低温胁迫下民猪骨骼肌的转录调控分析

doi:10.11843/j.issn.0366-6964.2022.08.011

摘要/Abstract

摘要： 为了揭示民猪优良种质特性的遗传机理，解析其低温适应机制。本试验将6头6月龄体重相近的雌性民猪随机分成2组，每组3头个体。对照组个体置于温度控制在（18±2）℃的猪舍内，试验组个体置于室外的半敞篷舍内饲养，环境温度从第1天的5℃/-5℃降到最后1天的-15℃/-24℃，共处理了58 d。两组个体均保证自由采食和饮水，试验结束后，屠宰全部个体，取背最长肌进行RNA-seq。筛选民猪骨骼肌受低温诱导的基因和lncRNA，并对它们进行功能注释以及两者间调控关系的分析。RNA测序分析显示，民猪在经历58 d的低温胁迫后，骨骼肌内86个基因发生了显著上调，16个基因发生了显著下调；112个lncRNAs发生了显著上调，74个lncRNAs发生了显著下调。在发生显著上调的基因中，有4个与神经系统相关的基因NTSR2、ARC、FOSL1和RCAN1发生了显著上调，7个与基质转运相关的基因SLC2A4、SLC2A5、SLC4A10、SLC19A2、SLC20A1、SLC28A1和SLC38A2，6个与炎症和免疫相关的基因AREG、CISH、OTUD1、TRIB1、GPA33和ITPKC均发生了显著上调。而与昼夜节律相关的ARNTL基因和抑制细胞增殖的RASL11A基因等发生了显著下调。低温胁迫下，差异表达基因显著富集到细胞凋亡、直肠癌和癌症的转录误调节通路，说明细胞的增殖和凋亡受到影响。显著变化的lncRNA主要以反式调控作用于靶基因，且与靶基因间的互作关系复杂，它们调控的靶基因富集到单纯疱疹病毒1感染通路、MAPK信号通路和范可尼贫血通路。此外，再无其他通路受到影响。低温胁迫影响了民猪的神经和免疫系统，使细胞内的物质转运发生了变化，细胞的生长、分化也受到了影响，但由于基因发生变化的倍数较小（1～2倍为主），且受影响的通路较少（仅3条），因此推测低温胁迫并未对民猪的骨骼肌造成严重损伤。

关键词: 民猪, 骨骼肌, 低温胁迫, RNA-seq, lncRNA

Abstract: The study aimed to explore the genetic mechanism of Min pigs excellent germplasm characteristics, and analyze their low temperature adaptation mechanism.In this experiment, 6 female Min pigs at the age of 6 months with similar body weight were randomly divided into 2 groups with 3 individuals in each group. Individuals in the control group were raised in a pig house whose temperature was controlled at (18±2)℃, and individuals in the experimental group were raised in a semi-open house outside. The ambient temperature dropped from 5℃/-5℃ on the first day to -15℃/-24℃ on the last day. The experiment lasted 58 days. Both groups of individuals were guaranteed free access to food and water. At the end of the experiment, all individuals were slaughtered, and the longissimus dorsi was taken for RNA-seq. The genes and lncRNAs induced by low temperature in Min pig skeletal muscle were screened, and their functions were annotated and their regulatory relationships were analyzed. The RNA sequencing analysis showed that 86 genes in skeletal muscle were significantly up-regulated and 16 genes were significantly down-regulated after 58 days of low temperature stress, 112 lncRNAs were significantly up-regulated and 74 lncRNAs were significantly down-regulated. Among the genes that were significantly up-regulated, 4 nervous system-related genes NTSR2, ARC, FOSL1 and RCAN1 were significantly up-regulated, and 7 solute transport related genes SLC2A4, SLC2A5, SLC4A10, SLC19A2, SLC20A1, SLC28A1 and SLC38A2, 6 genes AREG, CISH, OTUD1, TRIB1, GPA33 and ITPKC related to inflammation and immunity were significantly up-regulated. The circadian rhythm-related ARNTL gene and the RASL11A gene inhibiting cell proliferation, were significantly down-regulated. Under low temperature stress, differentially expressed genes were significantly enriched in apoptosis pathway, colorectal cancer pathway and transcriptional misregu-lation in cancer pathway, indicating that cell proliferation and apoptosis were affected. Significantly changed lncRNAs mainly regulated target genes in trans, and have complex interactions with target genes. The target genes regulated by lncRNAs were enriched in Herpes simplex virus 1 infection pathway, MAPK signaling pathway and Fanconi anemia pathway. In addition, no other pathways were affected. Low temperature stress affects the nervous and immune systems of Min pigs, changes the transport of substances in the cells, and affects the growth and differentiation of cells. However, because the fold of gene changes is small (mainly 1-2 times) and fewer pathways (only 3) are affected, it is speculated that low temperature stress does not cause serious damage to the skeletal muscle of Min pig.

Key words: Min pig, skeletal muscle, low temperature stress, RNA-seq, lncRNA

中图分类号:

S828.2

张冬杰, 汪亮, 马红, 李忠秋, 王文涛, 刘娣. 低温胁迫下民猪骨骼肌的转录调控分析[J]. 畜牧兽医学报, 2022, 53(8): 2524-2536.

ZHANG Dongjie, WANG Liang, MA Hong, LI Zhongqiu, WANG Wentao, LIU Di. Analysis of Transcriptional Regulation in Min Pig Skeletal Muscle under Low Temperature Stress[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2524-2536.

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