PDF2和ODF1转录组测序筛选牛卵泡发育相关基因

doi:10.11843/j.issn.0366-6964.2018.02.009

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (2): 300-309.doi: 10.11843/j.issn.0366-6964.2018.02.009

PDF2和ODF1转录组测序筛选牛卵泡发育相关基因

李鹏飞¹, 孟金柱², 郝庆玲¹, 毕锡麟³, 王锴³, 朱芷葳¹, 吕丽华^3*

1. 山西农业大学生命科学学院, 太谷 030801;
2. 铜仁学院乌江学院, 铜仁 554300;
3. 山西农业大学动物科技学院, 太谷 030801

收稿日期:2017-06-29 出版日期:2018-02-23 发布日期:2018-02-23
通讯作者: 吕丽华,E-mail:lihualvsxau@126.com
作者简介:李鹏飞(1978-),男,山西偏关人,博士,副教授,主要从事动物生殖生理方面的研究,E-mail:adamlpf@126.com
基金资助:
国家自然科学基金（31402156）；山西省回国留学人员科研资助项目（2014-重点5）；山西省国际科技合作项目（201603D421006）；山西省三晋学者和人才引进项目；山西农业大学创新基金项目（zdpy201403/201503）；引进人才博士科研启动基金（2014ZZ04）；青年拔尖创新人才支持计划（TYIT201403）；山西省重点研发计划（一般）农业项目（201703D221020-1）

Screening and Analysing of Genes Associated with Follicular Development in Bovine ODF1 and PDF2 Transcriptome

LI Peng-fei¹, MENG Jin-zhu², HAO Qing-ling¹, BI Xi-lin³, WANG Kai³, ZHU Zhi-wei¹, LÜ Li-hua^3*

1. College of Life Science, Shanxi Agricultural University, Taigu 030801, China;
2. Wujiang College, Tongren University, Tongren 554300, China;
3. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China

Received:2017-06-29 Online:2018-02-23 Published:2018-02-23
Supported by:

摘要/Abstract

摘要：

旨在筛选牛卵泡发育相关基因及其表达模式。本研究选取9头青年母牛，同期发情处理后，采集卵泡波中出现偏差前的小卵泡PDF2（The small follicle at onset of predeviation）和出现偏差后的最大卵泡ODF1（The largest follicle at onset of deviation），分离颗粒细胞（Granulesa cells，GCs），提取总RNA，并进行转录组测序；经牛RefSeq数据库搜索和差异表达基因筛选，进行GO （Gene ontology）分析；随机选择5个基因（MAPK13、CYP19A1、GREB1、SERPINE2、GSTA5）进行qRT-PCR表达量验证分析。结果表明：PDF2和ODF1转录组中共获得RPKM （The reads per kilobase per million reads）值≥0.5的表达基因15 413个，其中表达差异倍数（Fold change）≥2的差异表达基因共651个；对651个差异表达基因进行GO分析表明，参与生物过程（Biological process，BP）的基因占41.66%，分子功能（Molecular function，MF）占17.24%，细胞组分（Cellular component，CC）占41.10%；GO分析结合Genecards基因功能查询，共筛选出13个下调基因和17个上调基因与牛卵泡发育直接相关；CYP19A1、GREB1、SERPINE2在优势卵泡（Donminant follicles，DF）的表达量极显著高于从属卵泡（Subordinate follicle，SF）（P<0.01），MAPK13在SF表达量极显著高于DF （P<0.01），GSTA5在SF表达量显著高于DF （P<0.05）。5个基因差异表达趋势与转录组测序结果完全相符，也进一步证明了转录组测序结果的可信度，为深入探讨基因调控牛卵泡发育奠定了基础。

Abstract:

This study aimed to screen genes associated with bovine follicular development and investigate their expression patterns.Nine young cows were collected.The small follicle at onset of predeviation (PDF2) and the largest follicle at onset of deviation (ODF1) follicles were collected after estrus synchronization. The total RNA was extracted from separated granulosa cells for the transcriptome sequencing. The Gene Ontology analysis was carried out after searching the bovine Refseq database and screening differentially expressed genes. MAPK13,CYP19A1,GREB1,SERPINE2,GSTA5 genes were selected randomly for verification using qRT-PCR technology. Results suggested that 15 413 genes were obtained from PDF2 and ODF1 transcriptomes (RPKM value ≥ 0.5), of which 651 differentially expressed genes were selected (Fold change ≥ 2). GO assignments were used to classify the functions of the 651 genes, and which could be categorized into 3 main catrgories, biological process (41.66%), molecular function (17.24%), cellular component (41.10%). Combining with Genecards, 13 down-regulated and 17 up-regulated genes screened were associated with follicle development. qRT-PCR results indicated that the expression of CYP19A1, GREB1 and SERPINE2 in dominant follicles were very significantly greater than that in subordinate follicles (P<0.01). MAPK13 mRNA amounts in subordinate follicles were very significantly greater than that in dominant follicles (P<0.01). GSTA5 mRNA amounts in subordinate follicles were significantly greater than that in dominant follicles (P<0.05). qRT-PCR results were entirely consisted with transcriptome sequencing results, and further proved the reliability of the transcriptome sequencing, which provided a foundation for future investigation of the regulatory mechanisms involved in follicular development in cattle.

