Chongqing 400715
2.Chongqing Academy of Animal Husbandry, Chongqing 400716

Received:2013-01-25 Online:2013-11-15 Published:2013-04-23

摘要/Abstract

摘要： 【目的】探讨山羊卵巢中富集的miR-100及预测的靶基因与山羊卵巢功能及卵泡发育的潜在关系，为山羊繁殖机理研究提供数据。【方法】以山羊卵巢组织为材料，通过miRNA的分离、克隆及测序得到山羊卵巢中表达的miR-100，采用RT-qPCR进行表达检测，用生物信息学方法预测miR-100的靶基因及其功能。【结果】① 山羊卵巢的测序结果中，一段成熟体为21nt 的序列与果蝇dan-miR-100高度同源，将其命名为chi-miR-100f-5p,其前体位于基因组负（-）链上，长度为59 bp，定位于山羊CM000899.1:33 997 894—33 997 914序列；② chi-miR-100f-5p 在山羊卵巢、垂体及心脏等组织中均有表达，说明此miRNA不是卵巢组织所特有；心脏和垂体组织中均有较高表达，其次是卵巢，而在肝脏、脾脏、肺及肾脏中表达显著低于卵巢（P ＜0.01）；在单产羔内蒙古绒山羊卵巢中chi-miR-100f-5p的表达极显著高于高繁殖力大足黑山羊卵巢（P＜0.01）；③ chi-miR-100f-5p主要靶作用于整合蛋白β1亚基基因及垂体特异性转录因子1基因，可能参与整合蛋白信号通路，促进垂体内分泌细胞的生长与分化，调节下丘脑-垂体-性腺轴激素分泌。【结论】山羊卵巢中新发现的chi-miR-100f-5p 可能是与繁殖调控有关的重要候选miRNAs 之一。

关键词: chi-miR-100f-5p , 微小RNA , 卵巢

Abstract: 【Objective】 The relationship between the goat ovarian expressed miR-100 with its predicted target genes and goat ovarian function or follicular development was studied in order to provide data for goat breeding mechanism.【Method】 Goat ovarian expressed miR-100 was obtained by cloning and sequencing a small RNA library with pooled RNA from goat ovarian tissue. The expression profiling was detected by RT-qPCR. miR-100 target genes and functional classification were predicted by bioinformatics method.【Result】The chi-miR-100f-5p, at a length of 21 nt , was isolated from goat ovaries. The sequence is highly homologous with dan-miR-100, and has one loci in the genome negative (-) chain, which located in goats sequence CM000899.1: 33997894 to 33997914; The chi-miR-100f-5p was detected in goat ovarian, pituitary, heart and other tissues, showing chi-miR- 100f-5p was not an ovarian-specific miRNA. It was found that chi-miR-100f-5p expressed at higher level in heart and pituitary, secondly in ovary, and the expression of chi-miR-100f-5p in the liver, spleen, lung, and kidney was significantly lower than in ovary (P＜0.01). It was alsofound that chi-miR-100f-5p expressed at significantly higher level in single lambing Inner Mongolia cashmere goat ovary than the high fecundity Dazu black goat ovary (P＜0.01); The chi-miR-100f-5p was mainly targeted for integrin β1 subunit and pituitary-specific transcription factor 1 gene, and might be involved in the integrin signaling pathway, promoting the growth and differentiation of pituitary endocrine cells, and regulating the hypothalamic-pituitary-gonadal axis hormonessecretion.【Conclusion】The newly discovered chi-miR-100f-5p in goat ovarian may be an important candidate miRNA in regulation of breeding.

Key words: chi-miR-100f-5p , microRNA , ovary

娜日苏12, 刘广1, 向阳1, 赵海林1, 吴亭1, 赵金红2, 李凡1, 黄选洋1, 孙晓卫1, 郑姣1, 张家骅1. 山羊卵巢富集chi-miR-100f-5p的鉴定与表达分析[J]. 中国农业科学, 2013, 46(22): 4784-4790.

