Hostname: page-component-848d4c4894-75dct Total loading time: 0 Render date: 2024-05-24T22:43:53.325Z Has data issue: false hasContentIssue false

Expression profiles of miRNAs from bovine mammary glands in response to Streptococcus agalactiae-induced mastitis

Published online by Cambridge University Press:  23 August 2017

Junhua Pu
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Rui Li
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Chenglong Zhang
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Dan Chen
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Xiangxiang Liao
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Yihui Zhu
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Xiaohan Geng
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Dejun Ji
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Yongjiang Mao
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
Yunchen Gong
The Centre for the Analysis of Genome Evolution and Function (CAGEF), University of Toronto, Toronto, Canada
Zhangping Yang*
College of Animal Science and Technology, Yangzhou University, Yangzhou, China
*For correspondence; e-mail:


This study aimed to describe the expression profiles of microRNAs (miRNAs) from mammary gland tissues collected from dairy cows with Streptococcus agalactiae-induced mastitis and to identify differentially expressed miRNAs related to mastitis. The mammary glands of Chinese Holstein cows were challenged with Streptococcus agalactiae to induce mastitis. Small RNAs were isolated from the mammary tissues of the test and control groups and then sequenced using the Solexa sequencing technology to construct two small RNA libraries. Potential target genes of these differentially expressed miRNAs were predicted using the RNAhybrid software, and KEGG pathways associated with these genes were analysed. A total of 18 555 913 and 20 847 000 effective reads were obtained from the test and control groups, respectively. In total, 373 known and 399 novel miRNAs were detected in the test group, and 358 known and 232 novel miRNAs were uncovered in the control group. A total of 35 differentially expressed miRNAs were identified in the test group compared to the control group, including 10 up-regulated miRNAs and 25 down-regulated miRNAs. Of these miRNAs, miR-223 exhibited the highest degree of up-regulation with an approximately 3-fold increase in expression, whereas miR-26a exhibited the most decreased expression level (more than 2-fold). The RNAhybrid software predicted 18 801 genes as potential targets of these 35 miRNAs. Furthermore, several immune response and signal transduction pathways, including the RIG-I-like receptor signalling pathway, cytosolic DNA sensing pathway and Notch signal pathway, were enriched in these predicted targets. In summary, this study provided experimental evidence for the mechanism underlying the regulation of bovine mastitis by miRNAs and showed that miRNAs might be involved in signal pathways during S. agalactiae-induced mastitis.

Research Article
Copyright © Proprietors of Journal of Dairy Research 2017 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)


