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RNA-Seq-based transcriptomic profiling of primary interstitial cells of Cajal in response to bovine viral diarrhea virus infection

  • Shengnan Li
  • Xinyan Hu
  • Ruixin Tian
  • Yanting Guo
  • Junzhen Chen
  • Zhen Li
  • Xinyan Zhao
  • Ling Kuang
  • Duoliang Ran
  • Hongqiong Zhao
  • Xiaohong Zhang
  • Jinquan Wang
  • Lining Xia
  • Jianbo Yue
  • Gang YaoEmail author
  • Qiang FuEmail author
  • Huijun ShiEmail author
Original Article

Abstract

Infections with bovine viral diarrhea virus (BVDV) contribute significantly to health-related economic losses in the beef and dairy industries and are widespread throughout the world. Severe acute BVDV infection is characterized by a gastrointestinal (GI) inflammatory response. The mechanism of inflammatory lesions caused by BVDV remains unknown. The interstitial cells of Cajal (ICC) network plays a pivotal role as a pacemaker in the generation of electrical slow waves for GI motility, and it is crucial for the reception of regulatory inputs from the enteric nervous system. The present study investigated whether ICC were a good model for studying GI inflammatory lesions caused by BVDV infection. Primary ICC were isolated from the duodenum of Merino sheep. The presence of BVDV was detected in ICC grown for five passages after BVDV infection, indicating that BVDV successfully replicated in ICC. After infection with BVDV strain TC, the cell proliferation proceeded slowly or declined. Morphological changes, including swelling, dissolution, and formation of vacuoles in the ICC were observed, indicating quantitative, morphological and functional changes in the cells. RNA sequencing (RNA-Seq) was performed to investigate differentially expressed genes (DEGs) in BVDV-infected ICC and explore the molecular mechanism of underlying quantitative, morphological and functional changes of ICC. Eight hundred six genes were differentially expressed after BVDV infection, of which 538 genes were upregulated and 268 genes were downregulated. Gene Ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that the 806 DEGs were significantly enriched in 27 pathways, including cytokine-cytokine receptor interaction, interleukin (IL)-17 signaling and mitogen-activated protein kinase (MAPK) signaling pathways. The DEGs and raw files of high-throughput sequencing of this study were submitted to the NCBI Gene Expression Omnibus (GEO) database (accession number GSE122344). Finally, 21 DEGs were randomly selected, and the relative repression levels of these genes were tested using the quantitative real-time PCR (qRT-PCR) to validate the RNA-Seq results. The results showed that the related expression levels of 21 DEGs were similar to RNA-Seq. This study is the first to establish a new infection model for investigating GI inflammatory lesions induced by BVDV infection. RNA-Seq-based transcriptomic profiling can provide a basis for study on BVDV-associated inflammatory lesions.

Keywords

Bovine viral diarrhea virus Interstitial cells of Cajal RNA-Seq-based transcriptome profiling 

Notes

Acknowledgments

This work was supported by the Natural Science Foundation of China (Grant No. 31760742, 31502095 and 31560328), Key research and development plan of of Xinjiang Uyghur Autonomous Region (Grant No. 2017B01001-2), Postdoctoral Research Funds of China (Grant No. 2016M590988 and 2016M592868), Fok Ying-Tong Education Foundation (Grant No. 161107), Natural Science Foundation of Xinjiang Uyghur Autonomous Region (Grant No. 2017D01A35 and 2018D01A12) and Prior Period Project of Xinjiang Agricultural University (Grant No. XJAU201505 and XJAU201506).

