三个阶段牦牛睾丸发育过程中piRNA的鉴定及分析研究

doi:10.11843/j.issn.0366-6964.2019.11.007

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (11): 2235-2243.doi: 10.11843/j.issn.0366-6964.2019.11.007

三个阶段牦牛睾丸发育过程中piRNA的鉴定及分析研究

殷实^1,2,3, 秦文昌^1,2,3, 王斌^1,2,3, 周婧雯^1,2,3, 杨柳青^1,2,3, 李键^1,2,3*

1. 西南民族大学生命科学与技术学院, 成都 610041;
2. 青藏高原动物遗传资源保护与利用教育部重点实验室, 成都 610041;
3. 青藏高原动物遗传资源保护与利用四川省重点实验室, 成都 610041

收稿日期:2019-05-22 出版日期:2019-11-23 发布日期:2019-11-23
通讯作者: 李键,主要从事牦牛细胞生物学和发育生物学研究,E-mail:jianli_1967@163.com
作者简介:殷实(1988-),男,河南焦作人,博士,讲师,主要从事动物细胞与胚胎工程研究,E-mail:raulyinshi@163.com
基金资助:
中央高校基本科研业务费专项基金项目（2018NQN33）；四川省科技支撑计划（2017NZ0076）；牦牛遗传资源与保护创新团队（13CXTD01）；西南民族大学研究生创新型科研项目（CX2018SZ10）；青藏高原生态畜牧业协同创新中心开放基金（QZGYXT05）；青藏高原动物遗传资源保护与利用重点实验室

Identification and Analysis of Relevant piRNA during Testicular Development in Yak of Three Stages

YIN Shi^1,2,3, QIN Wenchang^1,2,3, WANG Bin^1,2,3, ZHOU Jingwen^1,2,3, YANG Liuqing^1,2,3, LI Jian^1,2,3*

1. College of Life Science and Technology, Southwest Minzu University, Chengdu 610041, China;
2. Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization of Ministry of Education, Chengdu 610041, China;
3. Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization Key Laboratory of Sichuan Province, Chengdu 610041, China

Received:2019-05-22 Online:2019-11-23 Published:2019-11-23

摘要/Abstract

摘要： 旨在对不同年龄牦牛睾丸发育过程中的piRNA进行鉴定及分析。本研究选择胎牛期（4～5个月）、幼年期（1岁）及青年期（3岁）健康雄性牦牛共6头（每组各两头），分离其睾丸进行piRNA测序，其中来源于同一年龄的两个组织样本视为生物学重复。分析样本中piRNA的数量、碱基偏好性、来源、功能、piRNA簇的染色体分布及表达水平，并利用荧光定量PCR检测3个阶段牦牛睾丸中PIWI家族（PIWIL1～PIWIL4）的表达。结果表明，幼年期及青年期牦牛睾丸中piRNA的数量均显著多于胎牛阶段piRNA的数量（P<0.01）。牦牛睾丸piRNA 5'端具有明显的尿嘧啶（U）偏好性。大部分的piRNA来自基因间区和基因区之外的其他区域。胎牛睾丸中超过60%的piRNA簇处于低丰度，而幼年期及青年期牦牛睾丸组织中超过70%的piRNA簇均处于高丰度。胎牛期睾丸中PIWIL1及PIWIL4的表达与其他两个时期相比存在显著差异（P<0.05）。GO分析发现，piRNA来源基因的数目在生物学过程、细胞组分和分子功能上排在首位的分别为代谢过程、细胞和结合。结果提示，牦牛睾丸中piRNA的结构、功能和来源具有特异性。胚后阶段（幼年期及青年期）牦牛睾丸piRNA的数量及表达丰度与胚胎期相比有显著差异，这可能是与PIWIL1及PIWIL4在两个阶段表达的差异有关。本研究为进一步探索piRNA调控牦牛睾丸的发育机制，以及改善牦牛的生产性能提供了一定的理论基础。

