Abstract:Long-chain non-coding RNA (lncRNA) has become an important participant in almost all gene functions and regulatory levels. In order to reveal the potential effect of lncRNA on the milk yield of horses (Equus caballus) in early lactation, 8 healthy Kazakh horses of 3~4 years old with similar body shape and early lactation were selected in this study, and they were divided into high milk yield group (H) and low milk yield group (L) according to their milk yield. The non-coding RNA in the milk of 2 groups of horses was sequenced by transcriptome sequencing technology. Through the significant enrichment analysis of GO and Pathway functions, the main biological functions of differentially expressed genes, biochemical metabolic pathways and signal pathways involved were determined. According to P<0.05 and FC≥1, the differentially expressed lncRNA was screened out, and the function of the differentially expressed lncRNA was further studied. The results showed that a total of 30 differentially expressed lncRNA (up-regulated 16, down-regulated 14) were found in Kazakh horse milk of high-yield group and low-yield group in early lactation. Genes related to breast development and lactation, such as vascular endothlial growth factor, (VEGFD), phosphatidylinositol glycan, class A (PIGA), and aconitase closing odor (ACO1) were screened out. The results showed that lncRNA may regulate the process of lactation or mammary gland development by targeting genes related to mammary gland development and lactation. This study provides reference for the application of lncRNA in horse breeding.
马应国, 梁金超, 马海玉, 刘玲玲, 刘武军. 基于转录组测序技术对哈萨克马产奶量相关差异表达 lncRNA 的研究[J]. 农业生物技术学报, 2023, 31(5): 1001-1012.
MA Ying-Guo, LIANG Jin-Chao, MA Hai-Yu, LIU Ling-Ling, LIU Wu-Jun. Study of Differentially Expressed LncRNA Related to Milk Production in Kazakh Horses (Equus caballus) Based on Transcriptome Sequencing Technology. 农业生物技术学报, 2023, 31(5): 1001-1012.
[1] 崔英俊, 于广璞, 汤云飞. 2018. FGF2 通过 MAPK 信号通路影响奶牛乳腺分支形态变化[J]. 东北农业大学学报, 49(08): 48-53, 72. (Cui Y J, Yu G P, Tang Y F. 2018. FGF2 affects the morphological changes of mammary branches of dairy cows through MAPK signaling pathway[J]. Journal of Northeast Agricultural University, 49(08): 48-53, 72.) [2] 杜晓军, 周娟. 2007. 松弛素在心血管疾病中的作用及临床治疗展望[J]. 西安交通大学学报( 医学版),(02): 117-122, 125.122, 125.) [3] 郝志云. 2021. 绵羊泌乳性状非编码 RNA 筛选、鉴定及功能研究[D]. 博士学位论文, 甘肃农业大学, 导师: 罗玉柱, 王继卿. pp. 19-64. (Hao Z Y. 2021. Screening, Identification and function study of non-coding RNA for lacta tion traits of sheep[D]. Thesis for Ph. D. Gansu Agricultural University, Supervisor: Luo Y Z, Wang J Q. pp. 19-64.) [4] 侯晓明. 2009. 奶牛乳腺发育及泌乳重要功能基因的筛选[D]. 博士学位论文, 东北农业大学, 导师: 李庆章. pp.13-18. (Hou X M. 2009. Screening of important functional genes for mammary gland development and lactation in dairy cows[D]. Thesis for Ph. D. Northeast Agricultural University, Supervisor: Li Q Z, pp. 13-18.) [5] 侯永清, 姚康, 印遇龙, 等. 2016. 氨基酸的内源性合成限制了动物的生长, 泌乳和繁殖 ( Ⅱ)[J]. 中国饲料(10): 7 (Hou Y Q, Yao K, Yin Y L, et al. 2016. Endogenous synthesis of amino acids restricts animal growth, lactation and reproduction (10): 7.) [6] 姜勇, 罗深秋. 2005. 细胞信号转导的分子基础与功能调控 [M]. 科学出版社, 北京. pp. 141-146. (Jiang Y, Luo S Q. 2005. Molecular basis and functional regulation of cell signal transduction[M]. Science Press, BeiJing. pp. 141-146.) [7] 牛春阳, 薛琳琳, 计红, 等. 2019. Lncrna 生物学功能研究进展[J]. 中国生物制品学杂志,(2): 6. (Niu C Y, Xue L L, Ji H, et al. 2019. Research progress on biological functions of LncRNA[J]. Chinese Journal of Biologicals, (2): 6.) [8] 宿娅, 张晨芳, 魏强, 等. 2014. 长链非编码 RNA 研究进展[J]. 