Compliance with ethical standards

Conflict of interest

None.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

References

  1. Becher P, Tautz N (2011) RNA recombination in pestiviruses: cellular RNA sequences in viral genomes highlight the role of host factors for viral persistence and lethal disease. RNA Biol 8:216–224CrossRefGoogle Scholar
  2. Burns AJ, Herbert TM, Ward SM, Sanders KM (1997) Interstitial cells of Cajal in the Guinea-pig gastrointestinal tract as revealed by c-kit immunohistochemistry. Cell Tissue Res 290:11–20CrossRefGoogle Scholar
  3. Chen MK, Liu SZ, Zhang L (2012) Immunoinflammation and functional gastrointestinal disorders. Saudi J Gastroenterol 18:225–229.  https://doi.org/10.4103/1319-3767.98420 CrossRefGoogle Scholar
  4. Dave M et al (2015) Stem cells for murine interstitial cells of cajal suppress cellular immunity and colitis via prostaglandin E2 secretion. Gastroenterology 148:978–990CrossRefGoogle Scholar
  5. Deng M et al (2015) Prevalence study and genetic typing of bovine viral diarrhea virus (BVDV) in four bovine species in China. PLoS One 10:e0121718CrossRefGoogle Scholar
  6. Farrugia G (2008) Interstitial cells of Cajal in health and disease. Neurogastroenterol Motil 20(Suppl 1):54–63.  https://doi.org/10.1111/j.1365-2982.2008.01109.x CrossRefGoogle Scholar
  7. Fu Q et al (2015) Lentivirus-mediated Bos taurus bta-miR-29b overexpression interferes with bovine viral diarrhoea virus replication and viral infection-related autophagy by directly targeting ATG14 and ATG9A in Madin-Darby bovine kidney cells. J Gen Virol 96:85–94CrossRefGoogle Scholar
  8. Gao J et al (2013) Seroprevalence of bovine viral diarrhea infection in yaks (Bos grunniens) on the Qinghai-Tibetan plateau of China. Trop Anim Health Prod 45:791–793CrossRefGoogle Scholar
  9. Jang DE et al (2018) Neuronal nitric oxide synthase is a novel biomarker for the interstitial cells of Cajal in stress-induced diarrhea-dominant irritable bowel syndrome. Dig Dis Sci 63:619–627CrossRefGoogle Scholar
  10. Kaji N et al (2016) Nitric oxide-induced oxidative stress impairs pacemaker function of murine interstitial cells of Cajal during inflammation. Pharmacol Res 111:838–848CrossRefGoogle Scholar
  11. Klein S et al (2013) Interstitial cells of Cajal integrate excitatory and inhibitory neurotransmission with intestinal slow-wave activity. Nat Commun 4:1630.  https://doi.org/10.1038/ncomms2626 CrossRefGoogle Scholar
  12. Kuca T et al. (2018) Identification of Conserved Amino Acid Substitutions During Serial Infection of Pregnant Cattle and Sheep With Bovine Viral Diarrhea Virus. Frontiers in microbiology 9Google Scholar
  13. Lanyon SR, Hill FI, Reichel MP, Brownlie J (2014) Bovine viral diarrhoea: pathogenesis and diagnosis. Vet J 199:201–209.  https://doi.org/10.1016/j.tvjl.2013.07.024 CrossRefGoogle Scholar
  14. Laureyns J, Ribbens S, de Kruif A (2010) Control of bovine virus diarrhoea at the herd level: reducing the risk of false negatives in the detection of persistently infected cattle. Vet J 184:21–26CrossRefGoogle Scholar
  15. Liang X, Ji X, Ran D (2016) Isolation and identification of bovine viral diarrhea virus TC strain and sequence analysis of E0 gene. Animal Husbandry & Veterinary Medicine 48:36–40Google Scholar
  16. Lu G, Qian X, Berezin I, Telford G, Huizinga J, Sarna S (1997) Inflammation modulates in vitro colonic myoelectric and contractile activity and interstitial cells of Cajal. Am J Physiol Gastrointest Liver Physiol 273:G1233–G1245CrossRefGoogle Scholar
  17. Maeda H, Yamagata A, Nishikawa S, Yoshinaga K, Kobayashi S, Nishi K, Nishikawa S (1992) Requirement of c-kit for development of intestinal pacemaker system. Development 116:369–375Google Scholar
  18. Mikkelsen HB (2010) Interstitial cells of Cajal, macrophages and mast cells in the gut musculature: morphology, distribution, spatial and possible functional interactions. J Cell Mol Med 14:818–832.  https://doi.org/10.1111/j.1582-4934.2010.01025.x CrossRefGoogle Scholar