Audic, S & Claverie, JM 1997 The significance of digital gene expression profiles. Genome Research 7 986995 Google Scholar
Bartel, DP 2004 MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116 281297 Google Scholar
Bizuayehu, TT, Fernandes, JM, Johansen, SD & Babiak, I 2013 Characterization of novel precursor miRNAs using next generation sequencing and prediction of miRNA targets in Atlantic halibut. PLoS ONE 8 e61378 Google Scholar
Chao, WS 2008 Real-time PCR as a tool to study weed biology. Weed Science 56 290296 Google Scholar
Chen, L, Liu, X, Li, Z, Wang, H, Liu, Y, He, H, Yang, J, Niu, F, Wang, L & Guo, J 2014 Expression differences of miRNAs and genes on NF-κB pathway between the healthy and the mastitis Chinese Holstein cows. Gene 545 117125 CrossRefGoogle ScholarPubMed
Chen, YX, Gelfond, JAL, Mcmanus, LM & Shireman, PK 2009 Reproducibility of quantitative RT-PCR array in miRNA expression profiling and comparison with microarray analysis. BMC Genomics 10 407 Google Scholar
Dilda, F, Gioia, G, Pisani, LF, Restelli, L, Lecchi, C, Albonico, F, Bronzo, V, Mortarino, M & Ceciliani, F 2012 Escherichia coli lipopolysaccharides and Staphylococcus aureus enterotoxin B differentially modulate inflammatory microRNAs in bovine monocytes. Veterinary Journal 192 514516 Google Scholar
Dorn, GW 2011 MicroRNAs in cardiac disease. Translational Research the Journal of Laboratory and Clinical Medicine 157 226235 Google Scholar
Farnsworth, RJ 1987 Indications of contagious and environmental mastitis pathogens in a dairy herd. In Protocols 26th Annual Meeting National Mastitis Council, Orlando, FL, USA, Vol. 26, pp. 151155 Google Scholar
Fazi, F, Rosa, A, Fatica, A, Gelmetti, V, Marchis, MLD, Nervi, C & Bozzoni, I 2005 A minicircuitry comprised of microRNA-223 and transcription factors NFI-A and C/EBPα regulates human granulopoiesis. Cell 123 819831 Google Scholar
Filip, A 2007 MiRNA–new mechanisms of gene expression control. Postepy Biochemii 53 413419 Google ScholarPubMed
Franchini, M, Frattini, F, Crestani, S, Bonfanti, C & Lippi, G 2013 von Willebrand factor and cancer: a renewed interest. Thrombosis Research 131 290292 Google Scholar
Gantier, MP 2010 New perspectives in microRNA regulation of innate immunity. Journal of Interferon and Cytokine Research 30 283289 CrossRefGoogle ScholarPubMed
Gerin, I, Bommer, GT, McCoin, CS, Sousa, KM, Krishnan, V & MacDougald, OA 2010 Roles for miRNA-378/378* in adipocyte gene expression and lipogenesis. American Journal of Physiology Endocrinology and Metabolism 299 E198E206 CrossRefGoogle ScholarPubMed
Haneklaus, M, Gerlic, M, O'Neill, LA & Masters, SL 2013 miR-223: infection, inflammation and cancer. Journal of Internal Medicine 274 215226 Google Scholar
Heikkilä, AM, Nousiainen, JI & Pyörälä, S 2012 Costs of clinical mastitis with special reference to premature culling. Journal of Dairy Science 95 139150 Google Scholar
Iliopoulos, D, Drosatos, K, Hiyama, Y, Goldberg, IJ & Zannis, VI 2010 MicroRNA-370 controls the expression of microRNA-122 and Cpt1alpha and affects lipid metabolism. Journal of Lipid Research 51 15131523 Google Scholar
Jin, W, Ibeagha-Awemu, EM, Liang, G, Beaudoin, F, Zhao, X & Guan, LL 2014 Transcriptome microRNA profiling of bovine mammary epithelial cells challenged with Escherichia coli or Staphylococcus aureus bacteria reveals pathogen directed microRNA expression profiles. BMC Genomics 15 181 CrossRefGoogle ScholarPubMed
Jing, Q, Huang, S, Guth, S, Zarubin, T, Motoyama, A, Chen, JM, Padova, FD, Lin, SC, Gram, H & Han, JH 2005 Involvement of microRNA in AU-rich element-mediated mRNA instability. Cell 120 623634 Google Scholar
Johnnidis, JB, Harris, MH, Wheeler, RT, Stehling-Sun, S, Lam, MH, Kirak, O, Brummelkamp, TR, Fleming, MD & Camargo, FD 2008 Regulation of progenitor cell proliferation and granulocyte function by microRNA-223. Nature 451 11251129 CrossRefGoogle ScholarPubMed
Kanehisa, M, Araki, M, Goto, S, Hattori, M, Hirakawa, M, Itoh, M, Katayama, T, Kawashima, S, Okuda, S, Tokimatsu, T & Yamanishi, Y 2008 KEGG for linking genomes to life and the environment. Nucleic Acids Research 36 D480D484 CrossRefGoogle ScholarPubMed
