Advertisement

Identification and characterization of an antimicrobial peptide, lysozyme, from Suncus murinus

  • Shota Takemi
  • Shiomi Ojima
  • Toru Tanaka
  • Takafumi Sakai
  • Ichiro SakataEmail author
Regular Article

Abstract

Lysozyme is one of the most prominent antimicrobial peptides and has been identified from many mammalian species. However, this enzyme has not been studied in the order Insectivora, which includes the most primitive placental mammals. Here, we done the lysozyme cDNA from Suncus murinus (referred to as suncus, its laboratory name) and compare the predicted amino acid sequence to those from other mammalian species. Quantitative PCR analysis revealed a relatively higher expression of this gene in the spleen and gastrointestinal tract of suncus. The lysozyme-immunopositive (ip) cells were found mainly in the red pulp of the spleen and in the mucosa of the whole small intestine, including the follicle-associated epithelium and subepithelial dome of Peyer’s patches. The lysozyme-ip cells in the small intestine were mostly distributed in the intestinal crypt, although lysozyme-expressing cells were found not only in the crypt but also in the villi. On the other hand, only a few lysozyme-ip cells were found in the villi and some granules showing intense fluorescence were located toward the lumen. As reported for other mammals, Ki67-ip cells were localized in the crypt and did not co-localize with the lysozyme-ip cells. Moreover, fasting induced a decrease in the mRNA levels of lysozyme in the intestine of suncus. In conclusion, we firstly identified the lysozyme mRNA sequence, clarified expression profile of lysozyme transcripts in suncus and found a unique distribution of lysozyme-producing cells in the suncus intestine.

Keywords

Lysozyme Suncus Intestine Spleen Peyer’s patch 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

441_2019_2991_MOESM1_ESM.pdf (196 kb)
ESM 1 (PDF 195 kb)

