犬弓首蛔虫感染比格犬不同阶段肝circRNAs表达模式的分析

doi:10.11843/j.issn.0366-6964.2021.012.019

Abstract

Abstract: This study was conducted to investigate the potential regulatory role of circRNAs in the definitive host liver in the pathogenesis induced by Toxocara canis. The six-to-seven-week-old Beagle puppies (n=18) were averagely divided into three stages (0.5 day post infection, 1 dpi, and 36 dpi), and their liver samples were collected at the corresponding time points after infection. Each stage included infected group and control group. Total RNA was extracted from the livers of infected and uninfected puppies, and the circRNA libraries were constructed and sequenced by high-throughput RNA sequencing. The differentially expressed (DE) circRNAs were analyzed, and GO and KEGG enrichment analysis were performed using the source genes of DE circRNAs. The nine DEcircRNAs were randomly selected for quantitative real-time PCR (RT-qPCR) verification. Compared to the control groups, 94, 103, and 84 DE circRNAs of dogs' livers were detected in the livers of infected dogs at the three infection stages, respectively. Venn plot analysis showed that there were no co-expressed DE circRNAs among the three stages of infection. GO and KEGG analysis showed that some DE circRNAs were related to immunity and inflammation in the host liver. Among these DE circRNAs, novel_circ_0016108, novel_circ_0016184 and novel_circ_0027468 were related to the natural immunity of host against T. canis infection; novel_circ_0002212 was involved in liver inflammation caused by T. canis; novel_circ_0002212 and novel_circ_0019907 play a role in the host liver's resistance to T. canis invasion. RT-qPCR validation showed that the expression trends of DE circRNAs obtained by both methods were consistent. These results suggest that circRNAs play an important role in the infection of definitive host (dogs) by T. canis. This study provided new information for further understanding of the interaction between T. canis and its definitive host, and will also facilitate the development of interventions for the disease.

Key words: Toxocara canis, Beagle dogs, liver, RNA sequencing, circRNAs

CLC Number:

S855.9

ZOU Yang, ZHENG Wenbin, ZHANG Jinpeng, LU Yixin, ZHU Xingquan. CircRNAs Expression Patterns in the Liver of Beagle Dogs at Different Stages of Toxocara canis Infection[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3524-3534.