Keefe, GP 1997 Streptococcus agalactiae mastitis: a review. Canadian Veterinary Journal 38 429437 Google Scholar
Li, R, Zhang, CL, Liao, XX, Chen, D, Wang, WQ, Zhu, YH, Geng, XH, Ji, DJ, Mao, YJ, Gong, YC & Yang, ZP 2015 Transcriptome microRNA profiling of bovine mammary glands infected with Staphylococcus aureus . International Journal of Molecular Sciences 16 49975013 Google Scholar
Livak, KJ & Schmittgen, TD 2001 Analysis of relative gene expression data using real-time quantitative PCR and the 2 −△△C T Method. Methods 25 402408 Google Scholar
Naeem, A, Zhong, K, Moisá, SJ, Drackley, JK, Moyes, KM & Loor, JJ 2012 Bioinformatics analysis of microRNA and putative target genes in bovine mammary tissue infected with Streptococcus uberis . Journal of Dairy Science 95 63976408 Google Scholar
O'Connell, RM, Rao, DS, Chaudhuri, AA & Baltimore, D 2010 Physiological and pathological roles for microRNAs in the immune system. Nature Reviews Immunology 10 111122 Google Scholar
Rinaldi, M, Li, RW & Capuco, AV 2010 Mastitis associated transcriptomic disruptions in cattle. Veterinary Immunology and Immunopathology 138 267279 Google Scholar
Sachdeva, M & Mo, YY 2010 MicroRNA-145 suppresses cell invasion and metastasis by directly targeting mucin 1. Cancer Research 70 378387 Google Scholar
Sordillo, LM & Streicher, KL 2002 Mammary gland immunity and mastitis susceptibility. Journal of Mammary Gland Biology Neoplasia 7 135146 Google Scholar
Starega-Roslan, J, Koscianska, E, Kozlowski, P & Krzyzosiak, WJ 2011 The role of the precursor structure in the biogenesis of microRNA. Cellular & Molecular Life Sciences 68 28592871 Google Scholar
Sun, J, Aswath, K, Schroeder, SG, Lippolis, JD, Reinhardt, TA & Sonstegard, TS 2015 MicroRNA expression profiles of bovine milk exosomes in response to Staphylococcus aureus infection. BMC Genomics 16 806815 Google Scholar
Taft, RJ, Pang, KC, Mercer, TR, Dinger, M & Mattick, JS 2010 Non-coding RNAs: regulators of disease. Journal of Pathology 220 126139 Google Scholar
Tang, G 2010 Plant microRNAs: an insight into their gene structures and evolution. Seminars in Cell and Developmental Biology 21 782789 CrossRefGoogle ScholarPubMed
Trigo, G, Ferreira, P, Ribeiro, N, Dinis, M, Andrade, EB, Melo-Cristino, J, Ramirez, M & Tavares, D 2008 Identification of immunoreactive extracellular proteins of Streptococcus agalactiae in bovine mastitis. Canadian Journal of Microbiology 54 899905 Google Scholar
Vimalraj, S & Selvamurugan, N 2013 MicroRNAs: synthesis, gene regulation and osteoblast differentiation. Current Issues in Molecular Biology 15 718 Google Scholar
Williams, AE, Perry, MM, Moschos, SA, Larner-Svensson, HM & Lindsay, MA 2008 Role of miRNA-146a in the regulation of the innate immune response and cancer. Biochemical Society Transactions 36 12111215 Google Scholar
Yan, M, Li, X, Tong, D, Han, C, Zhao, R, He, Y & Jin, X 2016 miR-136 suppresses tumor invasion and metastasis by targeting RASAL2 in triple-negative breast cancer. Oncology Reports 36 6571 Google Scholar
Yanaba, K, Asano, Y, Noda, S, Akamata, K, Aozasa, N, Taniguchi, T, Takahashi, T, Ichimura, Y, Toyama, T, Sumida, H, Kuwano, Y, Tada, Y, Sugaya, M, Kadono, T & Sato, S 2012 Augmented production of soluble CD93 in patients with systemic sclerosis and clinical association with severity of skin sclerosis. British Journal of Dermatology 167 542547 Google Scholar
Yang, Y, Wu, J, Guan, H, Cai, J, Fang, L, Li, J & Li, M 2012 MiR-136 promotes apoptosis of glioma cells by targeting AEG-1 and Bcl-2. FEBS Letters 586 36083612 Google Scholar
Zadoks, RN, Middleton, JR, McDougall, S, Katholm, J & Schukken, YH 2011 Molecular epidemiology of mastitis pathogens of dairy cattle and comparative relevance to humans. Journal of Mammary Gland Biology Neoplasia 16 357372 Google Scholar
Supplementary material: PDF

Pu et al supplementary material

Pu et al supplementary material

Download Pu et al supplementary material(PDF)
PDF 4.2 MB