NA Ri-Su-12, LIU Guang-1, XIANG Yang-1, ZHAO Hai-Lin-1, WU Ting-1, ZHAO Jin-Hong-2, LI Fan-1, HUANG Xuan-Yang-1, SUN Xiao-Wei-1, ZHENG Jiao-1, ZHANG Jia-Hua-1. Characterization and Expression Analysis of Goat Ovarian chi-miR-100f-5p[J]. Scientia Agricultura Sinica, 2013, 46(22): 4784-4790.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2013.22.016

https://www.chinaagrisci.com/CN/Y2013/V46/I22/4784

参考文献

[1]Carletti M Z, Christenson L K. MicroRNA in the ovary and female reproductive tract. Journal of Animal Science, 2009, 87(Suppl.4): E29-E38.

[2]Christenson L K. MicroRNA control of ovarian function. Animal Reproduction, 2010, 7(3): 129-133.

[3]O’Hara S P, Mott J L, Splinter P L. MicroRNAs: key modulators of posttranscriptional gene expression. Gastroenterology, 2009, 136: 17-25.

[4]Ramesh S P. MicroRNA function: multiple mechanisms for a tiny RNA. RNA, 2005, 11(12): 1753-1761.

[5]Bartel D P, Chen C Z. Micromanagers of gene expression: the potentially widespread influence of metazoan microRNAs. Nature Reviews Genetics, 2004, 5 (5): 396-400.

[6]Kim H J, Cui X S, Kim E J, Kin W J, Kim N H. New porcine microRNA genes found by homology search. Genome, 2006, 49(10): 1283-1286.

[7]Ro S, Song R, Park C, Zheng H, Sanders K M, Yan W. Cloning and expression profiling of small RNAs expressed in the mouse ovary. RNA, 2007, 13(12): 2366-2380.

[8]Hossain M M, Ghanem N, Hoelker M, Rings F, Phatsara C, Tholen E, Schellander K, Tesfaye D. Identification and characterization of miRNAs expressed in the bovine ovary. BMC Genomics, 2009, 10: 443.

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

[10]Lei L, Jin S, Gonzale G, Behringer R R, Woodruff T K. The regulatory role of Dicer in folliculogenesis in mice. Molecular and Cellular Endocrinology, 2009, 315(1/2): 63-73.

[11]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.

[12]Yao N, Yang B Q, Liu Y, Tan X Y, Lu C L, Yuan X H, Ma X. Follicle-stimulating hormone regulation of microRNA expression on progesterone production in cultured rat granulosa cells. Endocrine, 2010, 38(2): 158-166.

[13]Fiedler S D, Carletti M Z, Hong X M, Christenson L K. Hormonal regulation of MicroRNA expression in periovulatory mouse mural granulosa cells. Biology of Reproduction, 2008, 79(6): 1030-1037.

[14]Yao G D, Yin M M, Lian J, Tian H, Liu L, Li X, Sun F. MicroRNA-224 is involved in transforming growth factor-beta- mediated mouse granulosa cell proliferation and granulosa cell function by targeting Smad 4. Molecular Endocrinology, 2010, 24(3): 540-551.

[15]Nagaraja A K, Creighton C J, Yu Z F, Zhu H F, Gunaratne P H, Reid J G, Olokpa E, Itamochi H, Ueno N T, Hawkins S M, Anderson M L, Matzuk M M. A link between mir-100 and FRAP1/mTOR in clear cell ovarian cancer. Molecular Endocrinology, 2010, 24(2): 447-463.

[16]Gebeshuber C A, Martinez J. MiR-100 suppresses IGF2 and inhibits breast tumorigenesis by interfering with proliferation and survival signaling. Oncogene, 2012, 10: 1038.

[17]Peng D X, Luo M, Qiu L W, He Y L, Wang X F. Prognostic implications of microRNA-100 and its functional roles in human epithelial ovarian cancer. Oncology Reports, 2012, 27(4): 1238-1244.

