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Systemic LPS induces toll-like receptor 3 (TLR3) expression and apoptosis in testicular mouse tissue

  • Lene N. Nejsum
  • Adrian Piec
  • Monika Fijak
  • Christina V. Ernstsen
  • Dania Fischer
  • Thorsten J. Maier
  • Ralf Kinscherf
  • Rainer Hofmann
  • Anja UrbschatEmail author
Regular Article
  • 45 Downloads

Abstract

It is well known that sepsis and inflammation reduce male fertility. Within the testis, toll-like receptor 3 (TLR3) is constitutively expressed and recognizes double-stranded RNA (dsRNA) from viruses, degraded bacteria, damaged tissues and necrotic cells. To characterize the potential role of TLR3 in response to testicular infections, its expression and downstream signaling were investigated upon challenge with lipopolysaccharides (LPS) in two mouse strains that differ in their immuno-competence regarding T cell-regulated immunity. Thereto, Balb/c and Foxn1nu mice were randomized into six interventional groups treated with either i.v. application of saline or LPS followed by 20 min, 5 h 30 min and 18 h of observation and two sham-treated control groups. LPS administration induced a significant stress response; the amplification was manifested for TLR3 and interleukin 6 (IL6) mRNA in the impaired testis 5 h 30 min after LPS injection. TLR3 immunostaining revealed that TLR3 was primarily localized in spermatocytes. The TLR3 expression displayed different temporal dynamics between both mouse strains. However, immunofluorescence staining indicated only punctual interferon regulatory factor 3 (IRF3) expression upon LPS treatment along with minor alterations in interferon β (IFNβ) mRNA expression. Induction of acute inflammation was closely followed by a significant shift of the Bax/Bcl2 ratio to pro-apoptotic signaling accompanied by augmented TUNEL-positive cells 18 h after LPS injection with again differing patterns in both mouse strains. In conclusion, this study shows the involvement of TLR3 in response to LPS-induced testicular inflammation in immuno-competent and -incompetent mice, yet lacking transmission into its signaling pathway.

Keywords

TLR3 Testis Sepsis Lipopolysaccharide Apoptosis 

Notes

Acknowledgements

We especially thank Ecatarina Oplesch, Michael Dreher and Hanne Sidelmann for their technical support.

Funding

This study was funded by a junior researchers grant by the Goethe University Frankfurt (Fokus Programme, Förderlinie A) to AU.

Compliance with ethical standards

Ethical approval

All procedures involving animals were approved by the Animal Care and Use Committee of the state of Hesse in Germany (V54-19 c 20/15-FK/1037). Surgery and animal care was performed in accordance with the “Guide for the care and use of laboratory animals” (National Institutes of Health, volume 25, no. 28, revised 1996), EU Directive 86/609 EEC and German Protection of Animals Act.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

441_2019_3022_Fig7_ESM.png (453 kb)
Supplemental Fig. 1

Applying the same settings for both mouse strains Foxn1nu mice exhibited a higher constitutive immuno-histochemical expression of TLR3 (images from three animals 18 h following NaCl injection) (a - b”). Positive and negative staining control for TLR3 in order to prove the specificity of TLR3 expression using sections from a murine (C57BL/6) kidney 24 h after ischemia and reperfusion injury (IR) and a control kidney (c, c’). (PNG 453 kb)

441_2019_3022_MOESM1_ESM.tif (22.6 mb)
High Resolution Image (TIF 23115 kb)

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Copyright information

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

Authors and Affiliations

  1. 1.Department of Clinical MedicineAarhus UniversityAarhusDenmark
  2. 2.Clinic of Urology and Pediatric UrologyPhilipps-University MarburgFrankfurtGermany
  3. 3.Department of Anatomy and Cell BiologyJustus-Liebig University GießenGiessenGermany
  4. 4.Department of Anesthesiology, Intensive Care Medicine and Pain Therapy, University Hospital FrankfurtGoethe-University Frankfurt/MainFrankfurtGermany
  5. 5.Department of BiomedicineAarhus UniversityAarhus CDenmark
  6. 6.Department of Medical Cell Biology, Institute for Anatomy and Cell BiologyPhilipps-University of MarburgFrankfurtGermany

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