Distribution and differential expression of microRNAs in the intestinal mucosal layer of necrotic enteritis induced Fayoumi chickens
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Objective: Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer. Methods: NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction. Results: According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, gga-miR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, gga-miR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other. Conclusion: Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.
Abstract
Objective: Despite an increasing number of investigations into the pathophysiology of necrotic enteritis (NE) disease, etiology of NE-associated diseases, and gene expression profiling of NE-affected tissues, the microRNA (miRNA) profiles of NE-affected poultry have been poorly studied. The aim of this study was to induce NE disease in the genetically disparate Fayoumi chicken lines, and to perform non-coding RNA sequencing in the intestinal mucosal layer. Methods: NE disease was induced in the Fayoumi chicken lines (M5.1 and M15.2), and non-coding RNA sequencing was performed in the intestinal mucosal layer of both NE-affected and uninfected chickens to examine the differential expression of miRNAs. Next, quantitative real-time polymerase chain reaction (real-time qPCR) was performed to further examine four miRNAs that showed the highest fold differences. Finally, bioinformatics analyses were performed to examine the four miRNAs target genes involvement in the signaling pathways, and to examine their interaction. Results: According to non-coding RNA sequencing, total 50 upregulated miRNAs and 26 downregulated miRNAs were detected in the NE-induced M5.1 chickens. While 32 upregulated miRNAs and 11 downregulated miRNAs were detected in the NE-induced M15.2 chickens. Results of real-time qPCR analysis on the four miRNAs (gga-miR-9-5p, gga-miR-20b-5p, gga-miR-196-5p, and gga-let-7d) were mostly correlated with the results of RNAseq. Overall, gga-miR-20b-5p was significantly downregulated in the NE-induced M5.1 chickens and this was associated with the upregulation of its top-ranking target gene, mitogen-activated protein kinase, kinase 2. Further bioinformatics analyses revealed that 45 of the gene targets of gga-miR-20b-5p were involved in signal transduction and immune system-related pathways, and 35 of these targets were predicted to interact with each other. Conclusion: Our study is a novel report of miRNA expression in Fayoumi chickens, and could be very useful in understanding the role of differentially expressed miRNAs in a NE disease model.
참고문헌 (30)
- Timbermont L, "Necrotic enteritis in broilers: an updated review on the pathogenesis", Avian Pathol, 40, pp.341~347 (2011)
- Broussard CT, "Necrotic enteritis in cage-reared commercial layer pullets", Avian Dis, 30, pp.617~619 (1986)
- Jang SI, "Vaccination with Clostridium perfringens recombinant proteins in combination with Montanide ISA 71 VG adjuvant increases protection against experimental necrotic enteritis in commercial broiler chickens", Vaccine, 30, pp.5401~5406 (2012)
- Dahiya JP, "Potential strategies for controlling necrotic enteritis in broiler chickens in post-antibiotic era", Anim Feed Sci Technol, 129, pp.60~88 (2006)
- Oh ST, "The role of host genetic factors and host immunity in necrotic enteritis", Avian Pathol, 45, pp.313~316 (2016)
- Sun W, "microRNA: A master regulator of cellular processes for bioengineering systems", Annu Rev Biomed Eng, 12, pp.1~27 (2010)
- Rengaraj D, "Regulation of glucose phosphate isomerase by the 3'UTR-specific miRNAs miR-302b and miR-17-5p in chicken primordial germ cells", Biol Reprod, 89, pp.33~ (2013)
- Croce CM, "Causes and consequences of microRNA dysregulation in cancer", Nat Rev Genet, 10, pp.704~714 (2009)
- Dinh H, "Modulation of microRNAs in two genetically disparate chicken lines showing different necrotic enteritis disease susceptibility", Vet Immunol Immunopathol, 159, pp.74~82 (2014)
- Hong YH, "Differential regulation of microRNA transcriptome in chicken lines resistant and susceptible to necrotic enteritis disease", Poult Sci, 93, pp.1383~1395 (2014)
- Pinard-Van Der Laan MH, "Comparison of outbred lines of chickens for resistance to experimental infection with coccidiosis (Eimeria tenella)", Poult Sci, 77, pp.185~191 (1998)
- Kim DK, "Genetically disparate Fayoumi chicken lines show different response to avian necrotic enteritis", J Poult Sci, 52, pp.245~252 (2015)
- Zhou H, "Genetic characterization of biodiversity in highly inbred chicken lines by microsatellite markers", Anim Genet, 30, pp.256~264 (1999)
- Kim DK, "Gene expression profiles of two B-complex disparate, genetically inbred Fayoumi chicken lines that differ in susceptibility to Eimeria maxima", Poult Sci, 88, pp.1565~1579 (2009)
- Kozomara A, "miRBase: annotating high confidence microRNAs using deep sequencing data", Nucleic Acids Res, 42, pp.D68~D73 (2014)
- Saeed AI, "TM4: a free, open-source system for microarray data management and analysis", Biotechniques, 34, pp.374~378 (2003)
- Wong N, "miRDB: an online resource for microRNA target prediction and functional annotations", Nucleic Acids Res, 43, pp.D146~D152 (2015)
- Kanehisa M, "KEGG as a reference resource for gene and protein annotation", Nucleic Acids Res, 44, pp.D457~D462 (2016)
- Szklarczyk D, "STRING v10: protein-protein interaction networks, integrated over the tree of life", Nucleic Acids Res, 43, pp.D447~D452 (2015)
- Bartel DP, "MicroRNAs: genomics, biogenesis, mechanism, and function", Cell, 116, pp.281~297 (2004)
- Ambros V, "A uniform system for microRNA annotation", RNA, 9, pp.277~279 (2003)
- Luo W, "E2F1-miR-20a-5p", 20b-5p auto-regulatory feedback loop involved in myoblast proliferation and differentiation, 2016, pp.6, 27904~
- Yoshimura K, "Immunolocalization of aquaporin-4 in the brain, kidney, skeletal muscle, and gastro-intestinal tract of chicken", Cell Tissue Res, 344, pp.51~61 (2011)
- Winsauer G, "XIAP regulates bi-phasic NF-kappaB induction involving physical interaction and ubiquitination of MEKK2", Cell Signal, 20, pp.2107~2112 (2008)
- He Y, "MAP3K3 expression in tumor cells and tumor-infiltrating lymphocytes is correlated with favorable patient survival in lung cancer", Sci Rep, 5, pp.11471~ (2015)
- Li L, "Proinflammatory role of aquaporin-4 in autoimmune neuroinflammation", FASEB J, 25, pp.1556~1566 (2011)
- Schmiedel D, "The RNA binding protein IMP3 facilitates tumor immune escape by downregulating the stress-induced ligands ULPB2 and MICB", Elife, 5 (2016)
- Shaughnessy RG, "Innate immune gene expression differentiates the early avian intestinal response between Salmonella and Campylobacter", Vet Immunol Immunopathol, 132, pp.191~198 (2009)
- Anh Duc Truong, "RNA-seq Profiles of Immune Related Genes in the Spleen of Necrotic Enteritis-afflicted Chicken Lines", ASIAN-AUSTRALASIAN JOURNAL OF ANIMAL SCIENCES, 28(10), pp.1496~1511 (2015), 아세아·태평양축산학회[DOI]
- Rengaraj D, "Expression analysis of cytosolic DNA-sensing pathway genes in the intestinal mucosal layer of necrotic enteritis-induced chicken", Vet Immunol Immunopathol, 170, pp.1~12 (2016)