分子与细胞营养 MOLECULAR AND CELLULAR NUTRITION

硒代蛋氨酸对种公鸡睾丸基因差异表达的影响及功能预测分析

  • 衡诺 ,
  • 马鑫 ,
  • 郭勇 ,
  • 齐晓龙 ,
  • 盛熙晖 ,
  • 王相国 ,
  • 邢凯 ,
  • 肖龙菲 ,
  • 陈余 ,
  • 王梁 ,
  • 倪和民
展开
  • 1. 北京农学院动物科学技术学院, 北京 102206;
    2. 北京市畜牧总站, 北京 100107
衡诺(1995-),男,北京人,硕士研究生,从事畜禽营养与动物繁殖育种研究。E-mail:1017337574@qq.com

收稿日期: 2021-01-24

  网络出版日期: 2021-08-11

基金资助

现代农业产业技术体系北京市家禽创新团队项目(BAJC04-2021);国家"十三五"重点研发计划(2016YFD0700201)

Effects of Selenomethionine on Gene Differential Expression in Testis of Breeding Cocks and Function Prediction

  • HENG Nuo ,
  • MA Xin ,
  • GUO Yong ,
  • QI Xiaolong ,
  • SHENG Xihui ,
  • WANG Xiangguo ,
  • XING Kai ,
  • XIAO Longfei ,
  • CHEN Yu ,
  • WANG Liang ,
  • NI Hemin
Expand
  • 1. College of Animal Science and Technology, Beijing Agricultural University, Beijing 102206, China;
    2. Beijing General Station of Animal Husbandry, Beijing 100107, China

Received date: 2021-01-24

  Online published: 2021-08-11

Supported by

 

摘要

本试验以配种后期种公鸡为动物模型,比较对照组和有机硒组睾丸组织基因的差异表达,结合GO和KEGG功能富集性分析,旨在探讨硒对种公鸡繁殖能力的影响,为后续提高种公鸡繁殖性能的营养调控提供理论支撑。试验选取120只378日龄健康、体况一致的罗曼褐蛋用种公鸡,随机分为2个组,每组6个重复,每个重复10只鸡。对照组饲喂基础饲粮,试验组在基础饲粮中添加硒水平为1 mg/kg的硒代蛋氨酸。试验预试期1周,正试期4周。试验期末,从每个重复中随机挑选1只种公鸡取其左侧睾丸,进行睾丸转录组测序,筛选睾丸组织中显著差异表达的基因。结果表明,通过对2组种公鸡的睾丸组织样本进行转录组测序,总共挖掘出111个显著差异表达基因,包括55个上调基因和56个下调基因,注释到的基因个数为86个。在这些差异表达的基因中,上调差异表达倍数最大的基因是微管蛋白alpha-8链(TUBA8A),下调差异表达倍数最大的基因是钙/钙调蛋白依赖性蛋白激酶Ⅳ样(CAMK4L)。相对荧光定量PCR结果与睾丸组织转录组测序结果表达趋势相似。GO功能分析显示,蛋白质chibby同源物1(CBY1)、TUBA8AADP核糖基转移酶4(ART4)、钙周期蛋白结合蛋白(CACYBP)、腺苷酸激酶7(AK7)和线粒体核糖体蛋白L17(MRPL17)等基因通过参与细胞过程、生物调节、代谢过程、细胞组分和分子功能等提高种公鸡繁殖性能,可作为研究种公鸡繁殖性能的候选基因并进行进一步地研究和探讨。

本文引用格式

衡诺 , 马鑫 , 郭勇 , 齐晓龙 , 盛熙晖 , 王相国 , 邢凯 , 肖龙菲 , 陈余 , 王梁 , 倪和民 . 硒代蛋氨酸对种公鸡睾丸基因差异表达的影响及功能预测分析[J]. 动物营养学报, 2021 , 33(8) : 4626 -4636 . DOI: 10.3969/j.issn.1006-267x.2021.08.042