[18]Sebastian G D, Felix P H, Sheena K. MicroRNA-100 regulates neovascularization by suppression of mammalian target of rapamycin in endothelial and vascular smooth muscle cells. Circulation, 2011, 123(9): 999-1009.

[19]Carolina T, Gaetana P, Luca C, Giuseppina M, Lucio P, Silvia P, Tommaso R. MiRNA 34a, 100, and 137 modulate differentiation of mouse embryonic stem cells. The FASEB Journal Research Communication, 2009, 24(9): 3255-3263.

[20]Mourelatos Z, Dostie J, Paushkin S, Sharma A, Charroux B, Abel L, Rappsilber J, Mann M, Dreyfuss G. MiRNPs: a novel class of ribonucleoproteins containing numerous microRNAs. Genes Development, 2002, 16(6): 720-728.

[21]Yang H, Kong W, He L, Zhao J J, O’Donnell J D, Wang J, Wenham R M, Coppola D, Kruk P A, Nicosia S V, Cheng J Q. MicroRNA expression profiling in human ovarian cancer: miR-214 induces cell survival and cisplatin resistance by targeting PTEN. Cancer Research, 2008, 68(2): 425- 433.

[22]Nam E J, Yoon H, Kim S W, Kim H, Kim Y T, Kim J H, Kim J W, Kim S. MicroRNA expression profiles in serous ovarian carcinoma. Clinical Cancer Research, 2008, 14: 2690 -2695.

[23]Sucharov C, Bristow M R, Port J D. MiRNA expression in the failing human heart: functional correlates. Journal of Molecular and Cellular Cardiology, 2008, 45(2): 185-192.

[24]Henson B J, Bhattacharjee S, O’Dee D M, Feingold E, Gollin S M. Decreased expression of miR-125b and miR-100 in oral cancer cells contributes to malignancy. Genes Chromosomes Cancer, 2009, 48(7): 569-582.

[25]陈翠, 张立敏, 陈晓杰, 刘喜冬. 垂体特异性转录因子１基因遗传多态性与中国西门塔尔牛生长及育肥性状的遗传效应分析. 中国畜牧兽医, 2012, 39(4): 122-125.

Chen C, Zhang L M, Chen X J, Liu X D. Analysis the effects of the polymorphism of pituitary specific transcription factor 1 gene on growth and fattening characters of Chinese Simmental. China Animal Husbandry and Veterinary Medicine, 2012, 39(4): 122-125. (in Chinese)

[26]韩毅冰, 蔡志明, 孙青原, 陈系古. 整合蛋白β1 在人类卵母细胞和早期胚胎上分布的研究. 解剖学报, 2002, 33(4): 400-403.

Han Y B, Cai Z M, Sun Q Y, Chen X G. Study of the distribution of integrin β1 on human oocytes and early embryos. Chinese Journal of Anatomy, 2002, 33(4): 400-403. (in Chinese)

