鸡lncRNA-MSTRG.15568.9及其预测靶基因的表达

doi:10.16431/j.cnki.1671-7236.2019.07.013

摘要/Abstract

摘要：

试验旨在了解在鸡睾丸中高表达的1个长链非编码RNA （long non-coding RNA，lncRNA）及其预测靶基因的时空表达规律，研究二者在鸡弱精子症中的调控作用。根据弱精子症和正常北京油鸡公鸡睾丸转录组测序筛选到的1个高表达的lncRNA （MSTRG.15568.9），采用顺式（cis）作用模式预测其潜在靶基因SPAG4（sperm-associated antigen 4），进一步采用实时荧光定量PCR方法进行表达量分析。分别选择3只0、5、20、30、45、60周龄正常北京油鸡公鸡，检测MSTRG.15568.9与SPAG4基因在不同周龄公鸡睾丸中的表达量差异；选择30周龄3只正常公鸡，采集睾丸、肝脏和脾脏等8个部位组织样品，检测MSTRG.15568.9与SPAG4基因在不同组织间的表达规律；选择45周龄弱精子症公鸡和正常公鸡各3只，对比MSTRG.15568.9与SPAG4基因在睾丸的表达量差异。结果显示，MSTRG.15568.9与SPAG4存在明显的时空表达差异，且二者表达趋势基本一致。在不同周龄的鸡睾丸组织中，MSTRG.15568.9和SPAG4的表达趋势相近，MSTRG.15568.9在20周龄的表达量显著高于0、5、30、45、60周龄（P<0.05），0和5周龄表达量显著低于20、30、45和60周龄（P<0.05）；SPAG4在45周龄表达量最高，其次是20周龄（P<0.05）。MSTRG.15568.9和SPAG4在睾丸和肝脏中的表达量均显著高于脾脏、肾脏等组织（P<0.05）；在正常睾丸组织中的表达量均显著高于弱精子症睾丸组织（P<0.05）。综上所述，MSTRG.15568.9与SPAG4基因具有较明显的组织表达特异性，且MSTRG.15568.9可能调控SPAG4基因的表达，参与精子发生与精子活力调控；但其具体作用机制需要进一步探索。本研究可为鉴定与鸡弱精子症调节机制相关的功能基因提供参考。

关键词: 鸡; lncRNA; 睾丸; 精子活力; 实时荧光定量PCR

Abstract:

The objective of this study was to study the spatio-temporal expression of a long non-coding RNA (lncRNA) and its predicted target gene,which are highly expressed in chicken testis,and its regulation in asthenospermia Beijing-You chickens.A high-expression lncRNA (MSTRG.15568.9) was screened by the transcriptome of normal and the asthenospermia Beijing-You chickens.Its potential target gene (sperm-associated antigen 4,SPAG4) was revealed by cis prediction,and RT-qPCR was used to further expression analysis.3 normal cocks with similar body weight were randomly selected at 0,5,20,30,45,and 60 weeks of age to detect the difference in expression between MSTRG.15568.9 and SPAG4 at different ages;At 30 weeks of age,3 normal cocks were selected,and 8 tissues including testis,liver and spleen etc. collected to detect the expression of MSTRG.15568.9 and SPAG4.At 45 weeks of age,3 normal and 3 asthenospermic cocks were selected,and the difference in expression levels between MSTRG.15568.9 and SPAG4 were compared.The results showed that MSTRG.15568.9 and SPAG4 had significant spatio-temporal expression differences,and the expression trends of them were basically the same.The expression trends of MSTRG.15568.9 and SPAG4 were similar in testis tissues of different ages.The expression level of MSTRG.15568.9 at 20 weeks was significantly higher than that at 0,5,30,45 and 60 weeks (P<0.05).The expression levels at 0 and 5 weeks of age were significantly lower than those at 20,30,45 and 60 weeks (P<0.05);SPAG4 was highest at 45 weeks of age,followed by 20 weeks (P<0.05).The expression levels of MSTRG.15568.9 and SPAG4 in testis and liver were significantly higher than that in spleen and kidney (P<0.05).The expression levels of MSTRG.15568.9 and SPAG4 in testis of cocks with normal sperm motility were significantly higher than those in asthenospermia cocks (P<0.05).In conclusion,MSTRG.15568.9 and SPAG4 had obvious tissue expression specificity,and the expression of SPAG4 might be related to MSTRG.15568.9,and both could be involved in spermatogenesis and sperm motility regulation.The specific mechanisms need to be further explored.This study could provide references for the identification of functional genes related to the regulation mechanism of chicken asthenospermia.

Key words: chicken; lncRNA; testis; sperm motility; RT-qPCR

中图分类号:

S831.2

黄子妍, 孙研研, 李云雷, 许红, 刘一帆, 石雷, 叶建华, 麻慧, 倪爱心, 王攀林, 谢金防, 陈继兰. 鸡lncRNA-MSTRG.15568.9及其预测靶基因的表达[J]. 《中国畜牧兽医》, 2019, 46(7): 1986-1993.

HUANG Ziyan, SUN Yanyan, LI Yunlei, XU Hong, LIU Yifan, SHI Lei, YE Jianhua, MA Hui, NI Aixin, WANG Panlin, Xie Jinfang, CHEN Jilan. Expression of lncRNA-MSTRG.15568.9 and Its Predicted Target Gene in Chicken[J]. , 2019, 46(7): 1986-1993.

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