References

  1. Ageitos JM, Sanchez-Perez A, Calo-Mata P, Villa TG (2017) Antimicrobial peptides (AMPs): ancient compounds that represent novel weapons in the fight against bacteria. Biochem Pharmacol 133:117–138CrossRefGoogle Scholar
  2. Allison AFVD (1922) Observations on a bacteriolytic substance (“lysozyme”) found in secretions and tissues. Br J Exp Pathol 3:252–260Google Scholar
  3. Bel S, Pendse M, Wang Y, Li Y, Ruhn KA, Hassell B, Leal T, Winter SE, Xavier RJ, Hooper LV (2017) Paneth cells secrete lysozyme via secretory autophagy during bacterial infection of the intestine. Science 357:1047–1052Google Scholar
  4. Callewaert L, Michiels CW (2010) Lysozymes in the animal kingdom. J Biosci 35:127–160CrossRefGoogle Scholar
  5. Cramer EM, Breton-Gorius J (1987) Ultrastructural localization of lysozyme in human neutrophils by immunogold. J Leukoc Biol 41:242–247CrossRefGoogle Scholar
  6. Cross M, Mangelsdorf I, Wedel A, Renkawitz R (1988) Mouse lysozyme M gene: isolation, characterization, and expression studies. Proc Natl Acad Sci U S A 85:6232–6236CrossRefGoogle Scholar
  7. Gomez JM, Verdu M, Gonzalez-Megias A, Mendez M (2016) The phylogenetic roots of human lethal violence. Nature 538:233–237CrossRefGoogle Scholar
  8. Gordon S, Todd J, Cohn ZA (1974) In vitro synthesis and secretion of lysozyme by mononuclear phagocytes. J Exp Med 139:1228–1248CrossRefGoogle Scholar
  9. Hodin CM, Lenaerts K, Grootjans J, de Haan JJ, Hadfoune M, Verheyen FK, Kiyama H, Heineman E, Buurman WA (2011) Starvation compromises Paneth cells. Am J Pathol 179:2885–2893CrossRefGoogle Scholar
  10. Horn CC, Kimball BA, Wang H, Kaus J, Dienel S, Nagy A, Gathright GR, Yates BJ, Andrews PL (2013) Why can't rodents vomit? A comparative behavioral, anatomical, and physiological study. PLoS One 8:e60537CrossRefGoogle Scholar
  11. Ito H, Nishibayashi M, Kawabata K, Maeda S, Seki M, Ebukuro S (2003) Induction of Fos protein in neurons in the medulla oblongata after motion- and X-irradiation-induced emesis in musk shrews (Suncus murinus). Auton Neurosci 107:1–8CrossRefGoogle Scholar
  12. Keshav S, Chung P, Milon G, Gordon S (1991) Lysozyme is an inducible marker of macrophage activation in murine tissues as demonstrated by in situ hybridization. J Exp Med 174:1049–1058CrossRefGoogle Scholar
  13. Klockars M, Osserman EF (1974) Localization of lysozyme in normal rat tissues by an immunoperoxidase method. J Histochem Cytochem 22:139–146CrossRefGoogle Scholar
  14. Klockars M, Reitamo S (1975) Tissue distribution of lysozyme in man. J Histochem Cytochem 23:932–940CrossRefGoogle Scholar
  15. Lelouard H, Henri S, De Bovis B, Mugnier B, Chollat-Namy A, Malissen B, Meresse S, Gorvel JP (2010) Pathogenic bacteria and dead cells are internalized by a unique subset of Peyer's patch dendritic cells that express lysozyme. Gastroenterology 138:173–184 e171–173CrossRefGoogle Scholar
  16. Lewin K (1969) Histochemical observations on Paneth cells. J Anat 105:171–176Google Scholar
  17. Longo VD, Mattson MP (2014) Fasting: molecular mechanisms and clinical applications. Cell Metab 19:181–192CrossRefGoogle Scholar
  18. Murphy WJ, Pringle TH, Crider TA, Springer MS, Miller W (2007) Using genomic data to unravel the root of the placental mammal phylogeny. Genome Res 17:413–421CrossRefGoogle Scholar
  19. Potten CS, Gandara R, Mahida YR, Loeffler M, Wright NA (2009) The stem cells of small intestinal crypts: where are they? Cell Prolif 42:731–750CrossRefGoogle Scholar
  20. Ragland SA, Criss AK (2017) From bacterial killing to immune modulation: recent insights into the functions of lysozyme. PLoS Pathog 13:e1006512CrossRefGoogle Scholar
  21. Sanger GJ, Holbrook JD, Andrews PL (2011) The translational value of rodent gastrointestinal functions: a cautionary tale. Trends Pharmacol Sci 32:402–409CrossRefGoogle Scholar
  22. Satoh Y, Ishikawa K, Tanaka H, Oomori Y, Ono K (1988) Immunohistochemical observations of lysozyme in the Paneth cells of specific-pathogen-free and germ-free mice. Acta Histochem 83:185–188CrossRefGoogle Scholar
  23. Shanahan MT, Carroll IM, Grossniklaus E, White A, von Furstenberg RJ, Barner R, Fodor AA, Henning SJ, Sartor RB, Gulati AS (2014) Mouse Paneth cell antimicrobial function is independent of Nod2. Gut 63:903–910CrossRefGoogle Scholar
  24. Sunkara LT, Jiang W, Zhang G (2012) Modulation of antimicrobial host defense peptide gene expression by free fatty acids. PLoS One 7:e49558CrossRefGoogle Scholar
  25. Suzumoto M, Hotomi M, Fujihara K, Tamura S, Kuki K, Tohya K, Kimura M, Yamanaka N (2006) Functions of tonsils in the mucosal immune system of the upper respiratory tract using a novel animal model, Suncus murinus. Acta Otolaryngol 126:1164–1170CrossRefGoogle Scholar
  26. Takehana K, Masty J, Yamaguchi M, Kobayashi A, Yamada O, Kuroda M, Park YS, Iwasa K, Abe M (1998) Fine structural and histochemical study of equine Paneth cells. Anat Histol Embryol 27:125–129CrossRefGoogle Scholar
  27. Takemi S, Sakata I, Apu AS, Tsukahara S, Yahashi S, Katsuura G, Iwashige F, Akune A, Inui A, Sakai T (2016) Molecular cloning of ghrelin and characteristics of ghrelin-producing cells in the gastrointestinal tract of the common marmoset (Callithrix jacchus). Zool Sci 33:497–504CrossRefGoogle Scholar
  28. Tobi M, Maliakkal B, Zitron I, Alousi M, Goo R, Nochomovitz L, Luk G (1992) Adenoma-derived antibody, Adnab-9 recognizes a membrane-bound glycoprotein in colonic tissue and effluent material from patients with colorectal neoplasia. Cancer Lett 67:61–69CrossRefGoogle Scholar
  29. Tsutsui C, Kajihara K, Yanaka T, Sakata I, Itoh Z, Oda S, Sakai T (2009) House musk shrew (Suncus murinus, order: Insectivora) as a new model animal for motilin study. Peptides 30:318–329CrossRefGoogle Scholar
  30. van der Hee B, Loonen LMP, Taverne N, Taverne-Thiele JJ, Smidt H, Wells JM (2018) Optimized procedures for generating an enhanced, near physiological 2D culture system from porcine intestinal organoids. Stem Cell Res 28:165–171CrossRefGoogle Scholar
  31. Vasquez Cachay ME, Gomez EP, Rodriguez Gutierrez JL, Lira Mejia B, Perez NF, Zanuzzi CN, Barbeito C (2014) Paneth cell identification in the small intestine of Guinea pig offsprings (Cavia porcellus). Anat Rec (Hoboken) 297:856–863CrossRefGoogle Scholar
  32. Wang L, Li J, Li J Jr, Li RX, Lv CF, Li S, Mi YL, Zhang CQ (2016) Identification of the Paneth cells in chicken small intestine. Poult Sci 95:1631–1635CrossRefGoogle Scholar
  33. Wehkamp J, Chu H, Shen B, Feathers RW, Kays RJ, Lee SK, Bevins CL (2006) Paneth cell antimicrobial peptides: topographical distribution and quantification in human gastrointestinal tissues. FEBS Lett 580:5344–5350CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Shota Takemi
    • 1
  • Shiomi Ojima
    • 1
  • Toru Tanaka
    • 2
  • Takafumi Sakai
    • 1
    • 3
  • Ichiro Sakata
    • 1
    Email author
  1. 1.Area of Regulatory Biology, Division of Life Science, Graduate School of Science and EngineeringSaitama UniversitySaitamaJapan
  2. 2.Faculty of Pharmaceutical Sciences, Department of Pharmaceutical and Health SciencesJosai UniversitySakadoJapan
  3. 3.Area of Life-NanoBio, Division of Strategy Research, Graduate School of Science and EngineeringSaitama UniversitySaitamaJapan

Personalised recommendations