梅山猪与杜洛克猪中等卵泡差异表达基因的鉴定

doi:10.16431/j.cnki.1671-7236.2017.02.029

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (2): 505-513.doi: 10.16431/j.cnki.1671-7236.2017.02.029

梅山猪与杜洛克猪中等卵泡差异表达基因的鉴定

赵志超^1,2, 黄涛¹, 杨飞¹, 刘丽娟¹, 孙敬礼¹, 王宵燕³, 宋成义³, 李大全¹

1. 石河子大学动物科技学院, 石河子 832003;
2. 河北工程大学生命科学与食品工程学院, 邯郸 056021;
3. 扬州大学动物科技学院, 扬州 225000

收稿日期:2016-08-18 出版日期:2017-02-20 发布日期:2017-02-25
通讯作者: 黄涛 E-mail:taohuang100@sina.com
作者简介:赵志超(1985-), 男, 安徽淮北人, 博士生, 研究方向:猪分子遗传育种, E-mail:qingxihuyang2008@163.com
基金资助:
自然科学基金（30901014、31460586）；教育部留学归国人员科研启动基金（（2013）1092）

Identification of Differentially Expressed Genes in Medium-sized Ovarian Follicles between Meishan and Duroc Sows

ZHAO Zhi-chao^1,2, HUANG Tao¹, YANG Fei¹, LIU Li-juan¹, SUN Jing-li¹, WANG Xiao-yan³, SONG Cheng-yi³, LI Da-quan¹

1. College of Animal Science and Technology, Shihezi University, Shihezi 832003, China;
2. School of Life Science and Food Engineering, Hebei University of Engineering, Handan 056021, China;
3. College of Animal Science and Technology, Yangzhou University, Yangzhou 225000, China

Received:2016-08-18 Online:2017-02-20 Published:2017-02-25

摘要/Abstract

摘要：

本试验采用抑制性消减杂交技术构建了卵泡期第4天梅山猪和杜洛克猪中等卵泡M2组织差异表达的消减cDNA文库，从中筛选差异表达的基因，并通过实时荧光定量PCR对其进行验证，利用DAVID软件对差异表达的基因进行聚类分析。结果表明，从梅山猪和杜洛克猪M2卵泡消减文库中筛选得到了148和75个差异表达的ESTs，分别含有125和60个已知的基因。实时荧光定量PCR验证结果与筛选结果相符。GO功能分类注释到调控代谢、细胞循环、生物合成、胞内转运、类固醇雌激素受体和刺激等生物学过程。KEGG Pathway分析表明有6个基因参与TGF-beta信号通路，5个基因参与卵母细胞减数分裂信号通路，7个基因参与类固醇雌激素受体信号通路，推测TGF-beta信号通路中的基因可能调控猪卵泡的发育。研究结果为深入探讨卵泡发育机理和对繁殖性状影响的遗传机理奠定了基础。

关键词: 差异表达基因; 卵泡; 梅山猪; 杜洛克猪

Abstract:

To reveal the molecular mechanism involved in different number of ovulation between hyperprolific and ordinary sows, forward and reverse subtracted cDNA libraries were constructed to screen differentially expressed genes in medium ovarian follicles at 4th day during follicular phase of estrous cycle between Meishan and Duroc sows. The differentially expressed genes selected were demonstrated by Real-time PCR and cluster analysis was performed through DAVID. The results showed that a total of 148 and 75 non-redundant ESTs were isolated from the SSH library of Meishan and Duroc sows M2 follicles, including 125 and 60 known genes, respectively. The results of Real-time PCR were consistent with the screening results. GO analysis indicated that these genes were involved in regulation of metabolic process, cell cycle, biosynthetic process, intracellular transport, steroid hormone receptor and stimulus. KEGG pathway analysis defined 6 genes involved in TGF-beta signaling pathway, 5 genes in oocyte meiosis pathway, 7 genes in steroid hormone receptor signaling pathway. Genes controlling TGF-beta signaling pathway were considered to play an important role in regulating follicle development. The research would be helpful for further studying the follicular development and the genetic mechanism on reproductive traits.

Key words: differentially expressed genes; ovarian follicle; Meishan sow; Duroc sow

中图分类号:

S814.8

赵志超, 黄涛, 杨飞, 刘丽娟, 孙敬礼, 王宵燕, 宋成义, 李大全. 梅山猪与杜洛克猪中等卵泡差异表达基因的鉴定[J]. 《中国畜牧兽医》, 2017, 44(2): 505-513.

ZHAO Zhi-chao, HUANG Tao, YANG Fei, LIU Li-juan, SUN Jing-li, WANG Xiao-yan, SONG Cheng-yi, LI Da-quan. Identification of Differentially Expressed Genes in Medium-sized Ovarian Follicles between Meishan and Duroc Sows[J]. , 2017, 44(2): 505-513.

