牦牛发情期卵巢比较转录组学研究

doi:10.11843/j.issn.0366-6964.2016.09.011

摘要/Abstract

摘要：

旨在进一步了解牦牛发情期卵巢的分子机制，解析牦牛繁殖的特殊性。本研究应用RNA-seq技术对牦牛和平原黄牛发情期卵巢进行转录组高通量测序及全基因组差异表达模式比对分析。通过比较分析牦牛和黄牛卵巢转录组数据，共筛选出1 307个差异表达基因，其中661个基因表达量上调和646个基因表达量下调。进一步功能分析表明，这些差异基因涉及多种 GO分类及KEGG通路。其中GO分类注释显示，差异基因与细胞粘附、激素调控等生物学过程存在密切关联，同时钙离子结合、阳离子跨膜转运等分子事件表现活跃。KEGG通路分析显示，补体和凝血级联通路的富集水平最高，其次为细胞色素P450相关通路。昼夜节律等一些新型通路，尽管与生殖功能没有明显关联，但也表现出显著富集。本研究首次对比牦牛和黄牛发情期卵巢转录组数据，筛选并分析相关差异基因。该研究结果为进一步阐述牦牛卵巢的基本分子机理提供基础，同时也为全面理解牦牛繁殖特异性提供新的思路。

关键词: 牦牛, 发情期卵巢, 转录组, 比较分析, 分子机制

Abstract:

For elaborate the general physiological molecular features of the estrus ovary of yaks and particularity of yak reproduction.RNA-seq technology was applied to analyze transcriptome data comparatively between the yak and plain cattle estrus ovaries in this study.Compared with cattle，genome-wide divergent expression patterns during the ovary transcriptome data revealed that 1 307 genes were significantly and differentially expressed，in which 661 genes were upregulated and 646 genes were downregulated.Functional analysis showed that the differentially expressed genes were involved in various GO categories and KEGG pathways.GO annotations indicated that the genes were closely related to cell adhesion，hormonal biological processes，whereas the calcium ion binding，cation transmembrane transport molecular events were significantly active.KEGG pathway analysis showed that the complement and coagulation cascade pathway was the most enriched，followed by the cytochrome P450 related pathways.Moreover，several novel pathways，such as circadian rhythm，were significantly enriched despite having no evident associations with the reproductive function.This study is the first to compare the ovary transcriptome data between yak and cattle and to identify and analyze the related differential genes.Our findings provide a theory support for further investigation of the molecular mechanism of yak ovary and offer new insights to understand comprehensively the specificity of yak reproduction.

Key words: yak, estrus ovary, transcriptome, comparative analysis, general molecular mechanism

中图分类号:

S823.8+ 5.2

兰道亮，熊显荣，柴志欣，艾鷖，黄偲，李键. 牦牛发情期卵巢比较转录组学研究[J]. 畜牧兽医学报, 2016, 47(9): 1830-1839.

LAN Dao-liang，XIONG Xian-rong，CHAI Zhi-xin，AI Yi，HUANG Cai，LI Jian. Comparative Transcriptome Analysis between Yak and Cattle Estrus Ovary[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(9): 1830-1839.