  19. Mostafa RM, Moustafa YM, Hamdy H (2010) Interstitial cells of Cajal, the maestro in health and disease. World J Gastroenterol 16:3239–3248CrossRefGoogle Scholar
  20. Pinior B et al (2017) A systematic review of financial and economic assessments of bovine viral diarrhea virus (BVDV) prevention and mitigation activities worldwide. Prev Vet Med 137:77–92.  https://doi.org/10.1016/j.prevetmed.2016.12.014 CrossRefGoogle Scholar
  21. Radenkovic G, Radenkovic D, Velickov A (2018) Development of interstitial cells of Cajal in the human digestive tract as the result of reciprocal induction of mesenchymal and neural crest cells. J Cell Mol Med 22:778–785.  https://doi.org/10.1111/jcmm.13375 Google Scholar
  22. Reed LJ, Muench H (1938) A simple method of estimating fifty per cent endpoints. Am J Epidemiol 27:493–497CrossRefGoogle Scholar
  23. Sanders KM, Ward SM, Koh SD (2014) Interstitial cells: regulators of smooth muscle function. Physiol Rev 94:859–907.  https://doi.org/10.1152/physrev.00037.2013 CrossRefGoogle Scholar
  24. Sang DK, Sanders KM, Ward SM (2010) Spontaneous electrical rhythmicity in cultured interstitial cells of Cajal from the murine small intestine. J Physiol 513:203–213Google Scholar
  25. Streutker CJ, Huizinga JD, Driman DK, Riddell RH (2007a) Interstitial cells of Cajal in health and disease. Part I: normal ICC structure and function with associated motility disorders. Histopathology 50:176–189.  https://doi.org/10.1111/j.1365-2559.2006.02493.x CrossRefGoogle Scholar
  26. Streutker CJ, Huizinga JD, Driman DK, Riddell RH (2007b) Interstitial cells of Cajal in health and disease. Part II: ICC and gastrointestinal stromal tumours. Histopathology 50:190–202.  https://doi.org/10.1111/j.1365-2559.2006.02497.x CrossRefGoogle Scholar
  27. Thomsen L, Robinson TL, Lee JC, Farraway LA, Hughes MJ, Andrews DW, Huizinga JD (1998) Interstitial cells of Cajal generate a rhythmic pacemaker current. Nat Med 4:848–851CrossRefGoogle Scholar
  28. Thuneberg L (1999) One hundred years of interstitial cells of Cajal. Microsc Res Tech 47:223–238.  https://doi.org/10.1002/(SICI)1097-0029(19991115)47:4<223::AID-JEMT2>3.0.CO;2-C CrossRefGoogle Scholar
  29. Weiss M, Hertig C, Strasser M, Vogt H, Peterhans E (1994) Bovine virus diarrhea/mucosal disease: a review. Schweiz Arch Tierheilkd 136:173–185Google Scholar
  30. Woodhouse SD et al (2010) Transcriptome sequencing, microarray, and proteomic analyses reveal cellular and metabolic impact of hepatitis C virus infection in vitro. Hepatology 52:443–453CrossRefGoogle Scholar
  31. Yang D-S et al (2007) Serological investigation of bovine viral diarrhea in Fujian Province in 2006 [J]. Progress in Veterinary Medicine 9:012Google Scholar
  32. Zhang L, Zhao B, Liu W, Ma R, Wu R, Gao Y (2017) Cotransplantation of neuroepithelial stem cells with interstitial cells of Cajal improves neuronal differentiation in a rat aganglionic model. J Pediatr Surg 52:1188–1195CrossRefGoogle Scholar
  33. Zhou J, O’Connor MD, Ho V (2017) The potential for gut organoid derived interstitial cells of Cajal in replacement therapy. Int J Mol Sci 18:2059CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Shengnan Li
    • 1
  • Xinyan Hu
    • 1
  • Ruixin Tian
    • 1
  • Yanting Guo
    • 1
  • Junzhen Chen
    • 1
  • Zhen Li
    • 1
  • Xinyan Zhao
    • 1
  • Ling Kuang
    • 1
  • Duoliang Ran
    • 1
  • Hongqiong Zhao
    • 1
  • Xiaohong Zhang
    • 1
  • Jinquan Wang
    • 1
  • Lining Xia
    • 1
  • Jianbo Yue
    • 2
  • Gang Yao
    • 1
    Email author
  • Qiang Fu
    • 1
    Email author
  • Huijun Shi
    • 1
    Email author
  1. 1.College of Veterinary MedicineXinjiang Agricultural UniversityUrumqiChina
  2. 2.Department of Biomedical SciencesCity University of Hong KongHong KongChina

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