西北植物报, 34(11): 2357-2365. (Su Y, Zhang C F, Wei Q, et al. 2014. Research progress of long-chain noncoding RNA[J]. Journal of Northwest Botany, 34(11): 2357-2365.) [9] 余元勋. 2013. 中国疾病信号通路与靶向治疗学[M]. 安徽科学技术出版社.. pp. 208-229. (Yu Y X. 2013. Chinese disease signaling pathway and targeted therapy[M]. Anhui Science and Technology Press.. pp. 208-229.) [10] 王鲲. 2017. 血管紧张素转化酶 2 在 LPS 诱导的小鼠乳腺炎症损伤中的保护作用及其分子机制[D]. 博士学位论文, 南京农业大学, 导师: 张源淑. pp. 97-106. (. Wang K. 2017. Protective effect of angiotensin-converting enzyme 2 on LPS-induced inflammation of mouse mammary gland and its molecular mechanism[D]. Thesis for Ph. D., Nanjing Agricultural University, Supervisor: Zhang Y S. pp. 97-106.) [11] 杨兵. 2019. 奶牛乳腺差异表达长链非编码 RNA 的筛选、鉴定及其功能研究[D]. 博士学位论文, 西北农林科技大学, 导师: 王昕, pp. 67-74. (Yang B. 2019. Screening, identification and functional study of long-chain non- coding RNA differentially expressed in milk cow mammary gland[D]. Thesis for Ph. D., Northwest A&F University, Supervisor: Wang X, pp. 67-74.) [12] 赵行宇, 吕士杰, 沈楠. 2005. RAS 蛋白的信号转导途径[J]. 吉林医药学院学报, (03): 169-171. (Zhao X Y, Lu S J, Shen N. 2005. Signal transduction pathway of RAS protein[J]. Journal of Jilin Medical College, (03): 169-171.) [13] Akers R M. 2017. A 100-Year Review: Mammary development and lactation[J]. Journal of Dairy Science, 100(12): 10332-10352. [14] Birney E, Stamatoyannopoulos J A, Dutta A, et al. 2007. Identification and analysis of functional elements in 1% of the human genome by the encode pilot project[J]. Nature, 447: 799-816. [15] Böhmdorfer G, Wierzbicki A T. 2015. Control of chromatin structure by long noncoding RNA[J]. Trends in Cell Biology, 25(10):623-632. [16] Burgos S A, Cant J P. 2010. IGF-1 stimulates protein synthesis by enhanced signaling through mTORC1 in bovine mammary epithelial cells[J]. Domestic Animal Endocrinology, 38(4): 211-221. [17] Datta S R, Brunet A, Greenberg M E. 1999. Cellular survival: A play in three Akts[J]. Genes Development, 13(22): 2905-2927. [18] Diaz-Torga G, Feierstein C, Libertun C, et al. 2002. Disruption of the D2 dopamine receptor alters GH and IGF-1 secretion and causes dwarfism in male mice[J]. Endocrinology, 143(4): 1270-1279. [19] Garnsworthy P C, Masson L L, Lock A L, et al. 2006. Variation of milk citrate with stage of lactation and de novo fatty acid synthesis in dairy cows[J]. Journal of Dairy Science, 89(5): 1604-1612. [20] De Groot N, Van Kuik-Romeijn P, Lee S H, et al. 2000. Increased immunoglobulin a levels in milk by over-expressing the murine polymeric immunoglobulin receptor gene in the mammary gland epithelial cells of transgenic mice[J]. Immunology, 101(2): 218-224. [21] Hung T, Wang Y, Lin M F, et al. 2011. Extensive and coordinated transcription of noncoding rnas within cell-cycle promoters[J]. Nature Genetics, 43(7): 621-629. [22] Ibeagha-Awemu E M, Li R, Dudemaine P L, et al. 2018. Transcriptome analysis of long non-coding RNA in the bovine mammary gland following dietary supplementa tion with linseed oil and safflower oil[J]. International Journal of Molecular Sciences, 19(11): 3610-3631. [23] Ji Z, Chao T, Liu Z, et al. 2020. Genome-wide integrated analysis demonstrates widespread functions of lncRNAs in mammary gland development and lactation in dairy goats[J]. BMC Genomics, 21: 1-11. [24] Kino M T, Hur D E, Ichijo T, et al. 2010. Noncoding rna gas5 is a growth arrest and starvation- associated repressor of the glucocorticoid receptor[J]. Science Signalling, 3 (107): ra8. [25] Knowling S, Morris K V. 2011. Non-coding rna and antisense rna. nature's trash or treasure?