Abstract

In this experiment, breeding cocks in the late breeding period were used as animal models to compare the gene differential expression in testis tissues between the control group and the organic selenium group. Combined with GO and KEGG functional enrichment analysis, the aim of this study was to explore the effects of selenium on reproductive performance of breeding cocks, and to provide theoretical support for nutritional regulation to improve reproductive performance of breeding cocks in the future. A total of 120 healthy Loman brown breeding cocks with the same body condition at the age of 378 days were randomly divided into 2 groups with 6 replicates per group and 10 cocks per replicate. Cocks in the control group were fed a basal diet, and those in the experimental group were fed the basal diet supplemented with selenomethionine at a selenium level of 1 mg/kg. The pre-trial period lasted for 1 week and the experimental period lasted for 4 weeks. At the end of the experiment period, one of the cocks from each replicate was randomly selected to take the left testis. The testis transcriptome was sequenced to screen significantly expressed genes in the testis tissues. The results showed that a total of 111 differentially expressed genes, including 55 up-regulated genes and 56 down-regulated genes, were found by transcriptome sequencing in testis tissue samples of cocks from two groups, and the number of annotated genes was 86. Among the differentially expressed genes, tubulin alpha-8 chain (TUBA8A) gene had the highest up-regulated differential expression, while calcium/calmodulin-dependent protein kinase Ⅳ like (CAMK4L) had the highest down-regulated differential expression. The expression trend of RT-PCR was similar to that of transcriptome sequencing in testis tissues. GO function analysis showed that protein chibby homology 1 (CBY1), TUBA8A, ADP-ribosyltransferase 4 (ART4), calcyclin binding protein (CACYBP), adenylate kinase 7 (AK7) and mitochondrial ribosomal protein L17 (MRPL17) genes were involved in cellular processes, biological regulation, metabolic processes, cellular components and molecular function to improve the reproductive performance of breeding cocks, and there can be used as candidate genes to study the reproductive performance of cocks for further study and discussion.