Abstract: This research was conducted to identify and analyze piRNA in the testicular development of yaks of different ages. A total of 6 healthy male yaks (two in each group) in the fetal (4-5 months old), calves (1 year old) and juvenile (3 years old) stages were selected. Testis were separated for piRNA sequencing and two samples from yak at the same age were considered as biological duplications. The number, base preference, source and function of piRNA, as well as the chromosome distribution and expression levels of piRNA clusters in the samples were analyzed. The expression of PIWI gene family(PIWIL1-PIWIL4) in the testis of yak at 3 stages were detected by fluorescence quantitative PCR. The results showed that the number of piRNAs in testis of yaks in calves and juvenile was significantly more than that in fetal stage (P<0.01). piRNAs from yak testis showed a strong preference for uracil (U) at their 5' ends, and the most of the piRNA came from the other regions than the intergenic and gene regions. More than 60% piRNA clusters in fetal testes were at low abundance, while more than 70% clusters in testes of calves and juvenile yaks were at high abundance. The expressions of PIWIL1 and PIWIL4 in fetal testes were significantly different compared with those in the other two stages (P<0.05). GO function analysis result showed that the number of piRNA source genes ranked first in biological process, cell component and molecular function were metabolic process, cell and binding, respectively. The structure, function and source of piRNA in yak testes were specific. The number and expression abundance of piRNA in the post-embryonic stage (calves and juvenile) was significantly different from that in the fetal stage, which may be related to the expression difference of PIWIL1 and PIWIL4 between two stages.This research provides a theoretical basis for further studying the mechanism of piRNA regulating yak testicular development and improving yak production performance.

中图分类号:

S823.85

殷实, 秦文昌, 王斌, 周婧雯, 杨柳青, 李键. 三个阶段牦牛睾丸发育过程中piRNA的鉴定及分析研究[J]. 畜牧兽医学报, 2019, 50(11): 2235-2243.

YIN Shi, QIN Wenchang, WANG Bin, ZHOU Jingwen, YANG Liuqing, LI Jian. Identification and Analysis of Relevant piRNA during Testicular Development in Yak of Three Stages[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(11): 2235-2243.