References 41

[1]	WINDERS W T, MENKIN-SMITH L. Toxocara canis[M]//StatPearls. Treasure Island:StatPearls Publishing, 2020.
[2]	CONG W, ZHANG X X, ZHOU N, et al. Toxocara seroprevalence among clinically healthy individuals, pregnant women and psychiatric patients and associated risk factors in Shandong Province, Eastern China[J]. PLoS Negl Trop Dis, 2014, 8(8):e3082.
[3]	CONG W, MENG Q F, YOU H L, et al. Seroprevalence and risk factors of Toxocara infection among children in Shandong and Jilin provinces, China[J]. Acta Trop, 2015, 152:215-219.
[4]	YANG G L, ZHANG X X, SHI C W, et al. Seroprevalence and associated risk factors of Toxocara infection in Korean, Manchu, Mongol, and Han ethnic groups in northern China[J]. Epidemiol Infect, 2016, 144(14):3101-3107.
[5]	DESPOMMIER D. Toxocariasis:clinical aspects, epidemiology, medical ecology, and molecular aspects[J]. Clin Microbiol Rev, 2003, 16(2):265-272.
[6]	MA G X, HOLLAND C V, WANG T, et al. Human toxocariasis[J]. Lancet Infect Dis, 2018, 18(1):E14-E24.
[7]	MOREIRA G M S G, DE LIMA TELMO P L, MENDONÇA M, et al. Human toxocariasis:current advances in diagnostics, treatment, and interventions[J]. Trends Parasitol, 2014, 30(9):456-464.
[8]	HAYASHI E, TUDA J, IMADA M, et al. The high prevalence of asymptomatic Toxocara infection among schoolchildren in Manado, Indonesia[J]. Southeast Asian J Trop Med Public Health, 2005, 36(6):1399-1406.
[9]	FU C J, CHUANG T W, LIN H S, et al. Seroepidemiology of Toxocara canis infection among primary schoolchildren in the capital area of the Republic of the Marshall Islands[J]. BMC Infect Dis, 2014, 14(1):261.
[10]	DDAWELA R D, RAJAPAKSE R P V J, PERERA N A N D, et al. Characterization of a Toxocara canis species-specific excretory-secretory antigen (TcES-57) and development of a double sandwich ELISA for diagnosis of visceral larva migrans[J]. Korean J Parasitol, 2007, 45(1):19-26.
[11]	ZHENG W B, ZOU Y, HE J J, et al. Global profiling of lncRNAs-miRNAs-mRNAs reveals differential expression of coding genes and non-coding RNAs in the lung of beagle dogs at different stages of Toxocara canis infection[J]. Int J Parasitol, 2021, 51(1):49-61.
[12]	ZOU Y, ZHENG W B, HE J J, et al. Toxocara canis differentially affects hepatic microRNA expression in Beagle Dogs at different stages of infection[J]. Front Vet Sci, 2020, 7:587273.
[13]	CONN S J, PILLMAN K A, TOUBIA J, et al. The RNA binding protein quaking regulates formation of circRNAs[J]. Cell, 2015, 160(6):1125-1134.
[14]	GRANADOS-RIVERON J T, AQUINO-JARQUIN G. The complexity of the translation ability of circRNAs[J]. Biochim Biophys Acta, 2016, 1859(10):1245-1251.
[15]	ZHENG W B, ZOU Y, ELSHEIKHA H M, et al. Serum metabolomic alterations in Beagle dogs experimentally infected with Toxocara canis[J]. Parasit Vectors, 2019, 12(1):447.
[16]	REGIS S C S, MENDONCA L R, DOS SANTOS SILVA N, et al. Seroprevalence and risk factors for canine toxocariasis by detection of specific IgG as a marker of infection in dogs from salvador, Brazil[J]. Acta Trop, 2011, 120(1-2):46-51.
[17]	SONG F B, WANG L M, ZHU W B, et al. Long noncoding RNA and mRNA expression profiles following igf3 knockdown in common carp, Cyprinus carpio[J]. Sci Data, 2019, 6:190024.
[18]	ZHOU J, XIONG Q M, CHEN H T, et al. Identification of the spinal expression profile of non-coding RNAs involved in neuropathic pain following spared nerve injury by sequence analysis[J]. Front Mol Neurosci, 2017, 10:91.
[19]	LANGMEAD B, SALZBERG S L. Fast gapped-read alignment with bowtie 2[J]. Nat Methods, 2012, 9(4):357-359.
[20]	MEMCZAK S, JENS M, ELEFSINIOTI A, et al. Circular RNAs are a large class of animal RNAs with regulatory potency[J]. Nature, 2013, 495(7441):333-338.
[21]	GAO Y, WANG J F, ZHAO F Q. CIRI:an efficient and unbiased algorithm for de novo circular RNA identification[J]. Genome Biol, 2015, 16(1):4.
[22]	ZHOU L, CHEN J H, LI Z Z, et al. Integrated profiling of microRNAs and mRNAs:microRNAs located on Xq27. 3 associate with clear cell renal cell carcinoma[J]. PLoS One, 2010, 5(12):e15224.
[23]	TAMURA K, NEI M. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees[J]. Mol Biol Evol, 1993, 10(3):512-526.
[24]	KUMAR S, STECHER G, TAMURA K. MEGA7:molecular evolutionary genetics analysis version 7. 0 for bigger datasets[J]. Mol Biol Evol, 2016, 33(7):1870-1874.
[25]	FELSENSTEIN J. Confidence limits on phylogenies:an approach using the bootstrap[J]. Evolution, 1985, 39(4):783-791.
[26]	SHANGGUAN W, LIANG X, SHI W, et al. Identification and characterization of circular RNAs in rapid atrial pacing dog atrial tissue[J]. Biochem Biophys Res Commun, 2018, 506(1):1-6.
[27]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the method[J]. Methods, 2001, 25(4):402-408.
[28]	YOUNG M D, WAKEFIELD M J, SMYTH G K, et al. Gene ontology analysis for RNA-seq:accounting for selection bias[J]. Genome Biol, 2010, 11(2):R14.
[29]	KANEHISA M, ARAKI M, GOTO S, et al. KEGG for linking genomes to life and the environment[J]. Nucleic Acids Res, 2008, 3(S1)6:D480-D484.
[30]	卜卜才加. 犬弓首蛔虫烯醇化酶基因的克隆、序列分析及免疫效果评价[D]. 长春:吉林农业大学, 2018.BUBU C J. Cloning, sequence analysis and immune evaluation of Toxocara canis enolase gene[D]. Changchun:Jilin Agricultural University, 2018. (in Chinese)
[31]	KRISTENSEN L S, ANDERSEN M S, STAGSTED L V W, et al. The biogenesis, biology and characterization of circular RNAs[J]. Nat Rev Genet, 2019, 20(11):675-691.
[32]	SCHNIEDER T, LAABS E M, WELZ C. Larval development of Toxocara canis in dogs[J]. Vet Parasitol, 2011, 175(3-4):193-206.
[33]	WIPKE B T, ALLEN P M. Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis[J]. J Immunol, 2001, 167(3):1601-1608.
[34]	MONACH P A, NIGROVIC P A, CHEN M, et al. Neutrophils in a mouse model of autoantibody-mediated arthritis:critical producers of Fc receptor γ, the receptor for C5a, and lymphocyte function-associated antigen 1[J]. Arthritis Rheum, 2010, 62(3):753-764.
[35]	NÉMETH T, MÓCSAI A, LOWELL C A. Neutrophils in animal models of autoimmune disease[J]. Semin Immunol, 2016, 28(2):174-186.
[36]	LIU Z, GIUDICE G J, ZHOU X, et al. A major role for neutrophils in experimental bullous pemphigoid[J]. J Clin Invest, 1997, 100(5):1256-1263.
[37]	CHAN S T, OU J H J. Hepatitis C virus-induced autophagy and host innate immune response[J]. Viruses, 2017, 9(8):224.
[38]	RATH M, MVLLER I, KROPF P, et al. Metabolism via Arginase or nitric oxide synthase:two competing arginine pathways in macrophages[J]. Front Immunol, 2014, 5:532.
[39]	RIBEIRO-GOMES F L, ROMA E H, CARNEIRO M B, et al. Site-dependent recruitment of inflammatory cells determines the effective dose of Leishmania major[J]. Infect Immun, 2014, 82(7):2713-2727.
[40]	ULLAND A E, SHEARER J D, COULTER C, et al. Altered wound arginine metabolism by corticosterone and retinoic acid[J]. J Surg Res, 1997, 70(1):84-88.
[41]	SUZUKI T, ICHII O, NAKAMURA T, et al. Immune-associated renal disease found in caspase 3-deficient mice[J]. Cell Tissue Res, 2020, 379(2):323-335.

CircRNAs Expression Patterns in the Liver of Beagle Dogs at Different Stages of Toxocara canis Infection

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