参考文献

[1] AVERY J C,HOFFMANN P R.Selenium,selenoproteins,and immunity[J].Nutrients,2018,10(9):1203.
[2] 梁明振,峗新跃,黄钦华,等.微量元素硒对动物繁殖性能的影响[J].中国畜牧兽医,2003,30(3):14-16. LIANG M Z,WEI X Y,HUANG X H,et al.The effect of microelement selenium on animal reproductive performance[J].China Animal Husbandry & Veterinary Medicine,2003,30(3):14-16.(in Chinese)
[3] MEHDI Y,HORNICK J L,ISTASSE L,et al.Selenium in the environment,metabolism and involvement in body functions[J].Molecules,2013,18(3):3292-3311.  
[4] RAYMAN M P.Selenium and human health[J].The Lancet,2012,379(9822):1256-1268.  
[5] WALDNER C L,VAN DE WEYER L M.Selenium status at the end of the grazing season,reproductive performance and degenerative myopathy in beef herds[J].The Canadian Veterinary Journal,2011,52(10):1083-1088.
[6] KHALIL-KHALILI A A,ZHANDI M,ZAGHARI M,et al.The effect of dietary organic selenium on reproductive performance of broiler breeder roosters under dexamethasone induced stress[J].Theriogenology,2020,161:16-25.
[7] 朱冠宇,李征,张立昌,等.硒代蛋氨酸对蛋用种公鸡繁殖性能及血液生殖激素的影响[J].黑龙江畜牧兽医,2017(11):1-4. ZHU G Y,LI Z,ZHANG L C,et al.Effects of selenomethionine on reproductive performance and blood reproductive hormone in breeder cocks[J].Heilongjiang Animal Science and Veterinary Medicine,2017(11):1-4.(in Chinese)
[8] KIM D,PERTEA G,TRAPNELL C,et al.TopHat2:accurate alignment of transcriptomes in the presence of insertions,deletions and gene fusions[J].Genome Biology,2013,14(4):R36.
[9] 马鑫,李润华,盛熙辉,等.硒代蛋氨酸对种公鸡精液品质及精浆抗氧化能力的影响[J].安徽农业科学,2019,47(17):94-97. MA X,LI R H,SHENG X H,et al.Effects of selenomethionine on the semen quality and antioxidant capacity of seminal plasma of breeding cocks[J].Journal of Anhui Agricultural Sciences,2019,47(17):94-97.(in Chinese)
[10] LEE Y L,SANTE J,COMERCI C J,et al.Cby1 promotes Ahi1 recruitment to a ring-shaped domain at the centriole-cilium interface and facilitates proper cilium formation and function[J].Molecular Biology of the Cell,2014,25(19):2919-2933.  
[11] FISCHER V,WONG M,LI F Q,et al.Chibby1 knockdown promotes mesenchymal-to-epithelial transition-like changes[J].Cell Cycle,2017,16(5):448-456.  
[12] GRAHNERT A,RICHTER S,SIEGERT F,et al.The orthologue of the "acatalytic" mammalian ART4 in chicken is an arginine-specific mono-ADP-ribosyltransferase[J].BMC Molecular Biology,2008,9:86.
[13] 杨春燕,张文,付薇,等.ADP核糖基化对DNA损伤修复的调控[J].现代农业科技,2015(18):273-274,282. YANG C Y,ZHANG W,FU W,et al.ADP-ribosylation signaling during DNA damage repair[J].Modern Agricultural Science and Technology,2015(18):273-274,282.(in Chinese)
[14] CHAPEL-FERNANDES S,MOVIA C,JORDIER F,et al.DO/ART4 gene sequencing in sub-Saharan cohorts and African migrants:useful data describing the diversity and spreading of rare variants[J].Transfusion,2019,59(12):3755-3766.  
[15] MORRIS S J Jr.Arginine Metabolism Revisited[J].The Journal of Nutrition,2016,146(12):2579S-2586.
[16] MACIEL JR V L,CALDAS-BUSSIERE M C,SILVEIRA V,et al.L-arginine alters the proteome of frozen-thawed bovine sperm during in vitro capacitation[J].Theriogenology,2018,119:1-9.
[17] IVLIEV A E,'T HOEN PETER A C,VAN ROON-MOM W M C,et al.Exploring the transcriptome of ciliated cells using in silico dissection of human tissues[J].PLoS One,2012,7(4):e35618.
[18] FERNANDEZ-GONZALEZ A,KOUREMBANAS S,WYATT T A,et al.Mutation of murine adenylate kinase 7 underlies a primary ciliary dyskinesia phenotype[J].American Journal of Respiratory Cell and Molecular Biology,2009,40(3):305-313.  
[19] MILARA J,ARMENGOT M,MATA M,et al.Role of adenylate kinase type 7 expression on cilia motility:possible link in primary ciliary dyskinesia[J].American Journal Of Rhinology & Allergy,2010,24(3):181-185.  
[20] MATA M,LLUCH-ESTELLÉS J,ARMENGOT M,et al.New adenylate kinase 7(AK7) mutation in primary ciliary dyskinesia[J].American Journal of Rhinology & Allergy,2012,26(4):260-264.  
[21] LORōS P,COUTTON C,EL KHOURI E,et al.Homozygous missense mutation L673P in adenylate kinase 7(AK7) leads to primary male infertility and multiple morphological anomalies of the flagella but not to primary ciliary dyskinesia[J].Human Molecular Genetics,2018,27(7):1196-1211.  
[22] BENARROCH E E.Dynamics of microtubules and their associated proteins:recent insights and clinical implications[J].Neurology,2016,86(20):1911-1920.  
[23] 李建农,蒋建东.微管的生物学特性与药物研究[J].药学学报,2003,38(4):311-315. LI J N,JIANG J D.Biological characteristics of microtubule and related drug research[J].Acta Pharmaceutica Sinica,2003,38(4):311-315.(in Chinese)
[24] GOODSON H V,JONASSON E M.Microtubules and microtubule-associated proteins[J].Cold Spring Harbor Perspectives in Biology,2018,10(6):a022608.doi:10.1101/cshperspect.a022608.
[25] BOX J M,KAUR J,STUART R A.MrpL35,a mitospecific component of mitoribosomes,plays a key role in cytochrome C oxidase assembly[J].Molecular Biology of the Cell,2017,28(24):3489-3499.  
[26] WERNER A,MEINHARDT A,SEITZ J,et al.Distribution of heat-shock protein 60 immunoreactivity in testes of infertile men[J].Cell and Tissue Research,1997,288(3):539-544.  
[27] DORJI,OHKUBO Y,MIYOSHI K,et al.Gene expression differences in oocytes derived from adult and prepubertal Japanese black cattle during in vitro maturation[J].Reproduction in Domestic Animals,2012,47(3):392-402.  
[28] NASRALLAH C M,HORVATH T L.Mitochondrial dynamics in the central regulation of metabolism[J].Nature Reviews.Endocrinology,2014,10(11):650-658.  
[29] TVRK G,ÇERIBASI A O,SAKIN F,et al.Antiperoxidative and anti-apoptotic effects of lycopene and ellagic acid on cyclophosphamide-induced testicular lipid peroxidation and apoptosis[J].Reproduction,Fertility And Development,2010,22(4):587-596.  
[30] MATSUZAWA S I,LI C L,NI C Z,et al.Structural analysis of Siah1 and its interactions with Siah-interacting protein (SIP)[J].Journal Biological Chemical,2003,278(3):1837-1840.  
[31] LIU J,STEVENS J,ROTE C A,et al.Siah-1 mediates a novel β-catenin degradation pathway linking p53 to the adenomatous polyposis coli protein[J].Molecular Cell,2001,7(5):927-936.  
[32] YANG Y J,LIU W M,ZHOU J X,et al.Expression and hormonal regulation of calcyclin-binding protein (CacyBP) in the mouse uterus during early pregnancy[J].Life Sciences,2006,78(7):753-760.  
[33] MATSUZAWA S I,REED J C.Siah-1,SIP,and Ebi collaborate in a novel pathway for β-catenin degradation linked to p53 responses[J].Molecular Cell,2001,7(5):915-926.  
[34] SCHNEIDER G,NIEZNANSKI K,KILANCZYK E,et al.CacyBP/SIP interacts with tubulin in neuroblastoma NB2a cells and induces formation of globular tubulin assemblies[J].Biochimica et Biophysica Acta:Molecular Cell Research,2007,1773(11):1628-1636.  
文章导航

/