参考文献

［1］QIU Q，ZHANG G，MA T，et al.The yak genome and adaptation to life at high altitude［J］.Nat Genet，2012，44(8)：946-949.
［2］XIONG X，FU M，LAN D，et al.Yak response to high-altitude hypoxic stress by altering mRNA expression and DNA methylation of hypoxia-inducible factors［J］.Anim Biotechnol，2015，26(3)：222-229.
［3］MIAO F，GUO Z，XUE R，et al.Effects of grazing and precipitation on herbage biomass，herbage nutritive value，and yak performance in an alpine meadow on the Qinghai-Tibetan Plateau［J］.PLoS One，2015，10(6)：e0127275.
［4］FU M，CHEN Y，XIONG X，et al.Establishment of mammary gland model in vitro：culture and evaluation of a yak mammary epithelial cell line［J］.PLoS One，2014，9(12)：e113669.
［5］GUO X，LONG R，KREUZER M，et al.Importance of functional ingredients in yak milk-derived food on health of Tibetan nomads living under high-altitude stress：a review［J］.Crit Rev Food Sci Nutr，2014，54(3)：292-302.
［6］LONG R J，ZHANG D G，WANG X，et al.Effect of strategic feed supplementation on productive and reproductive performance in yak cows［J］. Prev Vet Med，1999，38 (3)：195-206.
［7］PRAKASH B S，SARKAR M，MONDAL M.An update on reproduction in yak and mithun［J］.Reprod Domest Anim，2008，43（Suppl 2）：217-223.
［8］FU M，XIONG X R，LAN D L，et al.Molecular characterization and tissue distribution of estrogen receptor genes in domestic yak［J］.Asian-Australas J Anim Sci，2014，27(12)：1684-1690.
［9］XIAO X，ZI X D，NIU H R，et al.Effect of addition of FSH，LH and proteasome inhibitor MG132 to in vitro maturation medium on the developmental competence of yak (Bos grunniens) oocytes［J］.Reprod Biol Endocrinol，2014，12：30.
［10］PAN L，GONG W，ZHOU Y，et al.A comprehensive transcriptomic analysis of infant and adult mouse ovary［J］.Genomics Proteomics Bioinformatics，2014，12(5)：239-248.
［11］MIAO X，LUO Q.Genome-wide transcriptome analysis between small-tail Han sheep and the Surabaya fur sheep using high-throughput RNA sequencing［J］.Reproduction，2013，145(6)：587-596.
［12］CUI Y，YU S J.An anatomical study of the internal genital organs of the yak at different ages［J］.Vet J，1999，157(2)：192-196.
［13］BECKER J，HACKL M，RUPP O，et al.Unraveling the Chinese hamster ovary cell line transcriptome by next-generation sequencing［J］. J Biotechnol，2011，156(3)：227-235.
［14］权凯，张兆旺.母牦牛的繁殖特性［J］.畜牧与饲料科学，2005，26(5)：30-32.
QUAN K，ZHANG Z W.Reproductive characteristics of female yak［J］.Animal Husbandry and Food Science，2005，26(5)：30-32.(in Chinese)
［15］ZI X D，HE S M，LU H，et al.Induction of estrus in suckled female yaks (Bos grunniens) and synchronization of ovulation in the non-sucklers for timed artificial insemination using progesterone treatments and Co-Synch regimens［J］.Anim Reprod Sci，2006，92(2)：183-192.
［16］HU Q，MA T，WANG K，et al.The yak genome database：an integrative database for studying yak biology and high-altitude adaption［J］.BMC Genomics，2012，13：600.
［17］AUDIC S，CLAVERIE J M.The significance of digital gene expression profiles［J］.Genome Res，1997，7(10)：986-995.
［18］EPPIG J J.Intercommunication between mammalian oocytes and companion somatic cells［J］.Bioessays，1991，13(11)：569-574.
［19］GILCHRIST R B，RITTER L J，ARMSTRONG D T.Oocyte-somatic cell interactions during follicle development in mammals［J］.Anim Reprod Sci，2004，83：431-446.
