去势和非去势公猪背最长肌circRNA差异表达分析

doi:10.16288/j.yczz.21-162

遗传 ›› 2021, Vol. 43 ›› Issue (11): 1066-1077.doi: 10.16288/j.yczz.21-162

去势和非去势公猪背最长肌circRNA差异表达分析

邢宝松¹(), 王璟¹(), 陈俊峰¹, 马强¹, 任巧玲¹, 张家庆¹, 张华¹, 滑留帅¹, 孙加节², 曹海³

1. 河南省农业科学院畜牧兽医研究所,河南省畜禽繁育与营养调控重点实验室,郑州 450002
2. 华南农业大学动物科学学院,广东省动物营养调控重点实验室/国家生猪种业工程技术中心,广州 510642
3. 河南兴锐农牧科技有限公司,信阳 465550

收稿日期:2021-04-27 修回日期:2021-07-28 出版日期:2021-11-20 发布日期:2021-08-24
通讯作者: 王璟 E-mail:bsxing@126.com;wangjing_0407@163.com
作者简介:邢宝松,博士,副研究员,研究方向：猪的育种与管理。E-mail: bsxing@126.com
基金资助:
国家自然科学基金青年基金项目,河南省农业科学院科技创新创意项目和河南省重点研发与推广专项资助(31601927);河南省农业科学院科技创新创意项目编号(2020CX18);河南省重点研发与推广专项编号资助(212102110010)

Analysis of differentially expressed circRNAs in longissimus muscle between castrated and intact male pigs

Baosong Xing¹(), Jing Wang¹(), Junfeng Chen¹, Qiang Ma¹, Qiaoling Ren¹, Jiaqing Zhang¹, Hua Zhang¹, Liushuai Hua¹, Jiajie Sun², Hai Cao³

1. Henan Key Laboratory of Farm Animal Breeding and Nutritional Regulation, Institute of Animal Husbandry and Veterinary Science, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China
2. Guangdong Provincial Key Laboratory Of Animal Nutrition Control/National Engineering Research Center For Breeding Swine Industry, College Of Animal Science, South China Agricultural University, Guangzhou 510642, China
3. Henan Xing Rui agricultural and animal husbandry technology Co., LTD, Xinyang 465550, China

Received:2021-04-27 Revised:2021-07-28 Online:2021-11-20 Published:2021-08-24
Contact: Wang Jing E-mail:bsxing@126.com;wangjing_0407@163.com
Supported by:
Supported by the National Natural Science Foundation of China(31601927);Technological Innovation and Creative Project from the Henan Academy of Agricultural Sciences No(2020CX18);Financial Budget Project of Henan Province No(212102110010)

摘要/Abstract

摘要：

公猪去势可减少异味和打斗,但去势后产肉量和肌内脂肪沉积发生变化,其分子机制的解析对生产具有重要意义。近年来研究表明,环状RNA (circRNA)在肌肉发育中具有重要调控作用。为探究去势后circRNAs对背最长肌发育的调控,本研究选择6头淮南公猪,随机选择3头去势,当体重达130 kg左右屠宰,采集背最长肌样品,利用高通量测序筛选差异表达circRNAs (differentially expressed circRNAs, DECs)并进行KEGG功能富集分析。结合前期筛选的公猪去势相关miRNAs,构建DECs-miRNAs调控网络,最后使用猪骨骼肌卫星细胞验证候选circRNA表达谱及其与miRNA互作关系。结果表明,去势和非去势组背最长肌样品共获得5866个circRNAs,两组之间共有370个DECs (| log₂Foldchange | > 1, p_adj<0.8),KEGG富集分析表明,DECs来源母基因主要富集于肌肉发育、肌纤维类型转化、能量代谢等相关通路。构建的DECs-miRNA调控网络共包括69个circRNAs和8个miRNAs。选择circRNA_2241和circRNA_4237进行验证,结果发现这两个circRNAs真实存在且表达趋势与测序结果一致。进一步在猪骨骼肌卫星细胞初步验证circRNA_2241与miR-1互作关系,结果表明睾酮显著促进circRNA_2241表达,同时抑制miR-1表达。本研究结果提示circRNAs可能通过与miRNAs互作调控猪去势后背最长肌发育,从而为解析去势对肌肉发育调控的分子机制提供参考。

关键词: 环状RNA, 去势, 公猪, 背最长肌

Abstract:

Castration can reduce odor and fights in boars, but the carcass yield is reduced, and the intramuscular fat content is increased. Understanding its molecular mechanism is of great significance for production. Recent studies have shown that circular RNAs (circRNAs) play an important role(s) in the regulation of muscle development. To explore the effects of circRNAs on the development of longissimus dorsi (LD) muscle after castration, six Huainan male pigs were selected and three of which were randomly castrated. Six pigs were slaughtered when their body weight reached around 130 kg, and the LD muscle samples were collected. The differentially expressed circRNAs (DECs) were screened by high-throughput sequencing and functionally analyzed using the KEGG databases. DECs-miRNAs network was constructed, and the expression profiles of candidate circRNAs and their interactions with miRNAs were verified in porcine skeletal muscle satellite cells. The results showed that a total of 5866 circRNAs were obtained, and 370 DECs were identified in LD muscle between the castrated and intact groups (| log₂Foldchange | > 1, p_adj <0.8). KEGG enrichment indicated that the parental genes for the DECs were mainly enriched in the pathways associated with muscle development, muscle fiber type transformation, and energy metabolism. There were 8 miRNAs and 69 circRNAs enriched in the DECs-miRNA network. circRNA_2241 and circRNA_4237 were selected for verification, which showed that these two circRNAs really existed and their expression profiles were consistent with the sequencing results. Further, preliminary analysis showed that circRNA_2241 interacted with miR-1, and testosterone promoted circRNA_2241 but inhibited miR-1 expression. These results confirmed that circRNAs might participate in the regulation of LD muscle development after castration by interacting with miRNAs, thereby providing new materials and references for analyses on the molecular mechanisms of castration on the regulation of muscle development.

Key words: circRNAs, castration, male pigs, longissimus muscle

邢宝松, 王璟, 陈俊峰, 马强, 任巧玲, 张家庆, 张华, 滑留帅, 孙加节, 曹海. 去势和非去势公猪背最长肌circRNA差异表达分析[J]. 遗传, 2021, 43(11): 1066-1077.

Baosong Xing, Jing Wang, Junfeng Chen, Qiang Ma, Qiaoling Ren, Jiaqing Zhang, Hua Zhang, Liushuai Hua, Jiajie Sun, Hai Cao. Analysis of differentially expressed circRNAs in longissimus muscle between castrated and intact male pigs[J]. Hereditas(Beijing), 2021, 43(11): 1066-1077.

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位点	引物序列(5ʹ-3ʹ)		扩增长度(bp)
位点	上游引物	下游引物	扩增长度(bp)
circRNA_2241	CAATCCTTCAGCCCAGAGAG	TGTCCAGAATGAACATCATAG	134
circRNA_4237	CTCCTAAAACCACAGGTGTG	TTGTGGATAAGGTGGGTACG	101
GAPDH	ACCAGGTTGTGTCCTGTGAC	AGCTTGACGAAGTGGTCGTT	94

类型	GO通路	GO编号	富集基因数
细胞组分	细胞内部分	GO:0044424	1732
	细胞内	GO:0005622	1813
	细胞内膜结合细胞器	GO:0043231	1369
	细胞内细胞器	GO:0043229	1492
	膜结合细胞器	GO:0043227	1498
	细胞器	GO:0043226	1605
	核部分	GO:0044428	615
	细胞内细胞器部分	GO:0044446	983
	细胞器部分	GO:0044422	993
	核	GO:0005634	882
生物过程	细胞大分子代谢过程	GO:0044260	1097
	细胞代谢过程	GO:0044237	1374
	大分子代谢过程	GO:0043170	1184
	细胞器组织	GO:0006996	503
	初级代谢过程	GO:0044238	1380
	有机物代谢过程	GO:0071704	1414
	细胞蛋白质代谢过程	GO:0044267	660
	代谢过程	GO:0008152	1600
	细胞成分的组织或生物发生	GO:0071840	769
	细胞成分组织	GO:0016043	744
分子功能	蛋白质结合	GO:0005515	1709
	杂环化合物结合	GO:1901363	801
	有机环状化合物结合	GO:0097159	804
	核酸结合	GO:0003676	466
	酶结合	GO:0019899	263
	结合	GO:0003723	143
	核苷酸结合	GO:0000166	442
	磷酸核苷结合	GO:1901265	442
	小分子结合	GO:0036094	461
	催化活性	GO:0003824	949

去势和非去势公猪背最长肌circRNA差异表达分析

Analysis of differentially expressed circRNAs in longissimus muscle between castrated and intact male pigs

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