山羊卵巢富集chi-miR-100f-5p的鉴定与表达分析

Characterization and Expression Analysis of Goat Ovarian chi-miR-100f-5p

PDF

赞

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0

[1]	刘玉芳,陈玉林,周祖阳,储明星. miR-221-3p靶向BCL2L11调控小尾寒羊卵泡颗粒细胞凋亡[J]. 中国农业科学, 2022, 55(9): 1868-1876.
[2]	阿依木古丽·阿不都热依木,阿尔祖古丽·阿依丁,王家敏,石嘉琛,马芳芳,蔡勇,乔自林. 大豆异黄酮对牦牛卵巢颗粒细胞增殖和凋亡的影响[J]. 中国农业科学, 2022, 55(8): 1667-1675.
[3]	李丽莹,何颖婷,钟玉宜,周小枫,张豪,袁晓龙,李加琪,陈赞谋. CTNNB1对猪卵巢颗粒细胞的调控[J]. 中国农业科学, 2022, 55(15): 3050-3061.
[4]	张静,张姬越,岳永起,赵丹,范依琳,马妍,熊燕,熊显荣,字向东,李键,杨丽雪. LKB1基因对卵巢颗粒细胞类固醇激素生成相关基因的调控作用[J]. 中国农业科学, 2022, 55(10): 2057-2066.
[5]	冯睿蓉,付中民,杜宇,张文德,范小雪,王海朋,万洁琦,周紫彧,康育欣,陈大福,郭睿,史培颖. 中华蜜蜂幼虫肠道中微小RNA的鉴定及分析[J]. 中国农业科学, 2022, 55(1): 208-218.
[6]	杜宇,范小雪,蒋海宾,王杰,冯睿蓉,张文德,余岢骏,隆琦,蔡宗兵,熊翠玲,郑燕珍,陈大福,付中民,徐国钧,郭睿. 微小RNA介导意大利蜜蜂工蜂对东方蜜蜂微孢子虫的跨界调控[J]. 中国农业科学, 2021, 54(8): 1805-1820.
[7]	杜星,曾强,刘禄,李琦琦,杨柳,潘增祥,李齐发. 二花脸猪linc-NORFA核心启动子鉴定与转录调控分析[J]. 中国农业科学, 2021, 54(15): 3331-3342.
[8]	张鑫,霍孔林,宋星星,张多妮,胡文,胡传活,李珣. GnIH通过p38MAPK信号通路对猪卵巢颗粒细胞自噬与凋亡的影响[J]. 中国农业科学, 2020, 53(9): 1904-1912.
[9]	陈华枝,祝智威,蒋海宾,王杰,范元婵,范小雪,万洁琦,卢家轩,熊翠玲,郑燕珍,付中民,陈大福,郭睿. 蜜蜂球囊菌菌丝和孢子中微小RNA及其靶mRNA的比较分析[J]. 中国农业科学, 2020, 53(17): 3606-3619.
[10]	耿四海,石彩云,范小雪,王杰,祝智威,蒋海宾,范元婵,陈华枝,杜宇,王心蕊,熊翠玲,郑燕珍,付中民,陈大福,郭睿. 微小RNA介导东方蜜蜂微孢子虫侵染意大利蜜蜂工蜂的分子机制[J]. 中国农业科学, 2020, 53(15): 3187-3204.
[11]	杜宇,范小雪,蒋海宾,王杰,范元婵,祝智威,周丁丁,万洁琦,卢家轩,熊翠玲,郑燕珍,陈大福,郭睿. 微小RNA及其介导的竞争性内源RNA调控网络在意大利蜜蜂工蜂中肠发育过程中的潜在作用[J]. 中国农业科学, 2020, 53(12): 2512-2526.
[12]	辛晓萍, 王家迎, 张爱玲, 钟玉宜, 何颖婷, 陈赞谋, 张哲, 张豪, 李加琪, 袁晓龙. 转录因子CEBPα和p53在猪卵巢颗粒细胞中对Kiss1基因表达的调控[J]. 中国农业科学, 2019, 52(9): 1624-1634.
[13]	李文杨,刘远,吴贤锋,高承芳,黄勤楼. 福清山羊与努比亚黑山羊发情期卵巢组织RNA-Seq分析[J]. 中国农业科学, 2019, 52(12): 2171-2182.
[14]	郭睿,杜宇,童新宇,熊翠玲,郑燕珍,徐国钧,王海朋,耿四海,周丁丁,郭意龙,吴素珍,陈大福. 意大利蜜蜂幼虫肠道在球囊菌侵染前期的差异表达microRNA及其调控网络[J]. 中国农业科学, 2019, 52(1): 166-180.
[15]	赵静, 陶蓉, 郝德君, 肖留斌, 谭永安. 甜菜夜蛾卵黄原蛋白受体基因的RNA干扰[J]. 中国农业科学, 2019, 52(1): 56-64.