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参考文献 30

[1]	韩杰,熊显荣,蔡雯祎,等.牦牛KDM4A基因克隆、组织表达谱及其在卵母细胞和颗粒细胞中的表达[J].畜牧兽医学报, 2018,49(2):291-299.
	HAN J,XIONG X R,CAI W Y,et al.Cloning of KDM4A gene and its expression in different tissues,oocyte and granulosa cell of yak[J].Acta Veterinaria et Zootechnica Sinica,2018, 49(2):291-299.(in Chinese)
[2]	蔡雯祎,熊显荣,陈通,等.KDM2B基因克隆及其在牦牛组织和卵母细胞减数分裂过程中的表达研究[J].畜牧兽医学报, 2018,49(3):534-541.
	CAI W Y,XIONG X R,CHEN T,et al.Molecular cloning of KDM2B gene and its expression pattern in tissues and the process of oocyte meiosis in yak[J].Acta Veterinaria et Zootechnica Sinica,2018,49(3):534-541.(in Chinese)
[3]	LI B,HE X L,ZHAO Y P,et al.Identification of piRNAs and piRNA clusters in the testes of the Mongolian horse[J].Sci Rep,2019, 9:5022.
[4]	CHALBATANI G M,DANA H,MEMARI F,et al.Biological function and molecular mechanism of piRNA in cancer[J].Pract Lab Med,2019,13:e00113.
[5]	STURM Á,PERCZEL A,IVICS Z,et al.The Piwi-piRNA pathway:road to immortality[J].Aging Cell,2017,16(5):906-911.
[6]	HAN B W,ZAMORE P D.piRNAs[J].Curr Biol,2014,24(16):R730-R733.
[7]	ROSENKRANZ D.piRNA cluster database:a web resource for piRNA producing loci[J].Nucleic Acids Res,2016,44(D1):D223-D230.
[8]	HUANG Y,BAI J Y,REN H T.piRNAs biogenesis and its functions[J].Bioorg Khim,2014,40(3):320-326.
[9]	CZECH B,HANNON G J.One loop to rule them all:the ping-pong cycle and piRNA-guided silencing[J].Trends Biochem Sci, 2016,41(4):324-337.
[10]	HAN B W,WANG W,LI C J,et al.piRNA-guided transposon cleavage initiates Zucchini-dependent,phased piRNA production[J]. Science,2015,348(6236):817-821.
[11]	HAYASHI R,SCHNABL J,HANDLER D,et al.Genetic and mechanistic diversity of piRNA 3'-end formation[J].Nature, 2016,539(7630):588-592.
[12]	GRIVNA S T,BEYRET E,WANG Z,et al.A novel class of small RNAs in mouse spermatogenic cells[J].Genes Dev,2006, 20(13):1709-1714.
[13]	WATANABE T,TAKEDA A,TSUKIYAMA T,et al.Identification and characterization of two novel classes of small RNAs in the mouse germline:retrotransposon-derived siRNAs in oocytes and germline small RNAs in testes[J].Genes Dev,2006,20(13):1732-1743.
[14]	MOYANO M,STEFANI G.piRNA involvement in genome stability and human cancer[J].J Hematol Oncol,2015,8:38.
[15]	GIRARDI E,MIESEN P,PENNINGS B,et al.Histone-derived piRNA biogenesis depends on the ping-pong partners Piwi5 and Ago3 in Aedes aegypti[J].Nucleic Acids Res,2017,45(8):4881-4892.
[16]	NEWKIRK S J,LEE S,GRANDI F C,et al.Intact piRNA pathway prevents L1 mobilization in male meiosis[J].Proc Natl Acad Sci U S A,2017,114(28):E5635-E5644.
[17]	WU S R,GUO W,YAN T,et al.Spermatozoal mRNAs expression implicated in embryonic development were influenced by dietary folate supplementation of breeder roosters by altering spermatozoal piRNA expression profiles[J].Theriogenology, 2019,138:102-110.
[18]	DAI X Y,SHU Y Q,LOU Q Y,et al.Tdrd12 is essential for germ cell development and maintenance in Zebrafish[J].Int J Mol Sci,2017,18(6):1127.
[19]	MANAKOV S A,PEZIC D,MARINOV G K,et al.MIWI2 and MILI have differential effects on piRNA biogenesis and DNA methylation[J].Cell Rep,2015,12(8):1234-1243.
[20]	THÉRON E,MAUPETIT-MEHOUAS S,POUCHIN P,et al.The interplay between the Argonaute proteins Piwi and Aub within Drosophila germarium is critical for oogenesis,piRNA biogenesis and TE silencing[J].Nucleic Acids Res,2018,46(19):10052-10065.
[21]	CANNARELLA R,CONDORELLI R A,DUCA Y,et al.New insights into the genetics of spermatogenic failure:a review of the literature[J].Hum Genet,2019,138(2):125-140.
[22]	SUTHERLAND J M,SOBINOFF A P,FRASER B A,et al.RNA binding protein Musashi-2 regulates PIWIL1 and TBX1 in mouse spermatogenesis[J].J Cell Physiol,2018,233(4):3262-3273.
[23]	LI Y,LI J L,FANG C C,et al.Genome-wide differential expression of genes and small RNAs in testis of two different porcine breeds and at two different ages[J].Sci Rep,2016,6:26852.
[24]	ZHANG Y,WANG X H,KANG L.A k-mer scheme to predict piRNAs and characterize locust piRNAs[J].Bioinformatics, 2011,27(6):771-776.
[25]	徐璐.鸡精子发生过程中关键piRNAs及Piwill基因功能初步研究[D].扬州:扬州大学,2017.
	XU L.Prelimianary study on the biological function of piRNAs and Piwil1gene during spermatogenesis in poultry[D].Yangzhou:Yangzhou University,2017.(in Chinese)
[26]	王标,付绍印,何小龙,等.绵羊睾丸及卵巢组织中piRNA表达情况的测序分析[C]//2017年全国养羊生产与学术研讨会暨养羊学分会第七次全国会员代表大会.石家庄:中国畜牧兽医学会养羊学分会,2017.
	WANG B,FU S Y,HE X L,et al.Sequencing analysis of the expression of piRNAs in the testes and ovaries of sheep[C]//2017 National Seminar on Sheep Production and Academic Research and the Seventh National Congress of the Society of Sheep Science.Shijia-zhuang:Society of Sheep Breeding, Chinese Society of Animal Husbandry and Veterinary Medicine,2017.(in Chinese)
[27]	FUJIHARA Y,OJI A,LARASATI T,et al.Human globozoospermia-related gene Spata16 is required for sperm formation revealed by CRISPR/Cas9-mediated mouse models[J].Int J Mol Sci,2017,18(10):2208.
[28]	SINGH A P,RAJENDER S.CatSper channel,sperm function and male fertility[J].Reprod BioMed Online,2015,30(1):28-38.
[29]	YAMTICH J,HEO S J,DHAHBI J,et al.piRNA-like small RNAs mark extended 3'UTRs present in germ and somatic cells[J].BMC Genomics,2015,16:462.
[30]	STAUB C,JOHNSON L.Review:Spermatogenesis in the bull[J].Animal,2018,12(S1):S27-S35.

三个阶段牦牛睾丸发育过程中piRNA的鉴定及分析研究

Identification and Analysis of Relevant piRNA during Testicular Development in Yak of Three Stages

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