参考文献

[1] Miller A T, Picton H M, Craigon J, et al. Follicle dynamics and aromatase activity in high-ovulating Meishan sows and in Large-White hybrid contemporaries[J]. Biol Reprod,1998, 58(6):1372-1378.
[2] Yen H W, Ford J, Zimmerman D, et al. Follicular development and maturation in gilts selected for an index of high ovulation rate and high prenatal survival[J]. J Anim Sci, 2005, 83(1):130-135.
[3] Wolter B F, Ellis M, Curtis S E, et al. Group size and floor-space allowance can affect weanling-pig performance[J]. J Anim Sci, 2000, 78(8): 2062-2067.
[4] 杨公社.猪生产学[M].北京:中国农业出版社,2002.
[5] Bazer F, Thatcher W, Martinat-Botte F, et al. Sexual maturation and morphological development of the reproductive tract in Large White and prolific Chinese Meishan pigs[J]. J Reprod Fertil, 1988, 83(2):723-728.
[6] Bolet G, Botte F M, Locatelli A, et al. Components of prolificacy in hyperprolific Large White sows compared with the Meishan and Large White breeds[J]. Genet Sel Evol, 1986,18(3):333-342.
[7] 李炳坦,赵书广,郭传甲.养猪生产技术手册[M].第2版.北京:中国农业出版社, 2004.
[8] Davis K L, Robison O W,Toelle V D. Breed differences in uterine and ovarian measurements in gestating swine[J]. J Anim Sci,1987, 65(3):685-691.
[9] Foxcroft G R, Hunter M G. Basic physiology of follicular maturation in the pig[J]. J Reprod FertilSuppl,1985,33:1-19.
[10] Yen H W. Follicular development, maturation and atresia during the estrous cycle in gilts expressing high and low ovulation rates[D].Lincoln:University of Nebraska,1999.
[11] Yen H W, Ford J J, Zimmerman D R,et al. Follicular development and maturation in gilts selected for an index of high ovulation rate and high prenatal survival[J]. J Anim Sci, 2005, 83(1):130-135.
[12] Grado-Ahuir J A, Aad P Y, Ranzenigo G, et al. Microarray analysis of insulin-like growth factor-Ⅰ-induced changes in messenger ribonucleic acid expression in cultured porcine granulose cells: Possible role of insulin-like growth factor-Ⅰ inangiogenesis[J]. J Anim Sci, 2009, 87(6):1921-1933.
[13] Paradis F, Moore H S, Pasternak J A, et al. Pig preovulatory oocytes modulate cumulus cell protein and gene expression in vitro[J]. Mol Cell Endocrinol,2010, 320(1-2): 87-96.
[14] Toms D, Tsoi S, Dobrinsky J, et al. The effects of glial cell line-derived neurotrophic factor on the in vitro matured porcine oocyte transcriptome[J]. Mol Reprod Dev,2014, 81(3): 217-229.
[15] Shi Y, Massagué J. Mechanisms of TGF-β signaling from cell membrane to the nucleus[J]. Cell, 2003, 113(6):685-700.
[16] Onichtchouk D, Chen Y G, Dosch R, et al. Silencing of TGF-β signalling by the pseudoreceptor BAMBI[J]. Nature, 1999,401(6752):480-485.
[17] 杨飞,黄涛,赵志超,等. 杜洛克与梅山猪大卵泡消减文库的构建[J]. 石河子大学学报, 2014, 32(5):543-549.
[18] Sun X, Mei S, Tao H, et al. Microarray profiling for differential gene expression in PMSG-hCG stimulated preovulatory ovarian follicles of Chinese Taihu and Large White sows[J]. BMC Genomics, 2011,12:111-120.
[19] Zhang Q, Sun H, Jiang Y, et al. microRNA-181a suppresses mouse granulosa cell proliferation by targeting activin receptor ⅡA[J]. PLoS One, 2013, 8(3):e59667.
[20] Li Q, Pangas S A, Jorgez C J, et al. Redundant roles of SMAD2 and SMAD3 in ovarian granulosa cells in vivo[J]. Mol Cell Biol, 2008, 28(23):7001-7011.
[21] 徐梦思,黄涛,刘丽娟,等.猪SKP1基因的克隆及其在梅山猪与杜洛克猪卵泡中的表达研究[J]. 中国畜牧兽医,2015, 42(7):1640-1646.
[22] De S, Kline D. Evidence for the requirement of 14-3-3eta (YWHAH) in meiotic spindle assembly during mouse oocyte maturation[J]. BMC Dev Biol, 2013, 13:1-15.
[23] De S, Marcinkiewicz J L, Vijayaraghavan S, et al. Expression of 14-3-3 protein isoforms in mouse oocytes, eggs and ovarian follicular development[J]. BMC Research Notes, 2012, 5:57.
[24] 刘丽娟,鲁慧文,杨飞,等.YWHAG基因CDS的克隆及其在梅山与杜洛克猪卵泡发育中的差异表达研究[J]. 畜牧兽医学报,2014, 45(3):489-493.
[25] Lanzino M, De Amicis F, McPhaul M, et al. Endogenous coactivator ARA70 interacts with estrogen receptor alpha (ERalpha) and modulates the functional ERalpha/androgen receptor interplay in MCF-7 cells[J]. J Biol Chem, 2005, 280(21):20421-20430.
[26] Gao T, Brantley K, Bolu E, et al. RFG (ARA70, ELE1) interacts with the human androgen receptor in a ligand-dependent fashion, but functions only weakly as a coactivator in cotransfection assays[J]. Molecular Endocrinology, 1999, 13(10):1645-1656.
[27] Ting H J, Bao B Y, Hsu C L, et al. Androgen-receptor coregulators mediate the suppressive effect of androgen signals on vitamin D receptor activity[J]. Endocrine, 2005, 26(1):1-9.
[28] Moore J M, Galicia S J, McReynolds A C, et al. Quantitative proteomics of the thyroid hormone receptor-coregulator interactions[J]. J Biol Chem, 2004, 279(26):27584-27590.
[29] Viswakarma N, Jia Y, Bai L, et al.Coactivators in PPAR-regulated gene expression[J]. PPAR Research, 2010, 2010:250126.

梅山猪与杜洛克猪中等卵泡差异表达基因的鉴定

Identification of Differentially Expressed Genes in Medium-sized Ovarian Follicles between Meishan and Duroc Sows

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