［20］MAKKI M，SABOORI E，SABBAGHI M A，et al.Effects of selenium，calcium and calcium ionophore on human oocytes in vitro maturation in a chemically defined medium［J］.Iran J Reprod Med，2012，10(4)：343-348.
［21］ROZINEK J，RAJMON R，PETR J，et al.Ultrastructural localisation of calcium deposits in pig ovarian follicles［J］.Anim Reprod Sci，2006，91(2)：123-132.
［22］HEDIGER M A.Membrane permeability.The diversity of transmembrane transport processes［J］.Curr Opin Cell Biol，1997，9(4)：543-546.
［23］KIM A M，BERNHARDT M L，KONG B Y，et al.Zinc sparks are triggered by fertilization and facilitate cell cycle resumption in mammalian eggs［J］.ACS Chem Biol，2011，6(7)：716-723.
［24］ANCHORDOQUY J M，ANCHORDOQUY J P，SIRINI M A，et al.The importance of having zinc during in vitro maturation of cattle cumulus-oocyte complex：role of cumulus cells［J］.Reprod Domest Anim，2014，49(5)：865-874.
［25］AZUMA T，KONDO T，IKEDA S，et al.Effects of EDTA saturated with Ca2+ (Ca-EDTA) on pig，bovine and mouse oocytes at the germinal vesicle stage during maturation culture and the involvement of chelation of Zn2+ in pronuclear formation induction by Ca-EDTA［J］.Reproduction，2002，124(2)：235-240.
［26］SUNYER J O，BOSHRA H，LORENZO G，et al.Evolution of complement as an effector system in innate and adaptive immunity［J］.Immunol Res，2003，27(3)：549-564.
［27］APPLEDORN D M，MCBRIDE A，SEREGIN S，et al.Complex interactions with several arms of the complement system dictate innate and humoral immunity to adenoviral vectors［J］.Gene Ther，2008，15(24)：1606-1617.
［28］MOISEYEV G，GIVLI S，BAR-YOSEPH P Z.Fibrin polymerization in blood coagulation-a statistical model［J］.J Biomech，2013，46(1)：26-30.
［29］HARRIS C L，MIZUNO M，MORGAN B P.Complement and complement regulators in the male reproductive system［J］.Mol Immunol，2006，43(2)：57-67.
［30］MASTELLOS D，GERMENIS A E，LAMBRIS J D.Complement：an inflammatory pathway fulfilling multiple roles at the interface of innate immunity and development［J］.Curr Drug Targets Inflamm Allergy，2005，4(1)：125-127.
［31］HASTY L A，LAMBRIS J D，LESSEY B A，et al.Hormonal regulation of complement components and receptors throughout the menstrual cycle［J］.Am J Obstet Gynecol，1994，170(1)：168-175.
［32］DANIELSON P B.The cytochrome P450 superfamily：biochemistry，evolution and drug metabolism in humans［J］.Curr Drug Metab，2002，3(6)：561-597.
［33］KANDIEL M M，WATANABE G，TAYA K.Ovarian expression of inhibin-subunits，3beta-hydroxysteroid dehydrogenase，and cytochrome P450 aromatase during the estrous cycle and pregnancy of Shiba goats (Capra hircus)［J］.Exp Anim，2010，59(5)：605-614.
［34］SAHMI M，NICOLA E S，PRICE C A.Hormonal regulation of cytochrome P450 aromatase mRNA stability in non-luteinizing bovine granulosa cells in vitro［J］.J Endocrinol，2006，190(1)：107-115.
［35］FRADETTE C，DU SOUICH P.Hypoxia-inducible factor-1 and activator protein-1 modulate the upregulation of CYP3A6 induced by hypoxia［J］.Br J Pharmacol，2003，140(6)：1146-1154.
［36］SIMONSON T S，YANG Y，HUFF C D，et al.Genetic evidence for high-altitude adaptation in Tibet［J］.Science，2010，329(5987)：72-75.
［37］EDERY I.Circadian rhythms in a nutshell［J］. Physiol Genomics，2000，3(2)：59-74.
［38］PIGGINS H D.Human clock genes［J］.Ann Med，2002，34(5)：394-400.