中图分类号:

S823.2

李鹏飞, 孟金柱, 郝庆玲, 毕锡麟, 王锴, 朱芷葳, 吕丽华. PDF2和ODF1转录组测序筛选牛卵泡发育相关基因[J]. 畜牧兽医学报, 2018, 49(2): 300-309.

LI Peng-fei, MENG Jin-zhu, HAO Qing-ling, BI Xi-lin, WANG Kai, ZHU Zhi-wei,Lü Li-hua. Screening and Analysing of Genes Associated with Follicular Development in Bovine ODF1 and PDF2 Transcriptome[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(2): 300-309.

0
/ / 推荐

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

链接本文: https://www.xmsyxb.com/CN/10.11843/j.issn.0366-6964.2018.02.009

https://www.xmsyxb.com/CN/Y2018/V49/I2/300

参考文献

[1] BEG M A, BERGFELT D R, KOT K, et al. Follicle selection in cattle:dynamics of follicular fluid factors during development of follicle dominance[J]. Biol Reprod, 2002, 66(1):120-126.
[2] ROMEREIM S M, SUMMERS A F, POHLMEIER W E, et al. Transcriptomes of bovine ovarian follicular and luteal cells[J]. Data Brief, 2017, 10:335-339.
[3] TERENINA E, FABRE S, BONNET A, et al. Differentially expressed genes and gene networks involved in pig ovarian follicular atresia[J]. Physiol Genomics, 2017, 49(2):67-80.
[4] LI P F, MENG J Z, LIU W Z, et al. Transcriptome analysis of bovine ovarian follicles at predeviation and onset of deviation stages of a follicular wave[J]. Int J Genomics, 2016, 2016:347248, doi:10.1155/2016/3472748.
[5] 李鹏飞, 孟金柱, 谢建山, 等. 牛卵泡ODF1与ODF2转录组发育相关基因筛选及表达差异分析[J]. 畜牧兽医学报, 2015, 46(11):1961-1966.
LI P F, MENG J Z, XIE J S, et al. Screening and analyse study of genes associated with follicular development in bovine ODF1 and ODF2 transcript[J]. Acta Veterinaria et Zootechnica Sinica, 2015, 46(11):1961-1966. (in Chinese)
[6] AUDIC S, CLAVERIE J M. The significance of digital gene expression profiles[J]. Genome Res, 1997, 7(10):986-995.
[7] XU Q, ZHAO W M, CHEN Y, et al. Transcriptome profiling of the goose (Anser cygnoides) ovaries identify laying and broodiness phenotypes[J]. PLoS One, 2013, 8(2):e55496.
[8] HATZIRODOS N, IRVING-RODGERS H F, HUMMITZSCH K, et al. Transcriptome profiling of granulosa cells of bovine ovarian follicles during growth from small to large antral sizes[J]. BMC Genomics, 2014, 15:24, doi:10.1186/1471-2164-15-24.
[9] GILBERT I, ROBERT C, DIELEMAN S, et al. Transcriptional effect of the LH surge in bovine granulosa cells during the peri-ovulation period[J]. Reproduction, 2011, 141(2):193-205.
[10] MIAO X Y, LUO Q M, QIN X Y. Genome-wide transcriptome analysis of mRNAs and microRNAs in Dorset and Small Tail Han sheep to explore the regulation of fecundity[J]. Mol Cell Endocrinol, 2015, 402:32-42.
[11] GILBERT I, ROBERT C, VIGNEAULT C, et al. Impact of the LH surge on granulosa cell transcript levels as markers of oocyte developmental competence in cattle[J]. Reproduction, 2012, 143(6):735-747.
[12] LEI M M, CAI L P, LI H, et al. Transcriptome sequencing analysis of porcine granulosa cells treated with an anti-inhibin antibody[J]. Reprod Biol, 2017, 17(1):79-88.
[13] CROUCHER N J, FOOKES M C, PERKINS T T, et al. A simple method for directional transcriptome sequencing using Illumina technology[J]. Nucleic Acids Res, 2009, 37(22):e148, doi:10.1093/nar/gkp811.
[14] O'CALLAGHAN C,FANNING L J,BARRY O P. p38δ MAPK:emerging roles of a neglected isoform[J]. Int J Cell Biol, 2014, 2014:272689, doi:10.1155/2014/272689.