[J]. Biochimie, 93(11): 1922-1927. [26] Lu Y, Liu X, Xie M, et al. 2017. The NF- κB-responsive long noncoding RNAfirre regulates posttranscriptional regulation of inflammatory gene expression through interacting with hnRNPU[J]. The Journal of Immunology, 199(10): 3571-3582. [27] Mohammad M A, Haymond M W. 2013. Regulation of lipid synthesis genes and milk fat production in human mammary epithelial cells during secretory activation[J]. American Journal of Physiology. Endocrinology and Metabolism, 305(6): E700-E716. [28] Musaro A, Giacinti C, Borsellino G, et al. 2004. Stem cell mediated muscle regeneration is enhance by local isoform of insulin like growth factor 1[J]. Proceedings of the National Academy of Sciences of USA, 101(5): 1206-1210. [29] Ni Y F, Chen F, Cai Q Q, et al. 2020. Long noncoding rna and mrna profiling of hypothalamic-pituitary-mammary gland axis in lactating sows under heat stress[J]. Genomics, 112(5): 3668-3676. [30] Nishikawa K, Kinjo A R. 2017. Essential role of long non-coding rnas in de novo chromatin modifications: The genomic address code hypothesis[J]. Biophysical Reviews, 9: 73-77. [31] Orom U A, Derrien T, Beringer M, et al. 2010. Long noncoding rnas with enhancer-like function in human cells[J]. Cell, 143: 46-58 [32] Presneau N, Shalaby A, Idowu B, et al. 2009. Potential therapeutic targets for chordoma: PI3K/AKT/TSC1/TSC2/ mTOR pathway[J]. British Joural of Cancer, 100(9): 1406-1414. [33] Raynal L K, Park Y W, Gaucheron F, et al. 2017. Heat stability and enzymatic modifications of goat and sheep milk[J]. Small Ruminant Research, 68(1-2): 207-220. [34] Rincheval A A, Belair L, Djiane J. 2002. Developmental expression of pigr gene in sheep mammary gland and hormonal regulation[J]. Journal of Dairy Research, 69(1): 13-26. [35] Rinn J L, Chang H Y. 2012. Genome regulation by long noncoding RNAs[J]. Annual Review of Biochemistry, 81: 145-166. [36] Rosato R, Jammes H, Belair L, et al. 1995. Polymericig receptor gene expression in rabbit mammary gland during pregnancy and lactation: Evolution and hormonal regulation[J]. Molecular & Cellular Endocrinology, 110(1): 81. [37] Rosen C J, Pollak M, et al. 1999. Circulating Igf-1: New perpectives for a new century[J]. Trends in Endocrinology Metabolism, 10(4): 136-141. [38] Standaert L, Adriaens C, Radaelli E, et al. 2014. The long noncoding rna neat1 is required for mammary gland development and lactation[J]. RNA, 20(12): 1844-1849. [39] Twigg S M, Baxter R C. 1998. IGF binding protein 5 forms an alternative ternary complex with IGFs and the acid lable subunit[J]. Journal of Biological Chemistry, 273(11): 6074-6079. [40] Wang X X, Wang H, Zhang R Q, et al. 2020. LRRC75A antisense lncRNA1 knockout attenuates inflammatory responses of bovine mammary epithelial cells[J]. International Journal of Biological Sciences, 16(2): 251-263. [41] Yang B, Jiao B, Ge W, et al. 2018. Transcriptome sequencing to detect the potential role of long non-coding RNAs in bovine mammary gland during the dry and lactation period[J]. BMC Genomics, 19(1): 605. [42] Yu S, Zhao Y, Lai F, et al. 2017. Lncrna as cernas may be involved in lactation process[J]. Oncotarget, 8(58): 98014-98028. [43] Zhao L, Roche P J, Gunnersen J M, et al. 1999. Mice without a functional relaxin gene are unable to deliver milk to their pups[J]. Endorcinology, 140: 445-453. [44] Zheng X, Ning C, Zhao P, et al. 2018. Integrated analysis of long noncoding RNA and mRNA expression profiles reveals the potential role of long noncoding RNA in different bovine lactation stages[J]. Journal of Dairy Science, 101(12): 11061-11073