[1]	王璟, 滑留帅, 陈俊峰, 张家庆, 任巧玲, 白红杰, 郭红霞, 徐照学, 邢宝松, 白献晓, 曹海. 去势对淮南公猪背最长肌转录组的影响[J]. 畜牧兽医学报, 2019, 50(9): 1746-1758.
[2]	荣敏, 张然然, 涂剑峰, 王天娇, 孙丽敏, 杨颖, 徐佳萍, 邢秀梅*. 美国短毛黑水貂快速生长过程中转录组差异表达分析[J]. 畜牧兽医学报, 2019, 50(9): 1787-1801.
[3]	韩小红, 何翃闳, 王靖雷, 张慧珠, 马悦, 马进彪, 赵生贤, 余四九, 崔燕, 樊江峰. 牦牛p38MAPK在雌性主要生殖器官中的表达[J]. 畜牧兽医学报, 2019, 50(9): 1802-1812.
[4]	赵学亮, 王姝懿, 孙柯, 苏倩, 王文龙, 刘春霞. 捻转血矛线虫阿苯达唑敏感株和耐药株比较转录组学分析[J]. 畜牧兽医学报, 2019, 50(9): 1940-1944.
[5]	温东旭, 张雷, 索朗斯珠, 牛家强, 王玉恒, 贡嘎, 旦巴次仁, 席广银, 郭敏, 徐业芬. 牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究[J]. 畜牧兽医学报, 2019, 50(8): 1576-1586.
[6]	温东旭, 张雷, 索朗斯珠, 牛家强, 王玉恒, 贡嘎, 旦巴次仁, 席广银, 郭敏, 徐业芬. 牦牛Nramp1基因mRNA组织表达谱及其相关miRNAs初步研究[J]. 畜牧兽医学报, 2019, 50(8): 1576-1586.
[7]	施会彬, 王玉琴, 张小辉, 吴至博, 杨芳. 基于转录组测序数据的山羊下丘脑组织繁殖相关信号通路及候选基因的筛选[J]. 畜牧兽医学报, 2019, 50(7): 1380-1395.
[8]	施会彬, 王玉琴, 张小辉, 吴至博, 杨芳. 基于转录组测序数据的山羊下丘脑组织繁殖相关信号通路及候选基因的筛选[J]. 畜牧兽医学报, 2019, 50(7): 1380-1395.
[9]	许涛, 潘阳阳, 何翃闳, 李谷月, 张慧珠, 赵凌, 崔燕, 余四九. TNF-α对牦牛卵母细胞HIF-1α和HSP70的表达及后续胚胎发育能力的影响[J]. 畜牧兽医学报, 2019, 50(6): 1198-1207.
[10]	许涛, 潘阳阳, 何翃闳, 李谷月, 张慧珠, 赵凌, 崔燕, 余四九. TNF-α对牦牛卵母细胞HIF-1α和HSP70的表达及后续胚胎发育能力的影响[J]. 畜牧兽医学报, 2019, 50(6): 1198-1207.
[11]	纪会, 王会, 柴志欣, 王吉坤, 王嘉博, 罗晓林, 姬秋梅, 信金伟, 钟金城. 牦牛不同年龄肌肉组织microRNA表达谱及生物信息学分析[J]. 畜牧兽医学报, 2019, 50(5): 957-971.
[12]	张石磊, 包佳鹭, 史万玉, 钟秀会. 母鼠孕期和哺乳期连续染毒双酚A对子代雄鼠睾丸发育的影响[J]. 畜牧兽医学报, 2019, 50(5): 993-1005.
[13]	纪会, 王会, 柴志欣, 王吉坤, 王嘉博, 罗晓林, 姬秋梅, 信金伟, 钟金城. 牦牛不同年龄肌肉组织microRNA表达谱及生物信息学分析[J]. 畜牧兽医学报, 2019, 50(5): 957-971.
[14]	张石磊, 包佳鹭, 史万玉, 钟秀会. 母鼠孕期和哺乳期连续染毒双酚A对子代雄鼠睾丸发育的影响[J]. 畜牧兽医学报, 2019, 50(5): 993-1005.
[15]	刘丽娟, 董然然, 王开功, 程振涛, 文明, 温贵兰, 李晨, 杨琦, 周碧君. 敲除小RNA gcvB后沙门菌的转录组分析[J]. 畜牧兽医学报, 2019, 50(4): 840-850.