[15] RISCO A, CUENDA A. New insights into the p38γ and p38δ MAPK pathways[J]. J Signal Transduct, 2012, 2012:520289, doi:10.1155/2012/520289.
[16] SUMARA G, FORMENTINI I, COLLINS S, et al. Regulation of PKD by the MAPK p38δ in insulin secretion and glucose homeostasis[J]. Cell, 2009, 136(2):235-248.
[17] OESTERREICH S. Scaffold attachment factors SAFB1 and SAFB2:innocent bystanders or critical players in breast tumorigenesis?[J]. J Cell Biochem, 2003, 90(4):653-661.
[18] IVANOVA M, DOBRZYCKA K M, JIANG S, et al. Scaffold attachment factor B1 functions in development, growth, and reproduction[J]. Mol Cell Biol, 2005, 25(8):2995-3006.
[19] TOWNSON S M,KANG K Y,LEE A V,et al.Structure-function analysis of the estrogen receptor α corepressor scaffold attachment factor-B1:identification of a potent transcriptional repression domain[J]. J Biol Chem, 2004, 279(25):26074-26081.
[20] ADAMSON E D,MERCOLA D.Egr1 transcription factor:multiple roles in prostate tumor cell growth and survival[J]. Tumour Biol, 2002, 23(2):93-102.
[21] SUKHATME V P, CAO X M, CHANG L C, et al. A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization[J].Cell, 1988, 53(1):37-43.
[22] RUSSELL D L, DOYLE K M H, GONZALES-ROBAYNA I, et al. Egr-1 induction in rat granulosa cells by follicle-stimulating hormone and luteinizing hormone:combinatorial regulation by transcription factors cyclic adenosine 3', 5'-monophosphate regulatory element binding protein, serum response factor, sp1, and early growth response factor-1[J].Mol Endocrinol, 2003, 17(4):520-533.
[23] LIM B C, MATSUMOTO S, YAMAMOTO H, et al. Prickle1 promotes focal adhesion disassembly in cooperation with the CLASP-LL5β complex in migrating cells[J]. J Cell Sci, 2016, 129(16):3115-3129.
[24] OKUDA H, MIYATA S, MORI Y, et al. Mouse Prickle1 and Prickle2 are expressed in postmitotic neurons and promote neurite outgrowth[J]. FEBS Lett, 2007, 581(24):4754-4760.
[25] TAO H, SUZUKI M, KIYONARI H, et al. Mouse prickle1, the homolog of a PCP gene, is essential for epiblast apical-basal polarity[J]. Proc Natl Acad Sci U S A, 2009, 106(34):14426-14431.
[26] WILSKER D, PROBST L, WAIN H M, et al. Nomenclature of the ARID family of DNA-binding proteins[J]. Genomics, 2005, 86(2):242-251.
[27] NISHIBUCHI G, NAKAYAMA J I. Biochemical and structural properties of heterochromatin protein 1:understanding its role in chromatin assembly[J]. J Biochem, 2014, 156(1):11-20.
[28] DOETSCHMAN T, BARNETT J V, RUNYAN R B, et al. Transforming growth factor beta signaling in adult cardiovascular diseases and repair[J]. Cell Tissue Res, 2012, 347(1):203-223.
[29] LUO W X, WILTBANK M C. Distinct regulation by steroids of messenger RNAs for FSHR and CYP19A1 in bovine granulosa cells[J]. Biol Reprod, 2006, 75(2):217-225.
[30] CAO M J, NICOLA E, PORTELA V M, et al. Regulation of serine protease inhibitor-E₂ and plasminogen activator expression and secretion by follicle stimulating hormone and growth factors in non-luteinizing bovine granulosa cells in vitro[J]. Matrix Biol, 2006, 25(6):342-354.
[31] MOHAMMED H, D'SANTOS C, SERANDOUR A A, et al. Endogenous purification reveals GREB1 as a key estrogen receptor regulatory factor[J]. Cell Rep, 2013, 3(2):342-349.

PDF2和ODF1转录组测序筛选牛卵泡发育相关基因

Screening and Analysing of Genes Associated with Follicular Development in Bovine ODF1 and PDF2 Transcriptome

RichHTML

PDF

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

编辑推荐

Metrics

本文评价

[1]	苗舒, 安济山, 王祚, 肖定福, 兰欣怡, 刘磊, 沈维军, 万发春. 亮氨酸通过PI3K-AKT信号通路促进牛成肌细胞的增殖[J]. 畜牧兽医学报, 2024, 55(1): 142-152.
[2]	牛一凡, 杨柏高, 张培培, 张航, 冯肖艺, 曹建华, 余洲, 郝海生, 杜卫华, 邹惠影, 朱化彬, 马友记, 赵学明. 牛胚胎基因组选择研究进展[J]. 畜牧兽医学报, 2023, 54(11): 4449-4457.
[3]	王文翔, 杜丽丽, 胡俊伟, 张琰翔, 马铭昊, 段蕊, 钱聪, 王欣玥, 李三禄, 张长庆, 张路培, 高雪, 徐凌洋, 李俊雅, 高会江. 基于转录组分析挖掘平凉红牛背最长肌肉质性状候选基因[J]. 畜牧兽医学报, 2023, 54(11): 4589-4604.
[4]	马浩然, 张路培, 金生云, 宝金山, 李红艳, 高会江, 徐凌洋, 王泽昭, 李俊雅. 利用高密度SNP芯片评估中国地方肉牛品种基因组亲缘关系[J]. 畜牧兽医学报, 2023, 54(10): 4174-4185.
[5]	张俊星, 张海亮, 韩丽云, 马燕芬, 温万, 周佳敏, 田佳, 路婷婷, 马云, 王雅春. 宁夏地区荷斯坦泌乳牛健康性状影响因素分析[J]. 畜牧兽医学报, 2023, 54(6): 2389-2401.
[6]	杨闯, 吴龙飞, 柳广斌, 李耀坤, 刘德武, 孙宝丽. 雷琼牛与陆丰牛背最长肌lncRNA表达特点及其相关ceRNA网络分析[J]. 畜牧兽医学报, 2023, 54(5): 1951-1963.
[7]	呙明鹏, 孟源, 王宏浩, 王元, 车雷杰, 申莉, 王曦. 晋南牛不同生长阶段体重和体尺性状遗传参数估计[J]. 畜牧兽医学报, 2023, 54(4): 1452-1464.
[8]	余诗强, 李留学, 赵小博, 赵慧颖, 屠焰, 赵玉超, 蒋林树. 不同泌乳阶段和体细胞水平的中国荷斯坦奶牛泌乳性能差异和相关性研究[J]. 畜牧兽医学报, 2023, 54(3): 1003-1014.
[9]	杨光, 徐景, 李新, 胡德宝, 郭益文, 丁向彬, 郭宏, 张林林. 干扰lncbMD对牛骨骼肌卫星细胞增殖分化的影响[J]. 畜牧兽医学报, 2023, 54(3): 1015-1025.
[10]	马志杰, 王世康, 张卫忠, 郭卫兴, 赵海明, 雷初朝. 柴达木牛和蒙古牛Y染色体基因组遗传多样性与父系起源研究[J]. 畜牧兽医学报, 2023, 54(3): 1026-1033.
[11]	牛锐利, 宋哈楠, 吴月, 王育南, 高也凡, 关伟军, 宗宪春. 西门塔尔牛表皮干细胞的分离培养与生物学特性研究[J]. 畜牧兽医学报, 2023, 54(1): 168-177.
[12]	李宏伟, 徐凌洋, 王泽昭, 蔡文涛, 朱波, 陈燕, 高雪, 张路培, 高会江, 李俊雅. 基于单倍型肉牛屠宰性状全基因组关联分析研究[J]. 畜牧兽医学报, 2022, 53(12): 4232-4243.
[13]	常瑶, 苏国生, 李艳华, 李想, 麻柱, 王雅春. 基于系谱和基因组信息估计荷斯坦青年母牛体重性状遗传参数[J]. 畜牧兽医学报, 2022, 53(11): 3759-3768.
[14]	许静漪, 徐昊祺, 胡丽蓉, 张帆, 高清, 罗汉鹏, 张海亮, 师睿, 李想, 刘林, 郭刚, 王雅春. MET基因多态性与中国荷斯坦牛繁殖和泌乳性状的关联分析[J]. 畜牧兽医学报, 2022, 53(11): 3769-3785.
[15]	王川川, 母童, 冯小芳, 禹保军, 张娟, 顾亚玲. 基于转录组测序挖掘奶牛乳脂代谢关键候选基因[J]. 畜牧兽医学报, 2022, 53(10): 3421-3433.